AIM:To study the direct reprogramming method of mouse embryonic fibroblasts (MEFs) conver-ted into induced neural stem cells (iNSCs).METHODS:Sox2-infected MEFs were cultured in NSCs culture medium for 10 d.Subsequently , repeated suspension and adherent culture were performed for 3 times for the purification of iNSCs .The iNSCs were cultured in suspension medium .Real-time PCR was used to detect the expression of neural stem cell marker genes and pluripotent marker gene .In vivo, iNSCs were microinjected into the mouse cerebral cortex .Immunofluorescence was performed to detect the expression of neural stem cell , neuron, oligodendrocyte and astrocyte markers in vitro and vivo. RESULTS:A variety of neural stem cell marker gene expression was significantly increased in iNSCs detected by real -time PCR.Immunofluorescence confirmed that iNSCs expressed nestin and differentiated into neurons , oligodendrocytes and as-trocytes in vitro and vivo.CONCLUSION:Sox2 is sufficient to trigger the direct reprogramming from MEFs to iNSCs .iN-SCs have the ability of self-renew and 3 differentiation potentials in vivo and vitro.iNSCs are the suitable seed cells of SCI .

BACKGROUND:Poly(lactide-co-glycolide) (PLGA) scaffold is widely used in tissue engineering, but its poor cel adhesion ability and strong hydrophobicity limit its further development and application. OBJECTIVE: To study the biocompatibility of electrospun poly (lactide-co-glycolide)/polyethylene glycol (PLGA-PEG) nanofibrous scaffolds with mouse neural stem celsin vitro. METHODS:Neural stem cels were isolated from embryos of CD-1 mice at 15 embryonic days. Electrospinning was used to prepare PLGA and PLGA-PEG nanofibrous scaffolds. Scanning electron microscope was used for scanning observation of scaffolds. The 5th passage neural stem cels were seeded onto PLGA and PLGA-PEG scaffolds respectively, and culturedin vitro. RESULTS AND CONCLUSION: Interconnected porous network structure was observed in both two kinds of scaffolds under the scanning electron microscope. Fiber diameters and porosities of PLGA and PLGA-PEG scaffolds showed no significant differences (P > 0.05). Cel Counting Kit-8 detection showed neural stem cels grew wel on both two kinds of scaffolds and the absorbance value of two groups increased continuously with incubation time (1, 3, 5, 7, 9, 11 days). And there were statisticaly significant differences in the absorbance values between two groups at each time point (P < 0.05). Moreover, the cel adhesion rate was significantly higher in the PLGA-PEG group than in the PLGA group at 3, 6, 9 hours of culture (P < 0.05). Hoechst 33342 staining showed normal morphology and quality of the nuclei, and significantly more cels were observed in the PLGA-PEG group than the PLGA group (P < 0.05). Under the scanning electron microscope, compared with the PLGA scaffold, the PLGA-PEG scaffold was better for growth and matrix secretion of neural stem cels. In conclusion, PLGA-PEG nanofibrous scaffolds prepared by electrospinning are safe, non-toxic and suitable for neural stem cels growth with wel biocompatibility, appropriate aperture and porosity.

Objective To analyze the occurrence features of the active surveillance of adverse events following immu?nization (AEFI) in Tianjin, and evaluate the sensitivity of passive surveillance systems over the same period. Methods The layered eight vaccination clinics of four counties were selected as active surveillance points. The data of active surveillance AEFI were collected through telephone investigation in 2011-2013. The data of passive surveillance was collected through the national AEFI information management system. The descriptive methodology and chi square test were used. Results A total of 235 cases were collected through active surveillance in 2011-2013. The AEFI incidence rate was 718.70 per 100 000 doses,no difference in the incidence between three years(χ2=5.07). A total of 4164 cases were collected through passive sur?veillance. The incidence rate was 34.09 per 100 000 doses,the incidence increased year by year(χ2=572.02,P<0.05). In ac?tive surveillance cases, 95.32%was the common vaccine reaction, 3.83%was the rare vaccine reaction, no serious vaccinereaction. In passive surveillance cases, the common vaccine reaction, the rare vaccine reaction were 85.09%and 13.32%. The serious vaccine reaction was found in 44 cases. Conclusion The more slightly common vaccine reaction can be found by active surveillance. The sensitivity of Tianjin passive surveillance system is increasing year by year, which can meet the requirements for job. The classification and diagnostic criteria of the serious vaccine reaction need to be established.

With the development of bio-technological drugs, drug immunogenicity evaluation has become key factor of clarifying safety and efficacy of these drugs. It has become the focus to establish a stable and reliable evaluation system. Due to the advantages such as continuous real-time monitoring, surface plasmon resonance (SPR) technology has been widely used in bio-technological drugs immunogenicity assessments. Our study applied this technology to detect anti-drug antibody (ADA) of a recombinant human anti-rabies monoclonal antibody NM57 in the sera of 48 volunteers admitted in phase I clinical trials. This method could satisfy the basic requirements of detection of ADA.

Objective To analyze the correlation between CT characteristics and epidermal growth factor receptor (EGFR)mutations in pulmonary adenocarcinoma.Methods 82 patients (87 lesions)with pulmonary adenocarcinoma were retrospectively collected.All patients were underwent CT examination before operation,and EGFR gene were determined after operation.Results EGFR mutations were found in 44 of 87 lesions (50.57%).The EGFR mutations rate was 50.00% in females and 47.50% in males,there was no statistical difference between genders (P=0.821).The EGFR mutations rate was 46.55% in the right lung and 58.62% in the left lung,while no statistically significant difference was found (P=0.289).Among all the CT characteristics,the mutations rate was 63.89% in spiculated lesions and 60.71% in lesions with pleural indentation,the differences was statistically significant (P<0.05).The mutations rate was 59.25% in lesions containing solid component and 36.36% in pure ground glass opacity lesions,the difference was statistically significant (P=0.011). There were no statistically differences in lobulation,cavitation and lymphadenectasis (P>0.05).The pleural indentation was the highest in sensitivity (77.27%)and negative predictive value (67.74%).The spiculation was the highest in specificity (69.77%)and positive predictive value (63.89%).Conclusion Among all the CT characteristics,pleural indentation,spiculation and the lesion containing solid component are prone to EGFR mutations.

ObjectiveTo determine the efficacy of constraint-induced movement therapy(CIMT) on different severity of the motor deficit of upper extremity after stroke.Methods27 stroke patients who had upper-limb hemiparesis and learned nonuse were allocated either to the moderate group(n=12) or the severe group(n=15) according to severity of initial motor deficit of upper extremity by Upper Extremity Function Test(UEFT).The two group patients were given CIMT,involving restriction of movement of the intact upper extremity by placing it in a sling for 90% of waking hours for 12 days and training(by repetitive practice,shaping) of the more affected extremity for 6 hours on the 10 weekdays during that period.Outcome were measured with UEFT.ResultsThere was no significant difference in treatment gain on the UEFT between the moderate patients and the severe patients(P>0.05).However,the Effect Size for the severe patients(ES=2.2) was larger than for the moderate patients(ES=1.8) at the quality of movement(UEFT).ConclusionCIMT is an effective rehabilitation technique for different severity of the motor deficit of upper extremity after stroke,especially in severe stroke patients.

Objective To study the effect of rhBNP on serum chemerin and IL-37 levels in acute myocardial infarction (AMI) patients undergoing emergency PCI.Methods Eighty AMI patients who underwent emergency PCI were randomly divided into cortrol group (n =40) and rhBNP treatment group (n=40).The patients in control group were treated with conventional drugs and those in rhBNP treatment group were treated with intravenous rhBNP.Their serum chemerin and IL-37 levels were measured by ELISA.Their LVEDD and LVEF were compared.Results The serum level of chemerin was significantly lower while that of IL-37 was significantly higher in two groups at 72 h and on day 7 after PCI than before PCI (P＜0.05).The serum level of chemerin was significantly lower while that of IL-37 was significantly higher in rhBNP group than in control group at 72 h and on day 7 after PCI (P＜0.05).The LVEDD was significantly shorter while the LVEF was significantly higher in two groups on day 7 and month 1 after PCI than before PCI (P＜0.05).The LVEDD was significantly shorter in rhBNP group than in control group on day 7 and month 1 after PCI (P＜0.05).Conclusion rhBNP can effectively reduce the serum chemerin level,increase the serum IL-37 level,and improve the cardiac function in AMI patients following emergency PCI.The effect of rhBNP is better than that of conventional drugs in AMI patients after emergency PCI.

Objective To investigate the effect of C1q/tumor necrosis factor-related protein 13 protein(CTRP13)on the autophagy function of primary rat liver sinusoidal endothelial cells(rLSECs)induced by high glucose through AMP-activated protein kinase/mammalian target of rapamycin complex(AMPK/mTOR)pathway.Methods After isolation,identification and culture,original rat liver sinusoid endothelial cells(rrLSECs)were divided into normal control(NC)group,high glucose(HG)group,HG +LV-CTRP13 group,HG+ lentiviral empty vector(LV-Con)group(HG+LV-Con).CTRP13 lentivirus over expression vector(LV-CTRP13)and lentivirus empty vector(LV-Con)were constructed and transfected into rrLSECs.According to the intervention methods of AMPK inhibitor Compound C,mTOR inhibitor Torin1 and autophagy inhibitor 3MA,the transfected cell were divided into normal control(NC)group,high glucose(HG)group,HG+LV-CTRP13 group,HG+lentiviral empty vector(LV-Con)group(HG+ LV-Con).qRT-PCR and western blot were used to detect the mRNA and protein expression levels of CTRP13,autophagy related protein Beclin1,human microtubule-associated protein light chain 3II(LC3II),human plasma membrane membrane vesicle association proteins(PLVAP)and p-AMPK and p-MTOR in rat rLSECs of each group.Results Compared with NC group,the number of autophagosome was decreased in HG and HG+LV-CTRP13 group(P＜0.05).Compared with HG group,the number of autophagosome bodies was increased in HG +LV-CTRP13 group(P＜0.05).The CTRP13 mRNA and protein expression was higher in NC and HG + LV-CTRP13 groups than in HG and HG + LV-Con groups(P＜0.05).In HG+LC-CTRP13 group,Beclin1,LC3II,p-AMPK,and AMPK mRNA,Beclin1,LC3II/LC3I protein expression were higher than HG and HG + LV-Con group(P＜0.05),PLVAP,p-mTOR,mTOR mRNA,and PLVAP protein expression were lower than HG and HG+LV-Con group(P＜0.05).Comparison with HG + LV-CTRP13,p-mTOR protein expression in HG+LV-CTRP13+Compound C group increased(P＜0.05),while expressions of CTRP13,Beclin1 and LC3II/LC3I protein decreased(P＜0.05);the protein expressions of p-AMPK,Beclin1 and LC3II/LC3I were increased in HG+LV+ CTRP13+Torin1 group(P＜0.05),while CTRP13 and p-mTOR protein expression was decreased(P＜0.05);protein expressions of p-AMPK,p-mTOR and LC3II/LC3I were higher in HG+LV-CTRP13 + 3MA group(P＜0.05),while LC3II/LC3I protein expression was lower(P＜0.05).Conclusion CTRP13 overexpression activates AMPK/mTOR-autophagy signaling pathway,which may play a protective role in the function of rLSECs anddelay liver sinusoid capillarization.

Objective:To investigate the clinical effect of Infix combined with hollow screws for the treatment of pelvic injuries with pubic symphysis separation in middle-aged and elderly patients.Methods:Data of 8 middle-aged and elderly patients with pelvic injuries due to pubic symphysis separation undergone treatment from January 2017 to December 2020 were retrospectively analyzed.Results:The average operating time of 8 patients was (46.0±6.2)min(range: 40-62min); the average intraoperative blood loss was (32.0±5.6)ml(range: 25-50 ml); the average length of incisions at the iliac screw was(2.6±0.4)cm(range: 2.0-3.5 cm); the average length of incisions at the hollow screw was (1.1±0.3)cm(range: 0.8-1.5 cm); and the average times of fluoroscopy were (36.0±6.0)times(range: 28-52 times). Postoperative X-ray and CT examinations showed that the reduction of the pubic symphysis was good, the inserted iliac screws and cannulated screws were positioned accurately, and the incision healed well.Based on Matta's criteria, postoperative radiological outcomes were evaluated, with 7 cases rated as excellent and 1 as good, giving an excellent to good rate of 100%(8/8). The average followed up time for all 8 patients was (15.0±4.2)months(range: 6-24 months). Pelvic X-ray and CT examinations at the last follow-up showed that the fractures healed well and the pubic symphysis reduction did not fail.Infix and cannulated screws in the pubic symphysis were removed 10-12 weeks after surgery[average: (10.5±0.5)weeks]. According to the Majeed Pelvic Score, 5 cases were rated as excellent, 2 cases as good and 1 as fair, with an excellent to good rate of 87.5%(7/8). One patient had symptoms related to the lateral femoral cutaneous nerve that disappeared after 3 months.One patient developed deep venous thrombosis after surgery, and the filter was placed and removed 10 weeks later.Conclusions:Using Infix plus cannulated screws for the treatment of pelvic injuries in middle-aged and elderly patients with pubic symphysis separation has the advantages of limited trauma and intraoperative blood loss, good fixation and few complications.

Objective:To analyze the differences of ocular surface bacteria between patients with type 2 diabetes and healthy subjects, and to explore the relationship between ocular surface bacteria and type 2 diabetic keratopathy.Methods:Fifty subjects were selected and divided into type 2 diabetes group and healthy control group. Bacterial culture of conjunctival sac secretion was conducted and 16S rRNA sequencing was performed to analyze the difference in the composition of ocular surface bacteria between patients with type 2 diabetes and healthy subjects.Results:There were significant differences in the diversity of ocular surface bacteria between patients with type 2 diabetes and healthy subjects. The species of bacillus brevis and paenibacillus in type 2 diabetes group were significantly higher than those of the healthy control group, while the species of aquatic bacteria were lower.Conclusion:The differences in the composition of ocular surface bacteria may be related to type 2 diabetes mellitus.