Objective To study the transduction efficiency of expressing human neprilysin by using a lentiviral vector (Lenti-NEP) in mouse embryonic neural stem cells (NSC) in vitro.Methods Primary NSC were harvested from C57BL/6J pregnant mouse at embryonic day 11.5 and transducted with LentiNEP.Immunofluorescent stainingand Western blot were performed to detect NEP protein expression in NSC.Degradation of amyloid beta 1-40 (Aβ1-40) by NEP protein transduced with Lenti-NEP in NSC was analyzed using ELISA and HPLC.Results Over 90％ NSC were successfully transduced with Lenti-NEP via observation of fused protein green fluorescent protein under the microscopy.Expressions of NEP transduced with Lenti-NEP in NSC and of the markers of NSC (nestin) and neuron (MAP2).The enzyme activity of 2.5 μg (21.00 ± 2.51) and 1.0 μg (15.00 ± 0.54) NEP on degrading Aβ1-40 was shown to improve significantly compared to 0.5 μg NEP(8.00 ±0.81,t =40.4 and 12.7,respectively,both P ＜0.01).The activity of NEP was inhibited in the presence of 50 μmol/L phosphoramidon (0.5 pg:0.08 ±0.01 ;1.0 μg:0.04 ±0.01 ;2.5 μg:0.05 ±0.01,t =17.2,51.3 and 14.1,respectively,all P ＜0.01).The hydrolytic cleavage on degrading Aβ1-40 by NEP was 11.4％,28.4％ and 93.7％ with incubation for 1 h,4 h and 12 h,respectively.Conclusions Lentiviral vector successfully delivers NEP gene to NSC in vitro.Targeting on NEP and NSC may provide potential therapeutic tool for Alzheimer' s disease.

Objective To investigate the therapeutic effect of exosomes extracted from human adipose-derived mesenchymal stem cells(hAMSCs) on traumatic brain injury (TBI) and its possible mechanism.Methods Mesenchymal stem cells(MSCs) were isolated from healthy human adipose tissue and the exosomes were extracted by ultrafiltration.Rats were divided into four groups: sham group, PBS control group, MSCs treatment group and exosomes treatment group.24 h After TBI, the treatment group was locally injected along the lesion area, 30 μL of PBS, 2×105 MSC, 25 μg protein of exosomes respectively, the total volume was 30 μL.We performed the Modified Neurological Severity Score(mNSS) and the forelimb Foot-Fault Test in all rats before injury and at 1, 3, 7, 10, 13, 16, 21 and 30 days after TBI.The rats were sacrificed at 3 and 7 days after TBI respectively,total RNA was extracted from rat brain tissue.The expression of TNF-α and IL-1β were detected by quantitative PCR.The rats were also killed at 30 days after TBI for testing the neuronal apoptosis in lesion area by tunel-neun double imm-unofluorescence.Results Exosomes treatment significantly promotes the recovery of neurological deficits caused by TBI,and the therapeutic effect is similar to MSCs, its possible mechanism may be the inhibition of the acute inflammation and the reducing of the neurons apoptosis after TBI.Conclusions Exosomes extracted from human adipose-derived mesenchymal stem cellshas promoted neurological functionrecovery after traumatic brain injury, which will provide a new and safer TBI treatment for clinical practice.

Objective To retrospectively analyze the prognostic factors for locoregionally recurrent early?stage extranodal nasal?type natural killer/T?cell lymphoma ( NKTCL) . Methods A total of 56 patients with early?stage extranodal nasal?type NKTCL, who had locoregional recurrence after initial treatment and then received salvage treatment from 1995 to 2014, were enrolled as subjects. The effects of salvage treatment on the overall survival ( OS) rate were analyzed after initial treatment and recurrence. Univariate and multivariate prognostic analyses were performed on the OS rate after recurrence. Results The median follow?up time was 35. 9 months after initial treatment and 14. 8 months after recurrence. The 3?year OS rate was 73% after initial treatment and 58% after recurrence. Compared with chemotherapy alone, radiotherapy?containing salvage treatment significantly improved the OS rates after initial treatment and recurrence ( P=0. 040, 0. 009 ) , and re?irradiation also significantly improved the OS rates after initial treatment and recurrence (P=0. 018, 0. 019). Most (84%) of the acute and late adverse reactions after re?irradiation were grade 1?2 ones. The univariate and multivariate analyses showed that the Karnofsky Performance Status score, radiotherapy in initial treatment, and radiotherapy in salvage treatment were influencing factors for the OS rate after recurrence. Conclusions Radiotherapy achieves improved survival and tolerable toxicities, making it indispensable in the treatment of locoregionally recurrent extranodal nasal?type NKTCL.

Objective To analyze γ?secretase gene mutations in a pedigree with acne inversa. Methods Clinical data were collected from a pedigree with acne inversa, which contained 30 members spanning 4 generations. Of these members, 12 were affected by acne inversa, and 9 of the affected members were alive. Peripheral blood DNA was obtained from the proband, his seven relatives (including 4 affected and 3 unaffected members), and 100 unrelatedhealthy human controls. PCR was performed to amplify all the coding exons and their flanking sequences of the NCSTN, PSEN1, PSENEN, Aph1 genes followed by DNA sequencing. Results A heterozygous insertion mutation (c.229_230insCACC)of the PSENEN gene, which led to translational frameshifting and resulted in dysfunciton of the PSENEN protein, was detected in all the 5 patients, but not in unaffected members or healthy controls. Conclusion There is a novel heterozygous insertion mutation c.229_230insCACC in the PSENEN gene, which may be the molecular basis of acne inversa in this family.

AIM:To investigate the efficiency of 2-methoxyestradiol (2-ME) as radiosensitizing agent for the treatment of lung cancer cells. METHODS:Cell line A549 and GLC-82 originated from human non-small cell lung cancer were cultured in vitro. Study group (2-ME in different concentrations) and control group without 2-ME were set up. Cell proliferation was measured by MTT assay that lung cancer cells were treated with 2-ME for 24 h,then the cells were exposed from 0 to 8Gy radiation,and the survival fraction was determined by clone forming test. Flow cytometry was used to measure the effects of 2-ME on cell cycle distribution. RESULTS:MTT assay showed minimum effective concentration value was 0.15625×10~(-6) mol/L in GLC-82 and 1.25×10~(-6) mol/L in A549 cells. Compared to control group,exposed GLC-82 cells or A549 cells to minimum effective concentration of 2-ME for 24 h before irradiation resulted in an enhancement of radiation. The protection enhancement factor was 1.98 and 2.06 in GLC-82 and A549 cells,respectively. Flow cytometry analysis of cell cycle progression demonstrated G_2/M phase arrest in both cells in a dose dependent manner. No obvious change of CDK2 activity in both GLC-82 cells and A549 cells was observed. CONCLUSION:2-ME enhances radiosensitivity by G_2/M phase arrest in the cell cycle.

Objective:To investigate the application value of laparoscopic duodenum-preserving pancreatic head resection (LDPPHR).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 25 patients undergoing LDPPHR in the Second Hospital of Hebei Medical University from November 2016 to November 2020 were collected. There were 7 males and 18 females, aged from 14 to 66 years, with a median age of 29 years. All the 25 patients underwent LDPPHR. Observation indicators: (1) surgical situations; (2) postoperative histopathological examination; (3) follow-up. Follow-up was conducted using outpatient examination or telephone interview to detect the recovery of patients up to March 2021. Measurement data with skewed distribution were represented as M (range) and count data were descripted as absolute numbers. Results:(1) Surgical situations: all the 25 patients underwent LDPPHR successfully, including 23 cases undergoing total pancreatic head resection and 2 cases undergoing subtotal pancreatic head resection. The operation time and volume of intraoperative blood loss of 25 patients were 310 minutes (range, 207 to 540 minutes) and 200 mL (range, 50 to 800 mL), respectively. Of the 25 patients, 1 case was infused with 4 U of red blood cells and 400 mL of plasma, 1 case was infused with 500 mL of plasma, 1 case was infused with 600 mL of plasma and the remaining 22 cases were not infused with red blood cells or plasma. Of the 25 patients, 3 cases with pancreatic fistula of class B were discharged after drainage, 4 cases had biliary fistula including 2 cases undergoing symptoms disappeared after implantation of common bile duct stent by endoscopic retrograde cholangiopancreatography, 1 case recovering well with drainage, 1 case with postoperative perihepatic effusion undergoing symptoms disappeared after the treatment of drainage and common bile duct stent implantation, and the remaining 18 cases had no complications. The duration of postoperative hospital stay was 17 days (range, 9 to 27 days) of the 25 patients. (2) Postoperative histopathological examination: the tumor volume of the 25 patients was 6.0 cm×5.0 cm×2.0 cm (range, 1.0 cm×2.0 cm×1.5 cm to 10.0 cm×9.0 cm×8.0 cm). Results of the postoperative histopathological examination showed that there were 12 cases with pancreatic solid pseudopaillary neoplasm, 4 cases with intraductal papillary mucinous neoplasm, 3 cases with serous cystadenoma, 2 cases with mucinous cystadenoma, 1 case with neuroendocrine neoplasm, 1 case with pancreatic true cyst, 1 case with cholesterol crystals combined with calcification in the center of pancreatic nodules and 1 case with cavernous hemangioma of pancreas. (3) Follow-up: all the 25 patients were followed up for 4 months to 48 months, with a median follow-up time of 27 months. During the follow-up, 1 case of the 25 patients with postoperative diabetes controlled blood glucose in the normal range after regular injection of insulin, 1 case with fatty diarrhea had symptoms improved after oral supplement of pancreatic enzyme preparation, 1 case with preoperative intermittent dizziness, weakness of both lower limbs and hypoglycemia had the level of blood glucose returned to normal without any special treatment after operation, and the remaining 23 cases had no metabolic complications. None of the 25 patients had tumor malignant transformation, recurrence or death. No long-term complications such as delayed gastric emptying, bile duct stones or stricture occurred to the 25 patients.Conclusion:LDPPHR is safe and feasible for the treatment of benign or low-grade malignant tumors of the pancreatic head, with the advantage of preserving the integrity of digestive tract.

Objective: To demonstrate the effect of structured and autonomous sports activities on children s basic motor skills in a preset outdoor field for a certain time and frequency, and to provide a reference for improving children s development of basic motor skills. Methods: Selected 141 typical developing children from 4 public kindergartens in Nanning, and randomly divided them into experimental group(71) and control group(70). The experimental group participated in physical activities for 10 weeks (3 times a week, 1 hour each time) in the preset outdoor field (each physical activity was composed of 30 minutes of autonomous physical activity and 30 minutes of structured physical activity), while the control group had no intervention of site preset and specially designed activities. Before and after the experiment, 9 basic motor skills (including 3 fine motor skills and 6 gross motor skills) were tested. Results: There were significant differences in double hand throwing, single leg balance (left), single leg balance (right), swing bridge balance and beam walking before and after intervention in experimental group(t=7.82,4.25,9.69,-10.01,-28.28,P<0.05), but there was no statistical significance in the improvement of heel toe coordinated walking(t=-0.60,P>0.05). There was no significant improvement in coin throwing (right hand), coin throwing (left hand) and square stacking(t=0.91,1.03,0.42,P>0.05), while in the control group, except for the balance of swinging bridge(t=4.27,P<0.05) and heel and toe walking(t=3.96,P<0.05), the improvement of other basic motor skills was not statistically significant(t=1.21, 0.74, 0.31, 0.97,P>0.05). Conclusion The quality of sports environment has a potential impact on the basic motor skills of preschool children. The basic motor skills of the pre-school children can be greatly improved by conducting structural and autonomous physical activities with a certain time and frequency.

Objective To explore the morphometric and functional alterations of amygdale and hippocampus in patients with depression by anatomical and functional MRI, and try to reveal the pattern and pathogenesis of the changes in depression. Methods Sixty patients (divided equally into mild, moderate and major groups according to patient′s scores of HAMD) and 20 healthy control groups were scanned using T1WI and fMRI. The outlines of hippocampus and amygdale were drawn manually by observer and the volumes were calculated and normalized subsequently. Functional MRI was processed using SPM5 and individual activation map was got subsequently. Dunnett-t test and Pearson correlation analysis were separately used to analyze the morphometric and functional changes and the correlations between cerebral changes and clinical severity. Results The hippocampal volumes of control groups were 2296±202 left for left side and 2283±199 for right side. The hippocampal volumes of depressive patients were smaller than those of control groups, especially for the major group (left 1978±176,Dunnett-t=-10.0,P<0.01,right 1981±171,Dunnett-t=-9.2,P<0.01). The moderate group showed moderate reduced volume(left 2127±180,Dunnett-t=-3.0,P<0.05,right 2135±183,Dunnett-t=-3.0,P<0.05), while the mild ones showed slightly decreased volume (left 2207±189,Dunnett-t=-1.4,P>0.05,right 2210±191,Dunnett-t=-1.6,P>0.05). The amygdale′s volumes of control groups was 1762±185,the right was 1749±182, while those in patient group reduced along with the patient′s condition, i.e., the mild groups (left 1992±200,Dunnett-t=4.8,P<0.01,right 1989±191, Dunnett-t=5.0,P<0.001), the moderate groups (left 1889±192, Dunnett-t=2.8,P<0.05,right 1896±195,Dunnett-t=2.8,P<0.05), and the major groups (left 1539±178,Dunnett-t=-6.8,P<0.01,right 1543±180,Dunnett-t=-7.0,P<0.01).For fMRI study, patient group demonstrated more activation of the amygdale and hippocampus under the stimulations of negative images than controls. Furthermore, the strengthens of activation decreased along with the patient′s condition, i.e., the major ones showed the weakest activation among the patients, though it was higher than that of control group. In patient group, both the volumes and activations of hippocampus and amygdale showed significant negative correlations with HAMD scores(r=-0.80--0.90,P＜0.05). Conclusion The hippocampal volumes of depressive patients reduced, which may be due to the change of the amygdale, and the amygdale′s volumes were changed along with the patient′s condition. There were more activation in the amygdale and hippocampus of depressive patients under the stimulations of negative images.

Objective To evaluate the clinical result of Hering's law in the correction of asymmetric bilateral ptosis.Methods From January 2014 to July 2016,165 cases divided into 2 groups with asymmetric bilateral ptosis in mild and moderate ptosis were corrected with aponeurosis advancement type correction:A group of 15 cases,in which the surgery does not consider the influence of Hering's law,the first correction of mild side,and then corrected the heavier side;B group of 150 cases,in which the Hering's law was applied,the first correction of severe side so that the margin reached the edge of the cornea is tangent or low 0.5-1.0 mm,and then corrected mild side.That the difference of symmetry between the two sides of the palpebral fissure was less than 0.5 mm,and the follow-up was more than 3 months.The Pearson chi square test was used to compare the differences between the two groups.Results In this group,165 cases were followed up for 3-24 months,10 cases (66.67％) was satisfactory in group A (15 cases);145 cases (96.67％) was satisfactory in group B (150 cases);After 3 months,all the failure cases got satisfactory result by using the Hering's law in corrective surgery again.The P value of chi square test between groups was less than 0.05.Conclusions The application of Hering's law in the correction of asymmetry bilateral ptosis is more satisfactory.

Objective To explore the impact of WeChat-assisted follow-up on self-administer medication capability of stroke survivors.Methods Seventy patients were recruited from a tertiary hospital and randomly divided into intervention group (n=35) and control group (n=35).Patients in intervention group were given WeChat-assisted follow-up including health education program and behavior guidance,and patients in control group received regular discharge care.The effects of WeChat-assisted follow-up and regular care on self-administer medication ability,cognitive and behavior level were assessed.Results After three months' follow-up,the scores of self-administer medication capability (38 (32,42)),experience (25 (20,28) and the integral level (87 (71,92)) in intervention group were significantly higher than those in control group (capability 25(16,38),experience 21 (8.75,27),total score 68 (38,87)) (Z=-2.511,-2.033,-2.209,P＜0.05).The self-administer medication experience,cognitive,capability and total scores increased significantly in intervention group after intervention(Z=-4.525,-4.610,-3.806,-4.718,all P＜ 0.01),while only the cognitive level increased slightly in control group (18 (10,24) vs 13 (11,18),Z =-1.794,P=0.073).Conclusions Follow-up intervention based on WeChat platform can improve the stroke patients' self-administration medication capability,however,the long-term effects on cognitive level and further reform need to be strengthened.