In order to study whether marrow stromal cells (MSCs) can be induced into nerve-like cells in vitro, and the mechanism, the MSCs in Wistar rats were isolated and cultured, and then induced with DMSO and BHA in vitro. The expression of specific marking proteins in neurons, glia and neural stem cells were detected before preinduction, at 24 h of preinduction, at 6 h, 24 h, and 48 h of neuronal induction by using immunohistochemistry and Western blotting. The ultrastructural changes after the inducement were observed. The results showed that after the inducement, many MSCs turned into bipolar, multipolar and taper, and then intersected as network structure. At the same time, some MSCs had the typical neuron-like ultrastructure. Immunohistochemistry revealed that NeuN and Nestin expression was detectable after inducement, but there was no GFAP and CNP expression. Western blotting showed the expression of Nestin was strong at 6 h of neuronal induction, and decreased at 24 h, 48 h of the induction. NeuN was detectable at 6 h of neuronal induction, and increased at 24 h, 48 h of the induction. It was concluded MSCs were induced into neural stem cells, and then differentiated into neuron-like cells in vitro.
Bone Marrow Cells/*cytology ; *Cell Differentiation ; Cells, Cultured ; Glial Fibrillary Acidic Protein/metabolism ; Neurons/*cytology ; Rats, Wistar ; Stromal Cells/cytology

ObjectiveTo study the mechanism of differentiation of mesenchymal stem cells(MSCs) into neuron-like cells in vitro.MethodsMSCs of Wistar rats were separated and cultured, and then induced with DMSO and BHA in vitro. The specific marking proteins of neurons, glia and neural stem cells were detected before preinduction, at 24h after preinduction, at 6h, 24h, and 48h after neuronal induction.ResultsAfter the inducement, many MSCs turned into bipolar,multipolar and taper,and then intersected as network structure. Nestin was strong positive at 6h after neuronal induction, and decreased at 24h, 48h after the induction. NeuN was present at 6 h after neuronal induction, and increased at 24h, 48h after the induction.ConclusionMSCs can be induced into neural stem cells(NSCs) at first, and then differentiate into neuron-like cells in vitro.

Purpose:Thalamic hemorrhage breaking into ventricles (THBIV) is a devastating disease with high morbidity and mortality rates.Endoscopic surgery (ES) may improve outcomes,although there is no consensus on its superiority.We investigated the efficacy and safety of ES and compared the outcomes of different management strategies by ES,hematoma puncture and drainage (HPD),and external ventricular drainage (EVD) in patients with THBIV.Methods:We retrospectively analyzed patients with THBIV treated by ES,HPD,or EVD at our hospital from June 2015 to June 2018.Patients were categorized into anteromedial and posterolateral groups based on THBIV location,and then the two groups were further divided into ES,HPD,and EVD subgroups.Individualized surgical approach was adopted according to the location of the hematoma in the ES subgroups.Patient characteristics and surgical outcomes were investigated.Results:We analyzed 211 consecutive patients.There were no significant differences in clinical characteristics or incidence of perioperative procedure-related complications (postoperative rebleeding and intracranial infection) in either anteromedial or posterolateral groups.Compared with other therapeutic methods,the ES subgroups had the highest hematoma evacuation rate,shortest drainage time,and lowest incidence of chronic ventricular dilatation (all p ＜ 0.05).Among the three anteromedial subgroups,ES subgroup had the best clinical outcomes which was assessed by the modified Rankin Scale,followed by HPD and EVD subgroups (p ＜ 0.01);while in the posterolateral subgroups,clinical outcomes in the ES and HPD subgroups were similar and better than that in the EVD subgroup (p =0.037).Conclusion:Individualized surgical ES approach for removal of thalamic and ventricular hematomas is a minimally invasive,safe,and effective strategy for the treatment of THBIV with a thalamic hematoma volume of 10-30 mL.

Objective A novel multiplex real-time RT-PCR kit was developed to detect EV71,CoxA16 and other human enteroviruses simultaneously with an internal amplification control to avoids false negatives,which used for hand,foot and mouth disease in the clinical diagnosis and epidemiological surveillance.Methods Design specific primers and probes of EV71,CA16,other intestinal virus and internal amplification control,improve the extraction method of virus nucleic acid.Optimization the detection system of real-time quantitative PCR.Research the products of the accuracy,stability,precision,amplification efficiency and detection of linear range.Results The primers and probes had high spicificity.The Viral RNA extraction effect of this Kit is as same as that of QIAamp Viral RNA mini Kit (QIAGEN company),but less reagent cost.The optimal concentrations of primers and probes are 0.2 μmol/L for all the upstream and downstream primers,0.06 μmol/L for probes of other human enteroviruse,0.08 μmol/Lfor probes of EV71 and CA16 respectively.The kit has good stability,accuracy and precision.The amplification efficiencies of EV71,CoxA16 and other human enteroviruses are 106％,101％ and 105％ and the detection of linear range is from 109 copies/μl-102 copies/μl.Conclusion The novel multiplex realtime RT-PCR kit for detecting EV71,CoxA16 and other human enteroviruses simultaneously with an internal amplification control has good stability,accuracy,precision and amplification efficiencies.So it has great value in clinical application.

In order to study whether marrow stromal cells (MSCs) can be induced into nerve-like cells in vitro, and the mechanism, the MSCs in Wistar rats were isolated and cultured, and then induced with DMSO and BHA in vitro. The expression of specific marking proteins in neurons, glia and neural stem cells were detected before preinduction, at 24 h of preinduction, at 6 h, 24 h, and 48 h of neuronal induction by using immunohistochemistry and Western blotting. The ultrastructural changes after the inducement were observed. The results showed that after the inducement, many MSCs turned into bipolar, multipolar and taper, and then intersected as network structure. At the same time, some MSCs had the typical neuron-like ultrastructure. Immunohistochemistry revealed that NeuN and Nestin expression was detectable after inducement, but there was no GFAP and CNP expression. Western blotting showed the expression of Nestin was strong at 6 h of neuronal induction, and decreased at 24 h, 48 h of the induction. NeuN was detectable at 6 h of neuronal induction, and increased at 24 h, 48 h of the induction. It was concluded MSCs were induced into neural stem cells, and then differentiated into neuron-like cells in vitro.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Glial Fibrillary Acidic Protein ; metabolism ; Neurons ; cytology ; Rats ; Rats, Wistar ; Stromal Cells ; cytology

OBJECTIVETo investigate the expression characteristics of nuclear factor kappa B (NF-kB) in hepatocellular carcinoma (HCC) tissues and its correlation with tumor necrosis factor alpha (TNF alpha) and clinical pathological features.

METHODSThirty liver specimens from HCC patients were collected by self-control method. The localization and expression of NF-kappaB in HCC and their surrounding tissues were detected by immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively. And the levels of TNF alpha in these tissues were analyzed by ELISA.

RESULTSThe expressed NF-kappaB was localized in nucleus and cytoplasm in HCC, whereas only in cytoplasm in the surrounding tissues. The expression level and density of NF-kappaB in HCC tissues were obviously higher than those in the surrounding tissues (P < 0.01), which was positively correlated with increased TNF alpha in HCC tissues (r = 0.964, P < 0.01). No positive correlation was found between NF-kappaB expression and histological differentiation grade, number of tumor, size of tumor, and HBsAg positive (P > 0.05).

CONCLUSIONThe expression and localization of NF-kappaB in HCC tissues are obviously different from those in the surrounding normal liver tissues, and the level of nucleoprotein NF-kappaB in HCC tissues is correlated with expressed TNF alpha, suggesting that TNF alpha can activate NF-kB, the activated NF-kB then translocates to the nucleus and plays important role in the carcinogenesis of HCC.

Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Nucleus ; metabolism ; Female ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism

Objective To study the variants of S gene of hepatitis B virus among the breakthrough infection children immunized with hepatitis B vaccine in Huzhou city.Methods HBV DNA S gene regions were amplified and sequenced in 29 children with HBsAg-positive/HBV DNA-positive after immunized with hepatitis B vaccine.Results The sequencing results showed that 26 cases were B genotypes,3 were C genotypes.The amino acids mutations were presented in 9 children serum HBV DNA.The major amino acid mutations were in the "a" determinant of HBV DNA S gene.Conclusion Different S gene variants maybe cause HBV vaccination failure in some children in Huzhou city.No predominant variant was found.

OBJECTIVETo study the correlations between preS1 antigen, HBV-DNA and hepatitis B virus (HBV) serum markers in patients with chronic hepatitis B.

METHODSThe HBV markers, preS1 antigen and HBV-DNA were determined using enzyme- linked immunosorbent assay and quantitative PCR in 1158 patients with chronic hepatitis B.

RESULTSIn these patients, the HBV-DNA positivity rate was 68.9%, significantly higher than preS1 antigen positivity (54.8%, chi2=53.24, P<0.005). The positivity rates of both HBV-DNA and PreS1-antigen were significantly higher in HBeAg-positive patients than in HBeAg-negative patients (P<0.005). The coincident rates of preS1-antigen and HBeAg with HBV-DNA were 56.9% and 63.3%, respectively. PreS1 antigen had higher sensitivity but lower specificity than HBeAg. The detection rates of preS1 antigen and HBeAg increased with the level of HBV-DNA, and preS1 antigen positivity was higher than that of HBeAg in patients with low HBV-DNA levels.

CONCLUSIONDetection of HBV serum markers along with preS1 antigen and HBV-DNA may help assess the status of viral replication and therapeutic efficacy in patients with chronic hepatitis B. PreS1 antigen may serve as an auxiliary indicator in HBeAg-negative cases or when HBV-DNA detection is impossible.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; DNA, Viral ; blood ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; isolation & purification ; Hepatitis B, Chronic ; blood ; immunology ; virology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; Virus Replication ; genetics ; Young Adult

Purpose:: Thalamic hemorrhage breaking into ventricles (THBIV) is a devastating disease with high morbidity and mortality rates. Endoscopic surgery (ES) may improve outcomes, although there is no consensus on its superiority. We investigated the efficacy and safety of ES and compared the outcomes of different management strategies by ES, hematoma puncture and drainage (HPD), and external ventricular drainage (EVD) in patients with THBIV. Methods:: We retrospectively analyzed patients with THBIV treated by ES, HPD, or EVD at our hospital from June 2015 to June 2018. Patients were categorized into anteromedial and posterolateral groups based on THBIV location, and then the two groups were further divided into ES, HPD, and EVD subgroups. Individualized surgical approach was adopted according to the location of the hematoma in the ES subgroups. Patient characteristics and surgical outcomes were investigated. Results:: We analyzed 211 consecutive patients. There were no significant differences in clinical characteristics or incidence of perioperative procedure-related complications (postoperative rebleeding and intracranial infection) in either anteromedial or posterolateral groups. Compared with other therapeutic methods, the ES subgroups had the highest hematoma evacuation rate, shortest drainage time, and lowest incidence of chronic ventricular dilatation (all p < 0.05). Among the three anteromedial subgroups, ES subgroup had the best clinical outcomes which was assessed by the modified Rankin Scale, followed by HPD and EVD subgroups (p < 0.01); while in the posterolateral subgroups, clinical outcomes in the ES and HPD subgroups were similar and better than that in the EVD subgroup (p = 0.037). Conclusion: Individualized surgical ES approach for removal of thalamic and ventricular hematomas is a minimally invasive, safe, and effective strategy for the treatment of THBIV with a thalamic hematoma volume of 10-30 mL.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.