Gastric cancer is the third leading cause of death from malignancy and the fifth most common malignancy in the world. The development and progression of intestinal-type gastric cancer is in accordance with the "Correa model", in which gastric mucosal intestinal metaplasia (GIM) is the key link in the transformation of the gastric mucosa from benign to malignant and is also one of the most common precancerous lesions of gastric cancer, therefore, it is the key to investigate the pathogenesis of GIM and intervene it at an early stage for the prevention and treatment of gastric cancer. Helicobacter pylori (HP) infection is considered as one of the recognized risk factors for GIM. However, GIM cannot be reversed by eradication of HP alone, considering that there may be other factors that continue to play a role in the development and progression of GIM. In recent years, it has been confirmed in several studies that bile reflux can induce GIM, but the specific molecular mechanism by which bile reflux induces GIM is not clear.The mechanism of bile reflux induced gastric mucosal injury, the relationship between GIM and gastric cancer, the research status of bile reflux induced GIM, and the molecular mechanisms of bile acid induced GIM are discussed in this article, aiming to improve clinicians' understanding of GIM induced by bile reflux and provide a basis for the early intervention of GIM and prevention of gastric cancer.

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.

Chronic hepatitis B virus infection is prevalent worldwide, and can progress to liver cirrhosis and even hepatocellular carcinoma if left untreated. Over the past 20 years, due to the approval and upgrading of nucleoside/nucleotide analogues, a qualitative leap has been made in inhibiting HBV replication. Most chronic hepatitis B (CHB) patients can achieve relief of hepatocellular inflammation, regression of hepatic fibrosis, improvement of life quality and survival time. With the extension of treatment time and the enrichment of treatment experience, new hot clinical issues gradually emerge, including the clinical application of new serum markers, the expansion of indicators for CHB antiviral treatment, the formation of CHB treatment plan and the improvement of long-term prognosis, which derserve further clinical attention.

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 identify the high-risk factors and establish a nomogram for evaluating significant histological response (SHR) in chronic hepatitis B (CHB) patients receiving entecavir treatment. Methods Treatment-naive CHB patients who were presented to 14 hospitals, from October 2013 to October 2014, were enrolled and treated with entecavir for 72 weeks,prospectively. All the patients who underwent paired biopsies at treatment baseline and week 72 were analyzed. According to whether SHR (Ishak fibrosis score F≤2 points and histology activity index HAI≤4 points) was obtained during treatment, they were assigned to response group (n=160) and non-response group (n=567). High-risk factors were identified by multivariate logistic regression, and then were incorporated into a nomogram model. The discrimination, calibration and clinical applicability of nomogram were assessed by concordance index (C-index), calibration curve and clinical decision curve (DCA). Results After 72 weeks of treatment, regression of fibrosis, improvement of inflammation, virologic response, alanine aminotransferase (ALT)normalization and HBeAg seroconversion were 51.2%, 74.4%, 86.0%, 83.5% and 13.3%, respectively, however, 49.0% (306/625) of patients with virological response and 43.4% (165/380) of patients with ALT normalization did not achieved regression of fibrosis.Logistic regression analysis showed that baseline age (OR=0.978, 95%CI 0.958-0.998, P=0.030), platelet (PLT) (OR=1.005, 95%CI 1.001-1.010, P=0.030), liver stiffness measurement (LSM) (OR=0.931, 95%CI 0.892-0.972, P=0.001) and 72-week ALT (OR=0.980,95%CI 0.964-0.996, P=0.016), 72-week LSM (OR=0.858, 95%CI 0.782-0.941, P=0.001) were independent high-risk factors associated with SHR. The C-index of the nomogram model based on the above factors was 0.784, which was significantly better than 72-week AST/PLT ratio (APRI) (0.643), fibrosis-4 (FIB-4) (0.691) and LSM (0.735) alone, and had well-fitted calibration curves and DCA. Conclusions Incorporating baseline age, PLT, LSM, 72-week ALT and 72-week LSM, the established individualized nomogram model for evaluating significant histological response in CHB patients receiving antiviral therapy has good predictive performance and can reduce the need of liver biopsy.

Objective To analyze the risk factors of esophago-gastric fundal varices (EGV) in patients with hepatitis B virus (HBV)-related compensated cirrhosis, and to compare the clinical performance of multiple noninvasive prediction models for EGV evaluation. Methods A retrospective analysis was performed on the clinical data of patients with hepatitis B cirrhosis from two centers from January 2017 to December 2019. All patients underwent biochemical examination, gastroscopy, and liver stiffness measurement (LSM). AST-to-platelet ratio index (APRI), Fibrosis 4 index (FIB-4) and LSM-spleen diameter (SD)-to-platelet ratio score (LSPS) were calculated. According to the results of gastroscopy, they were divided into EGV and non-EGV group. The high-risk factors of EGV were explored by binary logistic regression. The receiver operating characteristic curve (ROC) was plotted, and the area under the curve (AUC) was calculated to evaluate the clinical performance of each indicator in predicting EGV. Results A total of 470 patients were enrolled, including 292 in the non-EGV group and 178 in the EGV group. Logistic regression analysis showed low PLT (platelet)(OR=0.99, 95%CI 0.98-0.99), high SD (spleen diameter)(OR=1.02, 95%CI 1.01-1.03) and high LSM(OR=1.04, 95%CI 1.02-1.07) were high risk factors for EGV (all P<0.05). The AUC of LSPS in predicting EGV [0.75(95%CI 0.71-0.79)] was significantly higher than that of PLT [0.72(95%CI 0.67-0.76)], SD [0.69(95%CI 0.64-0.73)], LSM [0.67(95%CI 0.63-0.72)], APRI [0.69(95%CI 0.65-0.74)] and FIB-4 [0.66(95%CI 0.62-0.71)](P<0.05). The LSPS score in EGV group was significantly higher than that in non-EGV group [2.7(1.3, 5.0) vs. 1.0(0.5, 1.7), P<0.001]. With the aggravation of EGV, LSPS score showed an upward trend (r=0.426, P<0.001). The cutoff value of LSPS for high risk of EGV was >3.5, the corresponding specificity and positive predictive value (PPV) ratio were 93.6% and 79.8% respectively. The cutoff value of LSPS for low risk of EGV was<1.3, its sensitivity and negative predictive value were 75.5% and 81.1% respectively. Conclusions Incorporating three risk factors including low PLT, high LSM and SD, the LSPS in predicting EGV in patients with compensated hepatitis B cirrhosis is better than ARPI, FIB-4, or LSM, and can reduce the need for gastroscopy.

Objective To investigate the effect of non-alcoholic fatty liver disease (NAFLD) on the overall survival of patients with hepatitis B virus-related hepatocellular carcinoma (HCC). Methods The hospitalized patients from June 2008 to December 2020 were screened according to inclusive and exclusive criteria, the clinical characteristics at the time of the initial diagnosis of HCC and time-to-event information of the enrolled patients were retrospectively collected. After stratified by the Barcelona Clinic Liver Cancer (BCLC) staging and then balanced with the stabilized inverse probability of treatment weighting(IPTW) between NAFLD and non-NAFLD group, the Kaplan-Meier survival curve, log-rank test and Cox regression were used to compare the prognosis of patients with HBV-related HCC between NAFLD group and non-NAFLD group. The endpoint of this study was HCC-related death. Results A total of 833 patients were enrolled in the study. Among them, 465 patients in resectable group and 368 in non-resectable group. There were 161 (34.6%) patients with NAFLD in resectable group, and 76 (20.7%) patients with NAFLD in non-resectable group. After stabilized IPTW weighting, all indicators showed no statistical difference between two groups. Survival analyses showed that median survival time of NAFLD patients with HBV-related HCC was significantly longer than that of non-NAFLD patients. The 3-, 5-, and 8-year cumulative survival rates of NAFLD patients in resectable group were 96.9%, 82.1% and 41.2%, which were significantly higher than 91.1%, 60.4% and 13.7% of non-NAFLD patients (P<0.01). Cox analysis showed that NAFLD was the protective factor of survive (HR=0.473, 95%CI 0.356-0.627, P<0.01). Similarly, those of NAFLD patients in non-resectable group were 76.8%, 53.7% and 18.4%, which were significantly higher than those of non-NAFLD patients (53.0%, 25.7% and 2.5%, respectively, P<0.01). Cox analysis also showed that NAFLD was the protective factor of survive(HR=0.358, 95%CI 0.247-0.518, P<0.01). Conclusion The cumulative survival rate of HBV-related HCC patients with NAFLD is higher than that of patients without NAFLD.

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.

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.

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.