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.

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 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.

Objective:To investigate the clinical effect of using combined lumbar pelvis fixation device in the treatment of old vertical unstable fracture dislocation of posterior pelvic ring.Methods:Data of 7 patients with old vertical unstable fracture dislocation of posterior pelvic ring admitted and followed up from January 2017 to April 2020 were retrospectively analyzed, including 4 males and 3 females with an average age of 42.4 years old (range, 22-73 years old). There were 3 cases of traffic injury, 3 cases of falling injury and 1 case of tumble injury. According to Tile classification for pelvic fractures, there were 5 cases of type C1, 1 case of type C2 and 1 case of type C3. The average time from fracture to surgery was 5.4 weeks (range, 3-10 weeks). Among the 7 patients, 4 patients' posterior ring fractures were fixed by combined lumbar pelvic triangle fixation, and 3 patients' posterior ring fracture were fixed by combined lumbar pelvic fixation. 4 patients' anterior ring injury were not treated, 2 patients' anterior ring injury were treated by closed cannulated screw and internal fixation, and 1 patient's anterior ring injury was fixed by INFIX and cannulated screw. Every patient's operating time, intraoperative blood loss, length of incision and times of X-ray fluoroscopy were recorded. Pelvic X-ray and CT scan were taken postoperatively to observe the condition of reduction and screw position. Postoperative fracture reduction quality was assessed by Matta radiological criteria and Majeed criteria was used at the final follow-ups to evaluate the degree of functional recovery after pelvic fracture.Results:The average operating time of 7 patients was 143 min (range, 96-205 min); the intraoperative average blood loss was 579 ml (range, 300-1 650 ml); the average length of incisions was 12.9 cm (range, 9-15 cm) and the average time of X-ray fluoroscopy was 27 times (range, 15-52 times). Postoperative X-ray and CT scan showed that the displacements of the posterior rings were reset well and all the hollow screws were located accurately and firmly. Postoperative radiation quality was evaluated according to Matta radiological criteria, and there were 4 cases of excellent, 2 cases of good and 1 case of fair, with an excellent and good rate of 85.7% (6/7). Seven patients had good fracture union. The average followed up time for all 7 patients was 12 months (range, 6-16 months). At the last follow-up, imaging examination showed good reduction of the sacroiliac joint, and the reduction of anterior and posterior rings were not lost. The healing time was 14.2 weeks (range, 12-20 weeks). Majeed score: postoperative 4.90±6.64 points (range, 48-58 points), postoperative 3 months 71.40±7.32 points (range, 67-75 points), postoperative 6 months 84.90±8.14 points (range, 68-96 points), the difference was statistically significant ( F=0.614, P=0.004). Majeed score 6 months after operation showed that 5 cases were excellent, 1 case was good and 1 case was fair, and the excellent and good rate was 85.7% (6/7). Conclusion:Using combined lumbar pelvis fixation device in the treatment of old vertical unstable fracture dislocation of posterior pelvic ring has good reduction quality, high fixed strength and good postoperative effect.

Objective:To explore the changes of corneal neuropathy and ocular surface flora in patients with type 2 diabetes, and the correlation between corneal neuropathy and ocular surface flora changes.Methods:According to the results of fundus fluorescein angiography, 65 patients with type 2 diabetes (130 eyes in total) who were treated with insulin alone and without comorbid retinopathy in the Department of Endocrinology, Henan People′s Hospital from March to June 2019 were selected. Sixty-five age-matched normal glucose tolerance subjects (130 eyes in total) were also enrolled. Confocal microscopy was performed and conjunctival sac secretions were collected. Through 16S rRNA analysis and sequencing and PICRUSt bacterial gene function prediction, the relationship between patients with type 2 diabetes and the difference in the degree of corneal neuropathy and the composition of ocular surface flora and function prediction in healthy people were explored, and the correlation between corneal neuropathy and ocular surface flora was analyzed as well.Results:Compared with healthy people, patients with type 2 diabetes have more severe corneal neuropathy, and it was related to the changes in ocular surface flora. Brevibacillus and paenibacillus were significantly positively correlated with the degree of corneal neuropathy ( P<0.05), and enhydrobacter and proteobacteria were significantly negatively correlated with the degree of corneal neuropathy ( P<0.05). PICRUSt analysis showed that the degree of enrichment of metabolism-related genes in the ocular surface flora of patients with type 2 diabetes was significantly changed compared with healthy people. Conclusion:Patients with type 2 diabetes present with more severe corneal neuropathy. The diversity of ocular surface flora and metabolic function have significant changes. The degree of severity corneal neuropathy is related to brevibacillus, paenibacillus, enhydrobacter, and proteobacteria.

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.