ArchiveThe Omicron variant of SARS-CoV-2 is a new variant of concern after Alpha, Beta, Gamma and Delta variants. The amino acid mutations in the viral antigens, especially in the receptor binding region (RBD) of spike protein, were significantly more than those of other variants, which lead to the significant increase of infectivity, transmissibility and immune escape of Omicron variant. In addition, those spike mutations impaired the protective effect of vaccination. When compared to the infection of other variants, the latency of Omicron variant infection was significantly shortened, and the pathogenicity decreased markedly, which is in consistence with the fact that the vast majority of infected individuals showed no symptoms or only mild disease. Exacerbations in patients infected by Omicron variant were often associated with the progress of underlying disease. Early detection and medical isolation of infected persons, careful personal protection measures to cut off transmission routes, and active vaccination to protect susceptible people are key measures to prevent the spread of Omicron variant epidemic. A small number of patients infected with Omicron variant may develop so-called long COVID-19, post-COVID-19 syndrome, or post-COVID-19 condition, which means that long-term follow-up is needed in those patients. Effective anti-Omicron variant therapy can shorten the course of infection, promote the recovery from infection, and also contribute to the control of infection. Therefore, the development of antiviral drugs with ideal cost-benefit ratio and convenient administration is one of the research hotspot in the future.
Objective To analyze the mental health status of medical staff in the Fourth Branch of National Convention and Exhibition Center Makeshift Hospital during the COVID-19 epidemic in Shanghai to lay a theoretical foundation for the mental health and psychological intervention of medical staff in COVID-19 and other public health emergencies. Methods An online questionnaire survey was conducted with the generalized anxiety disorder scale (GAD-7), patient health questionnaire (PHQ-9),and Athens insomnia scale (AIS) before medical staff entering the makeshift hospital and one month later. Results The detection rates of anxiety, depression and insomnia were 18.4%, 22.1% and 27.0% respectively before entering the makeshift hospital, and 28.8%, 59.3% and 64.2% respectively during the follow-up period one month later. The GAD-7, PHQ-9 and AIS scores of medical staff after working in the makeshift hospital for one month increased significantly compared with those at the baseline period(P<0.01). Female and previous history of using sedative and hypnotic drugs were risk factors for increased depression level among medical staff in the makeshift hospital. Conclusions The anxiety, depression and insomnia levels of the medical staff in Shanghai increased after working in the makeshift hospital for one month. It is of great significance for the front-line support work to identify the medical staff with serious psychological problems and carry out psychological intervention in the early stage.
Objective To analyze the potential factors influencing the viral shedding time (duration of nucleic acid positivity) in elderly patients with mild and asymptomatic infection. Methods The clinical data of 1141 elderly (≥60 years)patients with mild and asymptomatic Omicron infection who were admitted to National Exhibition and Convention Center(Shanghai) Cabin Hospital from April 14, 2022 to May 1, 2022 were retrospectively collected, viral shedding time of patients were compared between different groups (age, gender, number of vaccination, hypertension, diabetes). Pearson analysis was adopted to analyze the relationship between age and viral shedding time. Kaplan-Meier curve and Log-rank test were used to evaluate the viral shedding time in elderly patients with different clinical characteristics. Multivariate Cox proportional-hazards regression model was adopted to analyze the factors influencing viral shedding time in elderly patients with Omicron. Results Among 1441 patients,791(54.9%) males and 650(45.1%) females. There were 513(35.6%) patients receiving 0 dose of vaccine, 29(2.0%) patients received 1 dose of vaccine, 405(28.1%) patients received 2 doses of vaccine, 494(34.3%) patients received 3 doses of vaccine. Compared with patients aged 60 to 70 years, patients aged 70 to 80 years had longer viral shedding time (P<0.001). The viral shedding time in patients with hypertension and diabetes was longer than that in patients without hypertension and diabetes (P<0.05). In terms of vaccination, the viral shedding time of patients receiving 2 or 3 doses of vaccine was significantly shorter than that of patients receiving 1 dose of vaccine or none (P<0.05). There was a positive correlation between patient age and viral shedding time, with an R2=0.029 (P<0.001). Kaplan-Meier curve showed that there existed significant difference in viral shedding time between the patients with different vaccination doses (P<0.001), and patients with age ≥70, hypertension and diabetes were all associated with prolonged viral shedding time (P<0.05). Cox regression analysis showed that the age ≥70 years was a risk factor for prolonged viral shedding time, and 2 or 3 doses of vaccine was a protective factor for prolonged viral shedding time (P<0.05). Conclusions Among the elderly population, the viral shedding time would gradually increase with age. Patients who received ≥2 doses of vaccine would have reduced viral shedding time compared with those who received <2 doses of vaccine.
The coronavirus disease 2019 (COVID-19) pandemic is a serious threat to human life, health and social development. In response to this public health event, various COVID-19 vaccines have been rapidly developed around the world. To date, 41 vaccines have been approved for emergency use, and the use of vaccines has significantly reduced the morbidity and mortality of COVID-19. However, with the continuous mutants, especially the emergence of the Omicron variant, challenges to vaccine-induced immune protection are appearing, there is still a long way to go for the continued development of COVID-19 vaccines. This article briefly reviews the research progress of COVID-19 vaccine and its effect on mutant virus strains.
Objective To observe the effects of free sound field blast shock waves on damage effect and neuro behavior in rats, so as to provide the experimental basis for evaluating the biological damage effect of ammunition and proposing the related protective and treatment measures. Methods Ninety SD rats were randomly divided into trinitrotoluene (TNT) explosion group(n=40), thermobaric (TB) explosion group (n=40) and control group (n=10) according to ammunition type and laying distance(18, 20, 25, 30 m). TNT and TB ammunition were detonated in the free field and the shock wave parameters at different distances from the explosion source were measured by pressure transducers. The degree index of injury of rats was evaluated by grass anatomy, histopathology and adjusted severity of injury index (ASII) scoring system. Behavioral changes of the rats were evaluated with Open field test and Morris water maze test. Results The peak pressure and impulse of explosion shock wave decreased continuously with increasing distance from the explosion center in TNT explosion group and TB explosion group. The rats in TNT explosion group died at 18 m (3/10) and 20 m (1/10), while all rats in TB explosion group survived. The dead rats showed a combined lung blast injury. HE staining showed widened alveolar septa and infiltration of red blood cells and inflammatory cells, and no obvious pathological changes were found in brain tissue 24 hours after explosion. Significant correlation existed between ASII score and shock wave peak pressure in TNT explosion groups (r=0.81, P<0.05) and TB explosion groups (r=0.70, P<0.05). The parameters of total move time and total move distance of rats in different positions were shorter in TNT explosion group than in control group, but without statistical significance (P>0.05). Compared with the control group, the move time and distance of rat in 20 m central region were shorter in TNT explosion group (P<0.05). Compared with the control group, the escape latency of rats in TNT explosion group 18 m and TB explosion group 20 m was shortened, the platform crossing time increased, and the residence time in target quadrant prolonged (P<0.05). Conclusions The injury degree of free-field blast shock wave in rats is related to the distance to explosion center with a shock wave peak pressure and impulse dependence. The blast shock wave can affect the behavior and spatial memory ability of rats.
Objective To explore the effects of schisandrin B (Sch B) on myocardial remodeling and electrocardiogram changes of atrial fibrillation rats. Methods A total of 60 SD rats were randomly divided into normal group, model group, positive control group, low-dose group and high-dose group (12 rats each). Except normal group, rats in other groups were injected with Ach-CaCl2 mixed solution through tail vein to establish atrial fibrillation model. From the 4th day of modeling, 1 h before Ach-CaCl2 mixture injection through tail vein injection, rats in positive control group, low-dose and high-dose Sch B group were given intragastric administration of amiodarone 50 mg/kg, Sch 40 mg/kg and Sch 80 mg/kg, respectively, once a day for consecutive 7 days. ECG of each group was recorded during the last caudal vein injection of Ach-CaCl2 mixture. Echocardiography was used to measure left ventricular ejection fraction (LVEF), left ventricular shortening fraction (LVFS), left ventricular end-diastolic diameter (LVEDD)and left ventricular end-systolic diameter (LVESD). The relative expression levels of collagen Ⅰ (Col Ⅰ) and collagen Ⅲ (Col Ⅲ)mRNA were detected by qRT-PCR, the pathological changes of atrial tissue were observed by HE staining, the myocardial fibrosis was detected by Masson staining, and the relative expression levels of p-Akt, p-mTOR and p-S6K were detected by Western blotting. Results The results of echocardiography showed that LVEF and LVFS were higher, while LVEDD and LVESD were shorter in positive control group, low-dose and high-dose Sch B groups than in model group; Compared with the low-dose Sch B group, the LVEF and LVFS increased significantly, and the LVEDD and LVESD shortened significantly in high-dose Sch B group (P<0.05). The results of qRT-PCR showed that the relative expression levels of Col Ⅰ and Col Ⅲ mRNA were lower in positive control group, low-dose and high-dose Sch B group than in model group; Compared with the low-dose Sch B group, the relative expression levels of Col Ⅰ and Col Ⅲ mRNA were decreased in high-dose Sch B group (P<0.05). The results of HE staining showed that the cardiomyocytes in normal group were in order and dense, and the morphology of the cells was normal with no edema in the interstitium. While those in model group were out of order, intercellular space widened and intercellular interstitium was with edema. The injury of myocardial cells in positive control group, low-dose and high-dose Sch B group was significantly improved. Masson staining showed that the percentage of collagen area in atrial tissue of rats decreased in positive control group, low-dose and high-dose Sch B groups than that in model group; Compared with the low-dose Sch B group, the percentage of collagen area in atrial tissue of rats decreased in high-dose Sch B group (P<0.05). Western blotting results showed that the relative protein expression levels of p-Akt, p-mTOR and p-S6K were lower in positive control group, low-dose and high-dose Sch B groups than those in model group;Compared with the low-dose Sch B group, the relative expression levels of p-Akt, p-mTOR and p-S6K proteins were decreased in high-dose Sch B group (P<0.05). Conclusion Sch B can mitigate rats' atrial fibrillation, improve myocardial injury and cardiac dysfunction, possibly by inhibiting the Akt/mTOR/S6K signaling pathway.
Objective To explore the effects and mechanism of long non-coding RNA HLA complex group 18 (HCG18)on human ovarian cancer cell SKOV3 and its cisplatin resistance cell line SKOV3/DDP. Methods RT-PCR was used to detect the expression levels of HCG18 and miR-449 in SKOV3 and SKOV3/DDP cells. Cells were transfected with HCG18 siRNA and treated with DDP (12.5 μg/ml). After 48 h, we evaluated cell viability using the MTT assay, examined cell apoptosis and cell cycle through flow cytometry, and confirmed targeting binding of HCG18, miR-449, and Wnt1 with luciferase reporter assay. Results Compared with the human normal ovarian epithelial cell line NOEC, the HCG18 level was significantly increased (P<0.05), and the miR-449 level was significantly decreased in SKOV3 (P<0.05). Compared with the transfection control group, in the DDP-treated cells (both SKOV3 and SKOV3/DDP), we observed significantly reduced cell viability, enhanced cell apoptosis, and cell cycle G2/M phase arrest (P<0.05). Transfection of MiR-449 inhibitor partly reversed the effects of HCG18 siRNA on cell viability, cell apoptosis, and cell cycle in both SKOV3 and SKOV3/DDP cells (P<0.05). The luciferase reporter results showed that HCG18 could bind to the 3'-UTR of miR-449, and miR-449 could bind to the 3'-UTR of Wnt1. Transfection of HCG18 siRNA significantly increased the levels of miR-449 (P<0.05). Transfection of HCG18 siRNA or miR-449 mimic significantly decreased the mRNA levels of Wnt1(P<0.05). Conclusions HCG18 knockdown could inhibit cell viability, promote cell apoptosis and cell cycle G2/M phase arrest through miR-449/Wnt1 signaling pathway, and thus promote cisplatin sensitivity of human ovarian cancer cells.
Objective To explore the correlation and potential mechanism of plasma magnesium (Mg2+) concentration with blood lipids and uric acid. Methods The physical examination data of healthy population from September 2018 to May 2021 were collected from the Second Affiliated Hospital of Xi'an Jiaotong University, and divided into two groups according to the plasma Mg2+ concentration (low Mg2+ group, ≤1.65 mmol/L; high Mg2+ group, >1.65 mmol/L). The differences of blood lipids and uric acid were compared between the two groups. Spearman correlation analysis was performed to analyze the correlation of plasma Mg2+ concentration and the metabolism of blood lipids and uric acid. Subgroups were set up according to gender and age, and based on the Comparative Toxicogenomics Database (CTD) and other disease-related databases, genes related to Mg2+ and dyslipidemia were extracted and matched, protein interaction (PPI) network was constructed, and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were performed. Results Uric acid [(288.88±80.44) mg/dl vs. (325.00±83.38) mg/dl, P<0.001], total cholesterol [TC, (4.27±0.85) mmol/L vs. (4.52±0.87) mmol/L, P<0.001],triglyceride [TG, (1.31±0.97) mmol/L vs. (1.70±1.33) mmol/L, P<0.001] and low-density lipoprotein cholesterol [LDL-C,(2.62±0.76) mmol/L vs. (2.85±0.75) mmol/L, P<0.001] were significantly increased in healthy population with higher Mg2+ concentration, while high-density lipoprotein cholesterol (HDL-C) decreased significantly [(1.33±0.34) mmol/L vs.(1.25±0.30) mmol/L, P<0.001]. Spearman correlation analysis showed that plasma Mg2+ was positively correlated with uric acid(r=0.237, P<0.001), TC (r=0.154, P<0.001), TG (r=0.254, P<0.001), LDL-C (r=0.170, P<0.001), while negatively correlated with HDL-C (r=–0.154, P<0.001). Analyzed results in male and female subgroups were basically consistent with the above results, and the differences mainly come from the age group of 20-40 years old and 40-60 years old. In addition, there were 12 matched genes between Mg2+ and dyslipidemia. GO and KEGG enrichment analysis indicated that Mg2+ may act on insulin, SREBF1, HMGCR, LCAT, CD36 and other liver lipid synthesis and metabolic targets, and thus affect adenosine monophosphate protein kinase (AMPK) signaling, insulin resistance, and atherosclerosis. Conclusion The raised plasma Mg2+ concentration is often correlated with dyslipidemia and hyperuricemia in healthy population. Magnesium may involve in liver lipid metabolism and insulin pathway, and play a role in dyslipidemia, insulin resistance and atherosclerosis.
Objective To summarize and analyze the clinical features, treatment and prognosis of children with severe anti-N-methyl-D-aspartate receptor (anti-NMDAR) encephalitis. Methods A total of 20 cases clinical data of children patients with severe anti-NMDAR encephalitis were collected from the Department of Internal Neurology and Pediatric ICU of Children's Hospital affiliated to Chongqing Medical University from January 2014 to September 2021. The clinical features, treatment and prognosis were analyzed. Modified Rankin scale (mRS) was used to evaluate the short-term prognosis of the patients, and divided them into favourable prognosis group (mRS <2) and unfavourable prognosis group (mRS ≥2, or death), and analyze the influencing factor to prognosis. Results A total of 20 children with severe anti-NMDAR encephalitis were included, among them 11 girls and 9 boys with on-set age of (9.2±3.7) years. convulsion attack and mental behaviour disorder were the first neurological symptoms, and the main complications included consciousness disturbance (85.0%), status epilepticus (60.0%), central hypoventilation (40.0%),acute intracranial hypertension (35.0%), multiple organ dysfunction (20.0%), shock (20.0%) and rhabdomyolysis (15.0%), while no multiple organ dysfunction, shock and rhabdomyolysis occurred in favourable prognosis group. Moreover, slow wave activity was critical feature of first electroencephalography (EEG) abnormalities (95.0%), and none of the patients in favourable prognosis group had epileptiform waves and δ-brushes. The first-line immune treatment was received by all the children patients including the combination therapies of methyllprednisolone, intravenous immunoglobulin (IVIG) or plasma exchange (TPE), among them 2 cases received the second-line immunotherapy with rituximab. Glasgow coma score (GCS) scores improved after immunotherapy compared with pre-treatment. Short-term follow-up was performed for 3 months. Finally, 4 of the children patients died and 16 survived with neurological deficits, mainly including cognitive impairment (12 cases, 75.0%), language impairment (10 cases,62.5%) and motor deficits (8 cases, 50.0%), average mRS score was 2.81±1.55. The time from admission to performed the antibody testing was significantly shorter in favourable prognosis group than that in the unfavourable prognosis group. Conclusions For children patients with severe anti-NMDAR encephalitis, in addition to early testing the cerebrospinal fluid antibody, completing electroencephalogram and early initiation of immunotherapy, attention should also be paid to the management of central respiratory, cardiovascular and status epilepticus. Immunotherapy is effective in majority of the patients. However, children with severe anti-NMDAR encephalitis have a poor short-term prognosis.
Objective To explore the clinical value of dual-layer spectral detector CT (DLCT) in distinguishing diagnosis of pulmonary primary malignant tumor, chronic inflammation and tuberculosis by measuring and analyzing the parameters and conventional CT signs. Methods The clinical data of 345 patients with pulmonary lesions were collected from August 2020 to June 2021, who underwent DLCT chest enhanced scan and obtained pathological results in People's Hospital of Gansu Province, and then divided into three groups: pulmonary primary malignant tumor group (n=187), chronic inflammation group (n=101) and tuberculosis group (n=57). The conventional CT signs of the three groups were retrospectively analyzed and the DLCT parameters were measured. The logistic regression analysis was performed for parameters with statistically significant differences, and then conventional CT signs diagnostic model, DLCT parameter diagnostic model and combined diagnostic model were established. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic efficacy of each model. Delong test was used to compare the AUC of each models. Results In distinguishing the conventional CT signs of the three lesions, statistical differences existed in the following indicators: the distance of lesions to pleura (P=0.009), morphology (P<0.001), density (P=0.001), the boundary between lesions and lung (P=0.001), lobulation (P<0.001), liquefaction necrosis (P=0.003), vascular cluster sign(P<0.001), halo sign (P=0.003), satellite focus (P=0.045), pleural effusion (P=0.002), enlarged lymph nodes in the mediastinum(P<0.001), effective atomic number (Zeff), iodine concentration (IC), normalization iodine concentration (NIC), energy spectrum curve slope (λHU), and arterial enhancement fraction (AEF) (P<0.001) both in the arterial phase (AP) and venous phase (VP).In the differential diagnosis of pulmonary primary malignant tumor and chronic inflammation, the boundary between lesion and lung tissue (P=0.009), lobulation (P<0.001), liquefaction necrosis (P<0.001), halo sign (P=0.025), mediastinal lymphadenopathy(P<0.001), λHU-AP (P=0.037) and λHU-VP (P=0.029) are independent influencing factors. In the differential diagnosis of pulmonary primary malignant tumor and tuberculosis, lesion morphology (P=0.019), vascular cluster sign (P=0.009), satellite focus (P=0.006),pleural effusion (P=0.001), AEF (P=0.041), λHU-AP (P=0.038) and λHU-VP (P<0.001) are independent influencing factors. Pleural effusion (P=0.002), mediastinal lymphadenopathy (P<0.001), NIC-VP (P=0.001), Zeff-VP (P=0.043), λHU-AP (P=0.015) and λHU-VP (P=0.023) are independent influencing factors in the differential diagnosis of chronic inflammation and tuberculosis. To pathology results for the gold standard, the AUC of conventional CT signs diagnostic model for distinguishing pulmonary primary malignant tumor and chronic inflammation, pulmonary primary malignant tumor and tuberculosis, chronic inflammation and tuberculosis were 0.827, 0.770 and 0.753. The AUC of DLCT parameter values for distinguishing pulmonary primary malignant tumor, chronic inflammation and tuberculosis were 0.905 0.909 and 0.824. The AUC of the combined model for distinguishing pulmonary primary malignant tumor, chronic inflammation and tuberculosis were 0.929, 0.942 and 0.889. Conclusion DLCT parameters combined with conventional CT signs may improve the differential diagnosis efficiency of pulmonary primary malignant tumor, chronic inflammation and tuberculosis.
Objective To report the clinicopathological features and treatment of 5 cases of primary ovarian strumal carcinoid and review the literature for improving the understanding to this disease. Methods The clinical data of 5 cases of primary ovarian strumal carcinoid collected in the Affiliated Jinhua Hospital, Zhejiang University School of Medicine (3 cases) and Dongyang People's Hospital (2 cases) from January 2009 to December 2020, and retrospectively analyzed by clinical features, microscopic appearance and immunohistochemical stains. The clinicopathological features and treatment methods of primary ovarian strumal carcinoid were summarized based on combined literature reports and retrieval of relevant literature databases of 28 patients. Results Case 1 and case 5 were primary ovarian strumal carcinoid, case 2 and case 4 were primary ovarian strumal carcinoid mixed with cystic teratoma. In addition, mucinous carcinoid was also seen in case 2. Case 3 was primary ovarian strumal carcinoid mixed with mucinous carcinoid. Immunohistochemical found the thyroid follicles of all the 5 cases were positive for thyroid markers, the carcinoid components were positive for neuroendocrine markers and somatostatin receptor 2 (SSTR2). Mucinous carcinoid of both case 2 and case 3 were positive for synaptophysin (Syn), chromogranin A (CgA) and caudal type homeobox transcription factor 2(CDX2). The Ki-67 proliferation index of all the 5 cases were less than 2%. In addition, the thyroid follicles of 5 cases also showed different levels of expression for neuroendocrine markers. During follow-up, 4 cases showed no recurrence or metastasis, and one case lost. A total of 28 cases of primary ovarian strumal carcinoid were searched in PubMed from 1971 to 2021 and SEER database from 1988 to 2011. The mean age of the patients was 51 and 46 years old. Except for one case of lymph node metastasis and one case of death, no tumor recurrence and metastasis were found during follow-up. Conclusions Primary ovarian strumal carcinoid is a kind of rare ovarian tumor. Some cases can be mixed with cystic teratoma and mucinous carcinoid. The patients often show no typical clinical symptoms. Missed diagnosis should be avoided during pathological sampling and diagnosis. Surgical methods that retain more organ functions are recommended in stage Ⅰ patients. The prognosis is generally excellent but long-term follow-up is needed to exclude recurrence and occult metastasis.
In recent years, benefiting from the increasing understanding of tumor immunity, the application of immunotherapy especially immune checkpoint inhibitors in the clinic has achieved great success. Although immunotherapy shows great potential for antitumor therapy, many challenges are currently facing. The limited benefit population is one of the important issues, and the development of new therapeutic strategies to increase the proportion of the population benefiting immunotherapy is of great clinical importance. As one of the traditional means of antitumor therapy, increasing evidence shows that it plays an important role in antitumor immune mechanisms. Chemotherapy can improve the sensitivity of tumor cells to programmed death-1(PD-1)/PD-L1 inhibitors by affecting the immune microenvironment and activating antitumor immune responses. Moreover, chemotherapy can affect the tumor cells themselves, upregulate the expression of PD-L1, and induce PD-L1-mediated tumor immunosuppressive phenotype, thereby increasing the therapeutic targets of PD-1/PD-L1 inhibitors. A large body of evidence from clinical studies has also suggested a synergistic effect between combination chemotherapy and immune checkpoint inhibitors in multiple malignancies, and combination therapy is expected to be a novel immunotherapeutic strategy. This paper systematically reviews the evidence from relevant basic and clinical studies, analyzes and summarizes the interaction relationship between chemotherapy and immunotherapy, aiming to provide new ideas for enhancing antitumor immune responses, and at the same time provides a corresponding theoretical basis for clinical combination chemotherapy and immunotherapy.
Growth hormone (GH) and insulin-like growth factor 1 (IGF-1) are the important endocrine hormones for human, and their associated diseases (e. g. acromegaly and growth hormone deficiency) can lead to obviously increased morbidity and mortality secondary to cardiovascular, cerebrovascular and respiratory diseases. Clinically, the determination and closely monitoring of serum GH and IGF-1 are important means for diagnosing, assessing curative effect and formulating post-treatment follow-up plans of such diseases, and have received extensive concern from clinicians. Although the levels of serum GH and IGF-1 are consistent in most cases, more and more studies have reported that the results of serum GH and IGF-1 are unreasonable or even contradictory. Therefore, in the conditions of inconsistent serum GH and IGF-1 levels, reliable biochemical criteria for assessing disease status and control degree are currently controversial, and their rational interpretation remains a clinical challenge. In present paper, the influence of serum GH and IGF-1 detection methods, physiological and pathological states, and drugs on the detection results and the possible reasons or mechanisms are explained in detail, and proposes that it is necessary to carefully sort out the factors that may lead to unreasonable results in clinic, in order to judge the detection results more accurately and conduct appropriate diagnosis and treatment of the related diseases.