OBJECTIVE To investigate the ameliorative effects and potential mechanism of total secondary ginsenosides （TSG） on hypertrophic changes of primary cardiomyocytes stimulated by angiotensin Ⅱ （Ang Ⅱ）. METHODS Primary cardiomyocytes were isolated from the hearts of neonatal SD rats and divided into the following groups： control group， AngⅡ group （2 µmol/L）， TSG group （7.5 µg/mL）， PFK-015 group [6-phosphofructo-2-kinase/fructose-2， 6-bisphosphatase 3 （PFKFB3） inhibitor， 10 nmol/L]， and TSG+PFK-015 group （TSG 7.5 µg/mL+PFK-015 10 nmol/L）. The surface area， protein synthesis， energy metabolism-related indicators [free fatty acid （FFA）， coenzyme A （CoA）， acetyl coenzyme A （acetyl-CoA）]， and the expressions of glycolysis-related factors [hypoxia-inducible factor 1α （HIF-1α）， glucose transporter protein 4 （GLUT-4）， lactate dehydrogenase A （LDHA）， pyruvate dehydrogenase kinase 1 （PDK1） and PFKFB3] in primary cardiomyocytes of each group were measured. RESULTS Compared with the control group， the surface area of primary cardiomyocytes and protein synthesis were significantly increased， the content of FFA， protein and mRNA expressions of HIF-1α， LDHA， PDK1 and PFKFB3 were significantly increased or up-regulated in the AngⅡ group， while the contents of CoA and acetyl-CoA， the protein and mRNA expressions of GLUT-4 were significantly decreased or down-regulated （P＜0.05）. Compared with the AngⅡ group， both TSG group and PFK-015 group showed significant improvements in these indexes， with the TSG+PFK-015 group generally demonstrating superior effects compared to either treatment alone （P＜0.05）. CONCLUSIONS TSG can reduce the surface area of AngⅡ-induced primary cardiomyocytes， decrease protein synthesis， and inhibit their hypertrophic changes. These effects may be related to improving energy metabolism and the inhibition of glycolysis activity.

Combined with elastic network model (ENM), the perturbation response scanning (PRS) has emerged as a robust technique for pinpointing allosteric interactions within proteins. Here, we proposed the PRS analysis of drug-target networks (DTNs), which could provide a promising avenue in network medicine. We demonstrated the utility of the method by introducing a deep learning and network perturbation-based framework, for drug repurposing of multiple sclerosis (MS). First, the MS comorbidity network was constructed by performing a random walk with restart algorithm based on shared genes between MS and other diseases as seed nodes. Then, based on topological analysis and functional annotation, the neurotransmission module was identified as the "therapeutic module" of MS. Further, perturbation scores of drugs on the module were calculated by constructing the DTN and introducing the PRS analysis, giving a list of repurposable drugs for MS. Mechanism of action analysis both at pathway and structural levels screened dihydroergocristine as a candidate drug of MS by targeting a serotonin receptor of serotonin 2B receptor (HTR2B). Finally, we established a cuprizone-induced chronic mouse model to evaluate the alteration of HTR2B in mouse brain regions and observed that HTR2B was significantly reduced in the cuprizone-induced mouse cortex. These findings proved that the network perturbation modeling is a promising avenue for drug repurposing of MS. As a useful systematic method, our approach can also be used to discover the new molecular mechanism and provide effective candidate drugs for other complex diseases.

Objective: To analyze clinical features and electroneuromyography (ENMG) results of chronic mild occupational carbon disulfide poisoning cases. Methods: A total of 344 patients diagnosed with chronic mild occupational carbon disulfide poisoning based on GBZ 4-2002 Diagnostic Criteria of Occupational Chronic Carbon Disulfide Poisoning were selected as study subjects from 2006 to 2019 using the retrospective study method. Their clinical data was collected and analyzed. Results: The main symptoms of the study subjects were dizziness, headache, insomnia, dreaming, memory impairment, numbness and weakness in the distal extremities. Positive signs mainly included symmetrical glove and stocking distribution like sensory disorders in the distal extremities, and the weakening or absent Achilles tendon reflex and knee reflex. The incidence of symptoms and signs increased with the length of service (all P<0.01). The incidence of fundus and venous changes in patients was 41.3%, which increased with the length of service (P<0.01). ENMG examination showed varying degrees of abnormalities in the peripheral motor and/or sensory nerves in all patients, with a higher incidence of motor nerve abnormalities than sensory nerve abnormalities (21.1% vs 3.7%, P<0.01). The incidence of motor nerve abnormality was higher on the right side than the left side (23.7% vs 18.5%, P<0.01). The incidences of motor nerve abnormalities from high to low in the order were median nerve, common peroneal nerve, ulnar nerve and posterior tibial nerve (34.9% vs 27.9% vs 16.6% vs 5.1%, P<0.01). The incidences of sensory nerve abnormalities from high to low in the order were median nerve, ulnar nerve and sural nerve (5.2% vs 5.1% vs 0.7%, P<0.01). The incidences of left ulnar nerve, right ulnar nerve and right median nerve were higher in male patients than in female patients (15.2% vs 5.3%, 24.0% vs 11.7%, 44.8% vs 28.7%, all P<0.05), while the incidences of the left and right common peroneal nerve in lower extremity motor nerve were lower in male patients than in female patients (18.4% vs 52.1%, 21.2% vs 46.8%, all P<0.01). Conclusion: Chronic mild occupational carbon disulfide poisoning was mainly manifested as multiple peripheral nerve injury. ENMG results showed that the distal motor nerve conduction abnormalities were more sensitive than the sensory nerve conduction abnormalities, with a higher degree of impairment in the upper limb than the lower limb, and more impairment in the right side than the left side.

OBJECTIVE To explore the mechanism of Tingli dazao xiefei decoction on ventricular remodeling in model rats with heart failure after myocardial infarction. METHODS The rat model of heart failure after myocardial infarction was established by ligation of anterior descending branch of left coronary artery， which was divided into 8 groups： sham operation group， model group， A779 group （1 mg/kg）， A779 （1 mg/kg）+Tingli dazao xiefei decoction equivalent-dose group （0.8 g/kg）， A779 （1 mg/kg） +Tingli dazao xiefei decoction high-dose group （1.6 g/kg）， Tingli dazao xiefei decoction equivalent-dose group （0.8 g/kg）， Tingli dazao xiefei decoction high-dose group （1.6 g/kg） and losartan potassium group （10 mg/kg）. Each group was given equal volume of distilled water or corresponding drugs intragastrically for 4 weeks. Masson staining was used to determine the distribution of collagen fibers in rat myocardium. The content of hydroxyproline （Hyp） in myocardium was determined by alkaline hydrolyzation. The expressions of type Ⅰ and Ⅲ collagen （COLⅠ， COLⅢ）in myocardium were detected by immunohisto-chemistry. Myocardial fibrosis-related indexes such as matrix metalloproteinase-2 （MMP-2）， MMP-9， tissue inhibitor of metalloproteinase-1 （TIMP-1） and soluble suppression of tumorigenicity-2 （sST-2） were detected by ELISA. The protein expressions of angiotensin converting enzyme 2-angiotensin-（1-7）-Mas [ACE2-Ang-（1-7）-Mas] axis were detected by Western blot. RESULTS Compared with sham operation group， myocardial cells in model group and A779 group were disordered， collagen fiber deposition was significantly increased and myocardial fibrosis was obvious； the Hyp content and MMP-2， MMP-9， sST-2 levels were increased， and COL Ⅰ and COL Ⅲ positive expressions were significantly enhanced； TIMP-1 level， protein expressions of ACE2， Ang-（1-7） and Mas were significantly decreased （P＜0.05）. Compared with model group， above indexes of Tingli dazao xiefei decoction equivalent-dose and high-dose groups were improved to different extents. Compared with A779 group， A779+Tingli dazao xiefei decoction equivalent-dose and A779+high-dose groups could improve myocardial arrangement and collagen distribution， reduce the Hyp content and MMP-2， MMP-9 levels， reduce positive expressions of COL Ⅰ and COL Ⅲ （P＜0.05）， but couldn’t improve Ang-（1-7） and Mas protein expression. CONCLUSIONS Tingli dazao xiefei decoction can improve ventricular remodeling in myocardial failure model rats after myocardial infarction by improving the expression of ACE2- Ang（- 1-7）-Mas axis proteins.

Prevotella copri and its related taxa are widely detected in mammalian gut microbiomes and have been linked with an enterotype in humans.However,their microevolution and macroevolution among hosts are poorly characterized.In this study,extensively collected marker genes and genomes were analyzed to trace their evolutionary history,host specificity,and biogeographic distribution.Investigations based on marker genes and genomes suggest that a P.copri-containing lineage(PCL)harbors diverse species in higher primates.Firstly,P.copri in the human gut consisted of multiple groups exhibiting high genomic divergence and conspicuous but non-strict biogeographic patterns.Most African strains with high genomic divergence from other strains were phylogenetically located at the root of the species,indicating the co-evolutionary his-tory of P.copri and Homo sapiens.Secondly,although long-term co-evolution between PCL and higher primates was revealed,sporadic signals of co-speciation and extensive host jumping of PCL members were suggested among higher primates.Metagenomic and phylogenetic analyses indicated that P.copri and other PCL species found in domesticated mammals had been recently transmitted from humans.Thirdly,strong evidence was found on the extensively horizontal transfer of genes(e.g.,genes encoding carbohydrate-active enzymes)among sympatric P.copri groups and PCL species in the same primate host.Our study provides panoramic insights into the combined effects of vertical and horizontal transmission,as well as potential niche adaptation,on the microevolutionary and macroevolutionary history for an enterotype-representative lineage.

Objective:To understand the epidemiological characteristics of rabies in China from 2007 to 2018, and to provide reference information for formulating strategies to eliminate rabies.Methods:The incidence data were obtained from the national notifiable communicable disease reporting information system in China to analyze the spatial, population and time distribution of human rabies cases.Results:From 2007 to 2018, 17 848 cases of human rabies were reported nationwide. 57% of the cases were reported in 6 provinces, including Guangxi, Guangdong, Guizhou, Hunan, Henan and Sichuan. A total of 37 counties reported more than 50 cases. The number of reported cases decreased from 3 300 in 2007 to 422 in 2018, with an average annual decline rate of 7%. The number of districts and counties involved decreased from 984 in 2007 to 308 in 2018. The population aged 35-75 years accounted for 67%, followed by the population aged 0-15 years (17%). Farmers accounted for 70% of the total number of cases, followed by students (11%) and scattered children (6%).Conclusions:China′s human rabies epidemic has changed from a regional epidemic to a multi-regional spread.

Objective    To explore the effects and molecular mechanisms of histone methylase G9a inhibitor BIX-01294 on apoptosis in esophageal squamous cell carcinoma (ESCC). Methods    MTT assay and Colony-forming Units were adopted to determine the effects of BIX-01294 on the growth and proliferation of ESCC cell lines EC109 and KYSE150. Flow cytometry was used to analyze the apoptosis status of ESCC cells after the treatment of BIX-01294. The effects of BIX-01294 treatment on the expressions of G9a catalytic product H3K9me2, DNA double-strand break (DSB) markers, and apoptosis-related proteins were detected by Western blotting. Results    BIX-01294 inhibited the growth of EC109 and KYSE150 cells in a dose-dependent manner (P<0.05), and BIX-01294 with the inhibitory concentration 50%(IC50) significantly inhibited the formation of colony (P<0.05). After 24 hours treatment of BIX-01294 (IC50), the apoptosis rate of EC109 cells increased from 11.5%±2.1% to 42.5%±5.4%, and KYSE150 cells from 7.5%±0.9% to 49.2%±5.2%(P<0.05). The expression level of the G9a catalytic product, H3K9me2, significantly decreased (P<0.05); while the expression of the DSB marker γH2AX was dramatically enhanced (P<0.05). We also found that the mitochondrial apoptosis pathway was activated and the expression levels of cleaved caspase3 and cleaved PARP were significantly elevated (P<0.05). Conclusion    BIX-01294, the inhibitor of methyltransferase G9a, prompted apoptosis in ESCC cells by inducing DSB damage and activating mitochondrial apoptosis pathway.

Objective:To investigate the short-term therapeutic effect and long-term survival of multiple myeloma patients with extramedullary disease (EMD) in the new drug era.Methods:The data of 74 patients with multiple myeloma diagnosed and treated in Anhui Wanbei Coal and Electricity Group General Hospital from January 2015 to January 2020 were retrospectively analyzed, including 17 patients with soft tissue infiltration (EM-S), 9 patients with bone infiltration (EM-B), and 48 patients without EMD (No-EMD). The short-term efficacy, the 4-year progression-free survival (PFS) rate and overall survival (OS) rate, and their influencing factors in three groups of patients after receiving bortezomib regimen were analyzed.Results:After 3-4 courses of early induction therapy of bortezomib regimen, the overall response rate of patients in the EM-S group was lower than that in the No-EMD group and the EM-B group [58.8% (10/17) vs. 85.4% (41/48), 100.0% (9/9)], and the differences were statistically significant ( χ2 = 13.7, P = 0.036; χ2 = 26.5, P = 0.003), while the difference between No-EMD group and EM-B group was not statistically significant ( χ2 = 12.7, P = 0.211). Survival analysis showed that the 4-year PFS rate of No-EMD group was higher than that of the EM-S group and EM-B group (41.0% vs. 7.6%, 0), and the differences were statistically significant ( χ2 = 10.835, P < 0.01; χ2 = 8.276, P = 0.004). Meanwhile, the 4-year OS rate of EM-S group was lower than that of the No-EMD group and EM-B group (16.5% vs. 54.3%, 59.3%), and the differences were statistically significant ( χ2 = 9.146, P = 0.002; χ2 = 4.066, P = 0.044). Conclusion:The early treatment effect of bortezomib regimen, PFS and OS in multiple myeloma patients with EM-S are poor, while the EM-B has no effect on OS.

Background/Aims: To investigate postpartum hepatic flares and associated factors in highly viremic pregnant patients in the immune tolerance phase who adopted telbivudine (LdT) treatment in the last trimester to reduce vertical transmission of hepatitis B virus. Methods: Hepatitis B e antigen (HBeAg)-positive, highly viremic pregnant women were recruited for this prospective study. Treatment with LdT was started from 28 weeks of gestation. Virological and biochemical markers were examined before LdT treatment, antepartum and postpartum. Serial blood samples at the same time were collected to detect cytokines and cortisol (COR). Results: Fifty-six of 153 patients (36.6%) had postpartum hepatic flares, defined as a 2-fold increase in alanine aminotransferase 6 weeks after delivery. Age and the antepartum alanine aminotransferase and postpartum HBeAg levels were independent influencing factors of postpartum hepatic flares. Cytokines showed no regularity during or after pregnancy. Compared with the patients with no postpartum flares, the patients with flares had lower baseline interferon γ and COR levels (p=0.022 and p=0.028) and higher postpartum interferon γ levels (p=0.026). Conclusions A high proportion of highly viremic and immune-tolerant pregnant patients treated with LdT in the last trimester had postpartum hepatic flares, which implied that these patients entered the immune clearance phase after delivery. Thus, this may create an appropriate opportunity for re-antiviral therapy.

Objective:To investigate the efficacy and safety of transcatheter arterial chemoembolization (TACE) combined with sorafenib and sequential microwave ablation (MWA) versus TACE combined with sorafenib in the treatment of hepatocellular carcinoma (HCC) with tumor diameter over 5 cm, and to analyze risk factors affecting the prognosis of patients.Methods:The prospective cohort study was conducted. The clinicopathological data of 61 HCC patients with tumor diameter over 5 cm who were admitted to two medical centers (30 in the Laiyang Central Hospital of Yantai City Affiliated to Weifang Medical College and 31 in the Qingdao Central Hospital Affiliated to Qingdao University) between July 2012 and November 2013 were collected. Patients who were treated with TACE combined with sorafenib and sequential MWA were allocated into observation group, and patients who were treated with TACE combined with sorafenib were allocated into control group. Observation indicators: (1) treatment, complications and adverse drug reactions; (2) short-term efficacies; (3) follow-up and survival situations; (4) analysis of prognostic factors. Follow-up was performed by inpatient, outpatient examinations or telephone interview once a month within the first 6 months after treatment and once every 3 months thereafter up to November 2018. The follow-up included laboratory indicators, tumor markers, abdominal enhanced computed tomography or magnetic resonance imaging examinations. The survival of patients and disease progression were fully documented. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was performed by the t test. Measurement data with skewed distribution were described as M (range), and comparison between groups was performed using the Wilcoxon rank sum test. Count data were expressed as absolute numbers or percentages, and comparison between groups was performed using the chi-square test or pearson-corrected chi-square test. Ranked data were analyzed using the Wilcoxon rank sum test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves. The Log-rank test was used for survival analysis. Comparison of survival rates between time points was performed using the Bonferroni method to adjust the test level. Univariate and multivariate analyses were performed using the multiple COX proportional hazard model. Results:A total of 61 HCC patients were selected for eligibility, including 36 males and 25 females, aged (58±8)years, with a range from 43 to 73 years. Of the 61 patients, 31 were in the observation group and 30 in the control group. (1) Treatment, complications and adverse drug reactions: ① treatment information. The treatment times of TACE, treatment times of MWA, time from the first TACE to the first sorafenib medication, and duration of sorafenib medication in the observation group were 1 time (range, 1-5 times), 2 times (range, 1-4 times), 5 days (range, 5-9 days), 24 months (range, 6-72 months), respectively. The above indicators of patients in the control group were 3 times (range, 1-5 times), 0, 6 days (range, 5-9 days), and 16 months (range, 6-60 months). There were significant differences in the treatment times of TACE, treatment times of MWA, and duration of sorafenib medication between the two groups ( Z=4.701, -7.213, -2.614, P<0.05). There was no significant difference in the time from the first TACE to the first sorafenib medication between the two groups ( Z=0.573, P>0.05). ② Complications: there was no TACE related complications in the two groups. There were 3 patients with MWA related complications in the observation group, including 2 cases of minor hemorrhage under the liver capsule and 1 case of pleural effusion, and they were relieved after conservative treatment. ③ Adverse reactions to sorafenib: after 2 months of sorafenib medication, patients in the observation group and control group had at least one kind of sorafenib related stage Ⅰ-Ⅲ adverse reaction, without stage Ⅳ adverse reaction. The numbers of cases with hand-foot skin reaction, rash, pruritus, loss of skin pigmentation, diarrhea, decreased appetite, nausea and vomiting, pain in the liver area, fever, fatigue, liver dysfunction, bone marrow suppression were 8, 3, 4, 3, 10, 18, 20, 20, 20, 15, 3, 2 in the observation group, and 9, 3, 3, 2, 13, 19, 23, 12, 21, 12, 6, 2 in the control group, respectively, showing no significant difference in the above indices between the two groups ( χ2=0.133, 0.000, 0.000, 0.000, 0.796, 0.177, 1.082, 3.674, 0.208, 0.435, 0.601, 0.000, P>0.05). Patients with adverse reactions to sorafenib were relieved by symptomatic treatment, reducing the dose of sorafenib or intermittent drug withdrawal. (2) Short-term efficacies: the level of alpha fetoprotein was 16 μg/L (range, 3-538 μg/L) in the observation group and 292 μg/L (range, 9-642 μg/L) in the control group after one month of treatment, showing a significant difference between the two groups ( Z=3.744, P<0.05). After 3 months of treatment, cases with tumor complete remission, cases with tumor partial remission, cases with stable disease, cases with progressive disease, objective response rate, and disease control rate in the observation group were 14, 11, 6, 0, 80.6%(25/31) , 100.0%(31/31), respectively. The above indicators in the control group were 2, 13, 12, 3, 50.0%(15/30), 90.0%(27/30). There was a significant difference in the objective response rate between the two groups ( χ2=6.343, P<0.05), but no significant difference in the disease control rate between the two groups ( χ2= 1.473, P>0.05). (3) Follow-up and survival situations: 61 HCC patients were followed up for 9.0-75.0 months, with a median follow-up time of 22.0 months. During the follow-up, 28 patients in the observation group had progressive disease, including 8 cases of local tumor progression, 4 of portal vein tumor thrombi, 11 of intrahepatic metastasis, and 5 of pulmonary metastasis. Thirty patients in the control group had progressive disease, including 13 cases of local tumor progression, 6 of portal vein tumor thrombi, 6 of intrahepatic metastasis, and 5 of pulmonary metastasis. Among the 61 patients, 28 patients in the observation group and 29 patients in the control group died. The median overall survival time and median progression-free survival time of the observation group was 28.0 months and 18.0 months, versus 19.5 months and 11.5 months of the control group, showing significant differences between the two groups ( χ2=8.021, 10.506, P<0.05). The 1-, 3-, 5-year overall survival rates of the observation group were 97%, 37% and 20%, respectively, versus 83%, 13% and 7% of the control group, showing significant differences in the above indicators between the two groups ( Z=23.635, 4.623, 3.139, P<0.0167). The 1-, 2-, 3-year progression-free survival rates of the observation group were 77%, 40%, and 27%, respectively, versus 43%, 13%, and 7% of the control group, showing significant differences in the above indicators between the two groups ( Z=9.965, 4.900, 3.684, P<0.0167). (4) Analysis of prognostic factors: results of univariate analysis showed that treatment method, maximum tumor diameter, Barcelona clinical liver cancer (BCLC) stage, liver cirrhosis, hepatitis B virus(HBV) infection, and Child-Pugh classification were related factors for overall survival time [ hazard ratio ( HR)=0.483, 6.196, 12.646, 5.049, 2.950, 4.791, 95% confidence interval ( CI): 0.284-0.823, 3.198-12.003, 5.031-31.785, 2.586-9.858, 1.366-6.369, 2.507-9.155, P<0.05] and progression-free survival time ( HR=0.427, 5.804, 7.032, 5.405, 2.925, 4.410, 95% CI: 0.248-0.735, 3.043-11.070, 3.071-16.101, 2.685-10.881, 1.364-6.270, 2.331-8.342, P<0.05). Results of multivariate analysis showed that treatment method, maximum tumor diameter, BCLC stage, liver cirrhosis, and HBV infection were independent influencing factors for overall survival time ( HR=0.183, 5.886, 17.544, 4.702, 3.801, 95% CI: 0.090-0.370, 2.648-13.083, 5.740-53.622, 1.928-11.470, 1.368-10.562, P<0.05) and progression-free survival time ( HR=0.201, 3.850, 3.843, 3.598, 3.726, 95% CI: 0.098-0.411, 1.761-8.414, 1.526-9.682, 1.444-8.963, 1.396-9.947, P<0.05). Conclusions:Compared with TACE combined with sorafenib, TACE combined with sorafenib and sequential MWA is safe and effective in the treatment of HCC with tumor diameter over 5 cm. The treatment method, maximum tumor diameter, BCLC stage, liver cirrhosis, and HBV infection are independent influencing factors for overall survival time and progression-free survival time of patients.