The primary neural stem cells were isolated from SD rat and formed the neuropheres, the neuropheres were passaged and planted on the dish coated with 0.1% gelatin, the colony was picked up under the microscope, then dispersed and cultured, to obtain the clone proliferated from one cell, passaging and picking up the cells 5～6 times at least. The NSC and its differentiated cells were identified with the marker genes respectively. The results showed that the neural stem cells were isolated from the SD rat embryos and the real clone were obtained by picking up the cells again and again, and then cultured in the form of monolayer. The marker genes of the neural stem cells and its differentiated cells could be detected at last. It will provide the rat model the resource of the cells for the treatment and the basic research for the morphology standard.

Objective To explore the impact of broad antigen HLA-Bw4 on disease progression in HIV-1 infected subjects. Methods Three hundred and forty subjects chronically infected with HIV-1 and 69 HIV-1 negative subjects were recruited and HLA-B alleles were typed with sequence-based high resolution typing assay. HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD4+ T cell counts and viral loads were analyzed. Results Sixty-five HLA-B alleles were detected in HIV-1 positive subjects. Subjects with Bw4 (Bw4 homozygotes and Bw4Bw6 heterozygotes ) had higher CD4+ T cell counts ( P = 0. 004 ) and lower plasma viral load ( P = 0.003 ) than subjects without Bw4 ( Bw6 homozygotes). When compared with HIV-1 postive subjects with CD4+ T cell counts above 500 celis/μl, those with CD4+ T cell counts below 500 cells/μl were observed with decreased percentage of Bw4Bw6 heterozygote ( P =0.0002) and increased percentage of Bw6 homozygotes ( P < 0. 0001 ). There is no significant difference in CD4+ T cell counts between Bw4 homozygotes and Bw4Bw6 heterozygote, but lower viral loads were observed in Bw4Bw6 heterozygotes( P = 0. 037 ). Conclusion HLA-Bw4 can confer pretective effects on H1V-1 infected subjects by maintaining higher CD4+ T cell counts and lower viral load, the mechanism behind this effect need further exploration.

Objective Folate receptors ( FRs) , overexpressed on the surface of a variety of tumor cells , are potential targets for tumor targeting therapy .This study aimed to prepare an FR-mediated drug nanocarrier with folate conjugated pullulan acetate ( FPA) to target chemotherapeutic agents to FR-overexpressed tumor cells and investigate its tumor-suppressing effect in vitro. Methods The cytotoxicity of epirubicin-loaded FPA nanoparticles ( FPA/EPI NP) against HepG2 and Hela cells was evaluated by MTS assay.The HepG2 and Hela cells were divided into five groups to be treated with NPs (NP control), chlorpromazine, chloro-quine, amiloride, and folate, respectively, followed by detection of the fluorescence intensity by flow cytometry . Results FPA/EPI NP was successfully formulated into NPs , with the mean particle size of (268.5 ±12.0) nm, by dialysis with an almost spherical shape . The drug-loading rate and entrapment rate of FPA/EPI NP were (6.45 ±1.04) and (72.45 ±11.50) %, respectively.The survival rates of the HepG2 and Hela cells were both >95%after 24 hours of incubation with FPA NP at 5, 40, 200, 400 and 1000μg/mL and 90.0%after 72 hours.The survival rates of the HepG2 cells treated with 5, 40, 200, 400 and 1000μg/mL FPA/EPI NP for 24 hours were (92.3 ±5.2), (70.4 ±4.6), (50.0 ±4.0), (41.1 ±4.1) and (27.0 ±3.6) %, respectively.Compared with the NP control group, the Hela cells of the chlorpromazine , amiloride and folate groups showed a significantly lower rate of NP uptake (P<0.05), and so did the HepG2 cells pretreated with chlorpromazine or amilo-ride (P<0.05).At 72 hours, the half maximal inhibitory concentrations (IC50) of FPA/EPI NP against HepG2 and Hela cells were 168 and 105μg/mL, respectively . Conclusion Clathrin-mediated endocytosis and macropinocytosis are involved in the internaliza-tion of FPA/EPI NP in HepG2 cells, while clathrin-and FR-mediated endocytosis in that of Hela cells .FPA NP may serve as a new drug carrier for tumor-targeted therapy .

OBJECTIVE:To compare the economics of interferon α1b and α2b in the treatment of chronic hepatitis C. METH-ODS:By retrospective study,114 patients with chronic hepatitis C who received interferon were selected,60 patients received interfer-on α1b were divided into group A and 54 patients received interferon α2b were divided into group B. Negative conversion rate of HCV-RNA,normalization rate of ALT and the incidence of ADR in 2 groups were compared,and pharmacoeconomic analysis was conducted. RESULTS:Negative conversion rates of HCV-RNA in group A in 4,12,24,36,48 weeks were 55.00%,71.67%, 63.33%,61.67% and 65.00%,group B were 64.81%,66.67%,62.96%,55.56% and 61.11%,respectively,there were no signifi-cant differences between 2 groups (P>0.05);after treatment,normalization rate of ALT in group A was 95.23%,group B was 96.10%,there was no significant difference between 2 groups(P>0.05);and there were no significant differences in the incidence of ADR between 2 groups (P>0.05),so cost-minimization analysis was used to evaluate pharmacoeconomics. Therapy cost in group A was 13 216.56 yuan,group B was 7 929.60 yuan,group B was lower to group A;sensitivity analysis received the same results. CON-CLUSIONS:Interferonα2b is more economical thanα1b in the treatment of chronic hepatitis C.

Objective:To investigate the effects of TGF-β1 on T lymphocytes of BALB/c mice infected with Echinococcus granulosus( E.granulosus ) in vitro.Methods: The inhibitor group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and SB525334.The control group:the spleen cells of BALB/c mouse were co-cultured with E.granulosus and PBS.The blank group:the spleen cells of BALB/c mouse were co-cultured with RPMI-1640 medium and SB525334.The lymphocytes were collected at 48 h post-infection.The T lymphocyte subsets, the number of CD4+CD25+T cells, the number of NK cells, and the expression of NKG2D receptor were detected by flow cytometry.The NK cell activity was determined with the lactate dehydrogenase leakage assay(LDH).Results:The inhibition the TGF-β1 receptors resulted in the increase of in the number of CD4+T cells,the decrease in the number of CD8+T cells,the increase of in the ratio CD4+/CD8+T cells,the decrease of in the number of CD4+CD25+T cells,the increase in the expression of the NKG2D receptors,the increase in the lysis rate of Yac-1 cells by NK cells,and a positive cor-relation between the expression of activity receptor NKG2D and killing activity of NK, which were mediated by E.granulosus.Conclusion: The inhibition of TGF-β1 receptors can enhance the immune response of T lymphocytes against E.granulosus infection in vitro.

Objective:High-flow nasal cannula (HFNC) is a new method to treat adult respiratory distress. This study aims to explore the early predictors of the inefficiency of HFNC in patients with respiratory distress.Methods:A total of 162 patients with respiratory distress were treated with HFNC oxygen therapy in the Department of Respiratory and Critical Care Medicine, Emergency and Critical Care Medicine. The age, sex, weight, history, diagnosis, vital signs, blood oxygen saturation/inhaled oxygen concentration (SpO 2/FiO 2, SF) ratio, and modified respiratory distress score (mRDAI) , duration of HFNC oxygen therapy, replacement of advanced oxygen therapy support time, adverse reactions and other data were extracted from the medical electronic medical records. Results:A total of 154 valid samples were included, with a median age of 55 (inter-quartile range 47-72), including 59 patients (38.3%) with acute bronchiolitis, 64 patients (41.6%) with bacterial pneumonia, 31 patients (20.1%) with atypical or viral pneumonia; 129 patients (83.8%) with effective HFNC oxygen therapy and 25 patients (16.2%) with ineffective HFNC oxygen therapy. SF ratio in ineffective HFNC oxygen therapy patients at admission was lower, pH value was lower, partial pressure of carbon dioxide was higher ( P values were 0.008, 0.012, 0.001). RR, mRDAI score and SF ratio in the first hour of effective HFNC oxygen therapy patients improved significantly ( t values were 1.732, 9.783, 37.591, P<0.05 or 0.01). SF ratio in the first hour of HFNC was lower than 195 (area under curve 0.842, 95% CI 0.743-0.942, P<0.01). It was the critical value for the ineffectiveness of oxygen therapy. Conclusions:When patients with respiratory distress are supported by HFNC oxygen therapy, the lower initial fingertip oxygen saturation, higher blood gas arterial partial pressure of carbon dioxide and lower SF ratio are the early predictors of failure of HFNC oxygen therapy.

Objective To explore the clinical manifestations, imaging and pathology features, treatment, and prognosis of endocrine glands involved patients with IgG4-related disease ( IgG4-RD) . Methods Ten patients admitted in Peking Union Medical College Hospital from 1 January 2014 to 30 June 2018 diagnosed as IgG4-RD with endocrine glands involved were enrolled in this study. All the clinical data were collected and analyzed. Results Ten patients, 4 males and 6 females, median 55 years old at the onset were enrolled, five patients with single organ involvement ( 1 case involved in pituitary, and 4 cases involved in thyroid) , while another 5 patients with 2 and more organs involved. C-reactive protein or erythrocyte sedimentation rate was elevated in 8 patients, antinuclear antibodies were positive in 7, and serum total IgE was elevated in 6 patients. Nine patients were treated with oral glucocorticoids, among whom 3 patients were treated in combination with immunosuppressive agents or rituximab. After treatment, the clinical symptoms were alleviated and imaging was improved in all patients. Serum IgG levels were significantly decreased in all patients and normalized in 5 patients. Conclusion For the patients with multiple endocrine gland diseases and elevated IgG4 level, the possibility of IgG4-RD should be carefully considered and should be confirmed by tissue biopsy and histopathology.

Objective: To describe the clinical characteristics of pneumococcal infections and drug resistance of Streptococcus pneumoniae isolates from children's hospitals, which would provide reference for preventing and treating pneumococcal diseases. Methods: This was a prevalence survey. In this study, the age, specimen type, monthly distribution characteristics, and antimicrobial resistance of Streptococcus pneumoniae isolates from 9 children's hospitals in China were investigated between January 1, 2016 and December 31, 2016. The WHONET 5.6 software was used to analyze the antibiotic susceptibility of Streptococcus pneumoniae. The comparison of rates was performed by Chi-square test. Results: A total of 6 200 isolates of streptococcus pneumoniae were obtained, namely, 95.1% (5 876/6 177) from the respiratory tract specimens, 2.2% (136/6 177) from blood specimens and 0.4% (24/6 177) from cerebrospinal fluid specimens. The isolates were mainly from children older than 1 and younger than 5 years (54.7%, 3 381/6 185) . Most of strains (33.2%, 1 184/3 563) were isolated in November, December and January. Streptococcus pneumoniae isolates were completely sensitive to vancomycin (100.0%, 6 189/6 189) , linezolid (100.0%, 6 030/6 030) , moxifloxacin (100.0%, 3 064/3 064) , highly sensitive to levofloxacin (99.8%, 5 528/5 540), ertapenem (98.8%, 3 024/3 061) and lowly sensitive to erythromycin (1.7%, 102/6 016), clindamycin (3.7%, 116/3 136), and tetracycline (5%, 244/4 877), respectively. According to the parenteral susceptibility breakpoints for non-meningitis isolates, the sensitivity of Streptocococus pneumoniae to penicillin from children's hospital of Chongqing Medical University (49.3%, 892/1 809) was significantly lower than those of other hospitals (χ²=1 268.161, P<0.05) . Conclusions Streptococcus pneumoniae is mainly isolated from respiratory tract, from children older than 1 and younger than 5 years and during November to January in tertiary children's hospital of China. The Streptococcus pneumoniae from children is highly sensitive to vancomycin, linezolid, moxifloxacin, levofloxacin. There are also significant differences in the sensitivity of penicillin for Streptococcus pneumoniae from different hospitals.

Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.