Fecal microbiota transplantation (FMT) is becoming a new approach for dysbiosis conditions. It means that fecal microbiota from healthy donors could be transplanted into the recipient's digestive tract, to modulate their tissue and organ function and intestinal microecology, thus gaining a therapeutic effects for dysbiosis disorders. The FMT rescue program aims to provide standard FMT technology and products to military institutions for treating antibiotic-associated diarrhea like clostridium difficile infection (CDI) and other dysbiosis conditions with the FMT indication. The program contents include the purpose, the application scope, the process and technology for rescue use, the composition and responsibility of institutions, etc.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is highly infectious and pathogenic.Although it mainly involves the respiratory system, it can also invade the nervous system and lead to a variety of neurological symptoms and diseases, further complicating the clinical conditions of the patients. In order to assist physicians and neurologists in understanding the pathogenesis, clinical features, diagnostic procedures, therapeutic principles, and clinical outcomes of the diseases,the experts of Neurology Branch of Chinese Medical Doctor Association wrote this expert recommendation based on present research articles and clinical practices about the epidemiology, clinical symptoms, diagnostic algorithms, treatment and prognosis of neurological diseases caused by coronavirus disease 2019, in order to provide reference for clinical diagnosis and treatment.

Objective To analyze the genotypic characteristics of resistance-associated mutation in reverse-transcriptase(RT) domain of hepatitis B virus (HBV) in treatment-experienced chronic hepatitis B (CHB) patients with low-level viremia (LLV). Methods CHB patients with LLV, who admitted to the Fifth Medical Center of Chinese PLA General Hospital from September 2007 to August 2019, were retrospectively enrolled. Their serum HBV DNA was extracted, and a nested PCR was used to amplify an HBV RT fragment, 1225 bp in length (nt54-nt1278), containing the HBV RT gene (nt130-nt1161) and S gene (nt155-nt835).PCR products were bidirectionally sequenced, and a molecular evolutionary tree analysis of the RT/S gene sequence was performed with MEGA 4 software, with the Hepacivirus database (http://www.ncbi.nlm.nih.gov/projects/genotyping/formpage.Cgi) with the standard sequence as the reference for HBV genotyping. Results Among the 2096 patients, 82.4% were male, mean age was(45.6±12.0) years, HBV DNA was (2.5±0.5) log₁₀ IU/ml. Lamivudine resistance related mutations accounted for 58.0%, with M204V and L180M+M204V as the main forms; Adefovir dipivoxil resistance associated mutations accounted for 21.2%, with A181V and N36T as the predominant forms; Entecavir resistance associated mutations accounted for 17.9%, with L180M+M204I/V+T184L/A/S/I and L180M+M204V+S202G being the predominant forms; Multidrug resistance accounted for 2.9%, with L180M+M204I/V+A181V and L180M+M204V+A181V+N236T+S202G being the predominant forms. Conclusion Lamivudine and Adefovir resistance are the main drug-resistant mutation genotypes in the treated CHB patients with LLV, which reduces the gene barrier of entecavir resistance and increases the proportion of entecavir resistance. Therefore, the first-line nucleos(t)ide analogues with strong effect and low resistance should be selected for antiviral treatment.

Objective To study the effect of Wuzi Yanzong Pill (WYP) on neurotrophic factors and apoptosis in the brain of mice with Parkinson's disease (PD). Methods MPTP-induced PD model in mice was established to evaluate the therapeutic effect of WYP by the behavioral gait, pole test, and brain pathological TH-positive neurons in the substantia nigra (SN). We used immunofluorescence, RT-qPCR, and Western blotting to evaluate the expression of neurotrophic factors (BDNF, GDNF, and CNTF) and apoptosis-associated genes (Bcl-2, Bax, caspase-3, and caspase-9). Results Compared with the PD model group, WYP could reduce the average swinging time of each paw (P<0.05)and pole test time (P<0.001); The number of TH positive cells (P<0.05)and TH protein expression (P<0.05) increased, which inhibited dopaminergic neuron injury. Compared with the PD model group,WYP increased the expression of BDNF, CNTF, GDNF, and Bcl-2 (P<0.05) and decreased the expression of Bax, caspase-3, and caspase-9 (P<0.05). Conclusion WYP shows therapeutic potential in PD, and its mechanism may be related to the anti-apoptotic effect and increased secretion of neurotrophic factors in SN of the brain in the MPTP-induced PD model.

Mild traumatic brain injury (mTBI) is common in modern warfare. If the wounded cannot receive early and effective diagnosis and treatment, long-term headaches, dizziness, insomnia, memory disorders, anxiety, depression and other symptoms will be easily left behind, which will seriously affect the quality of life and combat effectiveness of service members, and bring a huge burden on families and society. In recent years, the US Department of Defense has conducted a lot of research on the pathophysiological mechanism, examination, treatment and prognosis of mTBI, and formulated some guidelines and norms. This article reviews the progress in diagnosis and treatment of mTBI, facilitating the establishment of a standardized treatment system for mTBI.

Objective To investigate the effect and mechanism of ZCCHC12 on epithelial-mesenchymal transition and invasion of differentiated thyroid cancer cells. Methods A total of 50 patients with differentiated thyroid adenocarcinoma admitted to Hebei General Hospital from May 2017 to December 2018 were selected, and set as differentiated thyroid cancer group. In addition, 50 subjects for healthy examination in the hospital during the same period were selected as the healthy control group.The contents of PKA/cAMP response element binding protein (CREB) and p21 in serum was detected by ELISA. Western blotting was used to detect the relative expressions of CREB and p21 in HUM-CELL-0097, TPC-1 and FTC-133 cell lines. TPC-1 cells and FTC-133 cells were taken and set up blank control group (normally cultured), NC si group (transfected with non-specific siRNA),and ZCCHC12 si group (transfected with ZCCHC12 siRNA), ZCCHC12 si+NC pc group (transfected with ZCCHC12 siRNA followed by the transfection with pcDNA.3.1), ZCCHC12 si+CREB pc group (transfected with ZCCHC12 siRNA followed by the transfection with pcDNA.3.1-CREB), ZCCHC12 si+NC si group (transfected with ZCCHC12 siRNA followed by the transfection with non-specific siRNA), ZCCHC12 si+p21 si group (transfected with ZCCHC12 siRNA followed by the transfection with p21 siRNA). Western blotting was performed to detect the relative expressions of ZCCHC12, CREB, P21, E-cadherin and N-cadherin proteins. Transwell method was used to detect the cell invasion ability. Results Compared with healthy control group, the serum content of CREB increased (P<0.05), and of p21 decreased (P<0.01) in differentiated thyroid cancer group. Compared with HUM-CELL-0097 cells, the content of CREB increased (P<0.01), and of p21 decreased (P<0.01) in TPC-1 and FTC-133 cells. Compared with NC si group, the relative expressions of E-cadherin and p21 protein increased, while the relative protein expressions of CREB and N-cadherin and the number of cell migration decreased (P<0.01) in ZCCHC12 si group. The expression of p21 protein in ZCCHC12 si+CREB pc group was lower than that in ZCCHC12 si+NC pc group (P<0.01), which reversed the promoting effect of interfering ZCCHC12 on p21 protein expression. Compared with ZCCHC12 si+NC si group, the relative expression of E-cadherin protein significantly decreased, while the relative expression of N-cadherin protein and the number of cell migration were significantly increased (P<0.01) in the ZCCHC12 si+p21 si group, which reversed the inhibitory effect of ZCCHC12 interference on the epithelial-mesenchymal transition and invasive capacity in differentiated thyroid cancer cells. Conclusions Interfering with ZCCHC12 can effectively inhibit the epithelial-mesenchymal transition and invasion of differentiated thyroid cancer cells by regulating CREB and p21, providing a certain theoretical basis for the treatment of differentiated thyroid cancer.

Objective To investigate the effect and possible mechanism of the alcohol extract of Alisma orientalis on thyroid function of mother rats and neurological function of offspring with subclinical hypothyroidism (SCH). Methods A total of 85 female rats were selected, of which 15 rats acted as sham operation group and treated with sham operation, the other 70 rats underwent thyroidectomy + subcutaneous injection of levothyroxine (L-T₄) to establish the SCH model. Sixty-two of the 70 rats were successfully modeled, and randomly divided into low-dose Alisma ethanol extract group (15 rats, gavage with alcohol extract of Alisma orientalis 9 g/kg), high-dose Alisma ethanol extract group (16 rats, gavage with alcohol extract of Alisma 36 g/kg), L-T₄ group (16 rats, gavage with 4.5 μg/kg L-T₄), model group (15 rats, gavage with equal volume of normal saline), once a day, the intervention time is from 10th day of pregnancy (E10) to 21st day after birth (P21). The thyroid stimulating hormone (TSH) and thyroxine (T₄) of female rats at E17 were detected by electrochemiluminescence. ELISA was used to detect the nerve growth factor(NGF) and brain-derived neurotrophic factor (BDNF) in the hippocampus of offspring rats at P21; Western blotting was performed to detect the expression levels of tropomyosin receptor kinase A (TrkA), p-TrkA, c-AMP-response element binding protein (CREB)and p-CREB in the hippocampus of offspring rats at P21; The escape latency of pups at P40 was measured by the positioning cruise experiment, and the number of times the pups crossed the platform at P41 was measured by the space exploration experiment. Results At E17 gestational age, there was no significant difference in serum T₄ among the 5 groups of female mice (P>0.05). The serum TSH levels were lower in low- and high-dose Alisma ethanol extract group and L-T₄ group than that in model group, and the serum TSH levels were lower in high-dose Alisma ethanol extract group and L-T₄ group than that in low-dose Alisma ethanol extract group (P<0.05), while no significant difference existed in serum TSH levels between high-dose Alisma ethanol extract group and L-T₄ group (P>0.05). At P21, the NGF, BDNF, p-TrkA and p-CREB of hippocampus were significantly higher in low- and high-dose Alisma ethanol extract group and L-T₄ group than those in model group. The NGF, BDNF, p-TrkA and p-CREB were obviously higher in high-dose Alisma ethanol extract group and the hippocampus of L-T₄ group than those in the low-dose Alisma ethanol extract group (P<0.05). There was no statistically significant difference on NGF and BDNF between the high-dose Alisma ethanol extract group and the hippocampus of L-T₄ group (P>0.05). The escape latency of offspring rats was shorter in low- and high-dose Alisma ethanol extract group and L-T₄ group than that in model group, and the times of offspring crossing the platform were more than that in model group (P<0.05). The escape latency of offspring in the L-T₄ group was shorter than that in low-dose Alisma ethanol extract group, and the times of offspring crossing the platform was more than that in low-dose Alisma ethanol extract group(P<0.05). There was no significant difference in the escape latency and the times of crossing the platform between high-dose Alisma ethanol extract group and L-T₄ group (P>0.05). Conclusions The ethanol extract of Alisma orientalis applied to SCH mother rats can improve the thyroid function of the mother rats, promote the expression of neurological factors of offspring rats, and improve their learning and memory function. The mechanism may be related to the activation of the expression of TrkA signaling pathway.

Objective To explore the effect of miR-378a overexpression in modifying bone marrow mesenchymal stem cells (BMMSCs) combined with collagen sponge (CS) scaffold in repairing femoral defects of rats. Methods The BMMSCs were isolated and cultured, and the cell phenotype was identified by flow cytometry. The BMMSCs were transfected with an overexpressed miR-378a lentivirus and a negative unloaded lentivirus, and the transfected cells were divided into three groups: a BMMSCs transfected with overexpressed miR-378a group (LV-miR-378a group), BMMSCs transfected with negative control lentivirus group (LV-miR-NC group) and an untransfected BMMSCs group (control group). The transfection efficiency was detected by flow cytometry. The composite of transfected cells with CS scaffold was prepared, and the compatibility of the cells with the scaffold was observed by scanning electron microscopy. A femoral defect replantation model was established in SD rats, and 15 SD rats were randomly divided into three groups (n=5): LV-miR-378a+CS group (with the lentivirus complex CS scaffold overexpressing miR-378a implanted), LV-miR-NC+CS group (with the BMMSCs complex CS scaffold transfected with negative non-load lentivirus),and CS group (with the CS scaffold implanted only). At the 8th week after operation, the SD rats were sacrificed by CO₂ inhalation.The femur on the operation side was taken for gross observation and micro-CT scanning reconstruction. Bone mineral density(BMD) and bone volume/total volume (BV/TV) were quantified, and HE, Masson and osteopontin (OPN) immunohistochemical staining were used to observe bone repair. Results The phenotyping results of BMMSCs by flow cytometry showed that the positive expression rate of cell surface antigen CD44 and CD29 were 95.5% and 94.7%, respectively, while the positive expression rate of CD45 and CD34 were 0.8% and 0.7%, respectively. Transfection efficiency detected by flow cytometry: compared with the control group, the transfection efficiency of LV-miR-378a group and LV-miR-NC group increased obviously (P<0.05). The lentivirus transfection efficiency of LV-miR-378a group was consistent with that of LV-miR-NC group with no statistically significant difference (P>0.05). The results of scanning electron microscopy showed that the CS scaffold material had a good three-dimensional cavity structure. After the two groups of cells were co-cultured with the CS scaffold for 7 days, the cells in LV-miR-378a group had a larger spreading area on the surface of scaffold material than those in LV-miR-NC group, and the cells grow in clusters with more pseudopod. The gross observation results showed that the bone defect area in LV-miR-378a+CS group was completely repaired 8 weeks after surgery. The incompletely mineralized new bone could be observed in the bone defect centers of both LV-miR-NC+CS group and CS group, and the rough outline of the defect area could still be seen in CS group. The results of micro-CT three-dimensional reconstruction and quantitative analysis showed that 8 weeks after surgery, the repair effect of the bone defect area in LV-miR-378a+CS group was better than in other two groups, and there was more new bone deposition. The BMD and BV/TV values were significantly higher than those in other two groups (P<0.05). The defective areas in both LV-miR-NC+CS group and CS group were not completely repaired, and less new bone deposition and uneven density in CS group. HE and Masson staining results showed that the stent material in LV-miR-378a+CS group degraded completely 8 weeks after surgery, and more mature trabeculae could be seen, the junction area of new bone and host bone showed good continuity, and the trabeculae were connected with each other and had regular morphology. In LV-miR-NC+CS group, mature trabeculae were relatively few, and the junction area of new bone and host bone was not completely connected. In CS group, the stent materials were not completely degraded, and there were still more trabeculae in remodeling and irregular shape. Immunohistochemical staining showed that the expression rate of OPN in LV-miR-378a+CS group was significantly lower than that in other two groups 8 weeks after surgery, (P<0.05), and the expression rate of OPN in LV-miR-NC+CS group was lower than that in CS group (P<0.05). Conclusion Over-expression of miR-378a modified BMMSCs combined with CS scaffold can promote new bone formation.

Intestinal microorganisms are closely related to the pathogenesis of depression, but their specific mechanism of action has not been fully explained. Short-chain fatty acids (SCFA) are metabolites produced by the fermentation of plant polysaccharides, such as dietary fiber and resistant starch mediated by gut microorganisms, which are mainly composed of acetic acid, propionic acid, and butyric acid, and play an important role in the microbiota-intestine-brain axis. Recent studies have found that SCFA can not only regulate intestinal energy metabolism, but also improve depression through blood-brain barrier, immune pathway, endocrine pathway and vagus nerve. SCFA is the focus of scientific research nowadays, and there are few reports on SCFA intervention in depression in China. This article reviews the research progress of SCFA on depression, providing new ideas for clinical research.

Objective To analyze the detection characteristics of novel hepatitis B virus (HBV) serum biomarkers including HBV RNA, hepatitis B core-related antigen (HBcrAg), hepatitis B core antibody (anti-HBc), intrahepatic HBV total DNA (tDNA), and covalently closed circle (cccDNA) in patients with different types of HBV infection. Methods A total of 227 HBV-infected patients, who were hospitalized in the Fifth Medical Center of Chinese PLA General Hospital from January 2017 to December 2019 and underwent liver biopsy, were included in this study. The clinical data were retrospectively analyzed and serum HBV RNA, HBcrAg, anti-HBc, intrahepatic HBV tDNA and cccDNA were quantitatively detected. The characteristics and anti-HBV treatment impact on the above virological biomarkers in patients with different types of clinical illness were analyzed. Results Among 16 patients with acute hepatitis B (AHB), 4 were positive for serum HBV RNA (25.0%), 15 were positive for serum HBcrAg(93.8%), 14 were positive for HBV tDNA (87.5%), and 13 were positive for cccDNA (81.3%) in liver tissues. Among 184 patients with chronic hepatitis B (CHB), 34 were treated with nucleos(t)ide analogues (NAs), and their HBV RNA and HBcrAg contents were significantly lower than those without treatment, and the anti-HBc content was significantly higher (P<0.05). Among 18 patients with liver cirrhosis (LC), 9 were treated with NAs, the virological parameters, excepting HBV DNA, showed no significant differences between NAs-treated patients and treatment-naïve patients (P>0.05). Among 9 patients with hepatocellular carcinoma,5 were positive for serum HBV RNA (55.6%), 7 were positive for serum HBcrAg (77.8%), 8 were positive for HBV tDNA (88.9%),and 6 were positive for cccDNA (66.7%) in liver tissues. Conclusions Serum HBcrAg might be used to differentiate whether AHB is cured or not. In NAs-treated patients, even if HBV DNA decreases to the undetectable level, HBVRNA is still detectable and might cause the slow progresses of the disease, so antiviral therapy cannot be discontinued. Serum HBV RNA and HBcrAg might serve as serological substitutes for intrahepatic HBV cccDNA.