AIM: To investigate the effect of nerve growth factor (NGF) on the proliferation of neural stem cell in adult rats with focal cerebral ischemia. METHODS: The experiment was conducted in the Center of Science and Experiment, Jinzhou Medical College from January to July 2006. Totally 72 healthy SD rats were randomly divided into sham-operation group, model group and NGF treatment group with 24 rats in each group. Focal cerebral ischemia models were prepared by Logna et al modified thread occlusion method, and the functional evaluation was performed after the animals were awake for 2 hours to select the rats whose nervous function reached grade 2 or above. The sham-operation group was treated like the model group except thread occlusion. 1 000 ?g/kg NGF was administered in the NGF treatment group immediately after ischemia, once a day. After the animals were executed at days 1, 3, 7 and 14 days, immunohistochemical and double immunofluorescence labeling were adopted to observe the effect of NGF on the expression of nestin and the cell type after ischemia. RESULTS: All 72 rats were involved in the result analysis. ①Round or ellipse nestin positive cells were found in both the model group and NGF treatment group. The number of nestin positive cells in the NGF treatment group was remarkably more than in model group except 1 day after ischemia, moreover, the number of the cells in the two groups was more than in the sham operation group [model group: (3.47?0.5), (5.13?1.14), (13.95?3.56), (8.97?2.08); NGF group: (3.81?0.66), (9.88?2.08), (19.87?3.86), (26.17?2.90); sham-operation group: 0, P

Objective This study aims to examine the development of the posterior lateral line of the zebrafish and establish ant model to study the process of hair cell differentiation and regeneration .Methods We observed the posterior lateral line system formation by DAPI immunohistochemistry and whole mount in situ hybridization .We further evaluated hair cells differentiation within neuromast by using Transgenic Tg (Brn3c:mGFP) zebrafish and stained the functional hair cells by the mechanotransduction marker FM 1 -43FX .We labelled proliferating cells in primordium and neuromast by addition of BrdU to the system water .Results The posterior lateral line primordium originated from a sensory placode and started its journey at around 20 hours post fertilization to migrate along the horizontal myoseptum to the tail -tip with a constant speed (1 .7somite/hour) .The primordium depositd five or six neuromasts spaced along the body ,and two or three terminal neuromasts at the tail -tip at 48 hours post fertiliza-tion .At 3 ,5 and 7 days post fertilization ,zebrafish contained 5 .68 ± 1 .46 ,10 .1 ± 0 .99 ,and 12 .45 ± 1 .32 hair cells per neuromast ,respectively .Furthermore ,the average number of FM1-43FX stained hair cells within each neuro-mast were 3 .68 ± 1 .11 ,8 .18 ± 1 .86 ,and 10 .22 ± 1 .24 ,respectively .Conclusion We establish the development model of hair cells in zebrafish lateral line neuromast and suggest that 3 to 7 days post fertilization is an important period for lateral line neuromast differentiation .This study would be useful for underlying the mechanisms of hair cell differentiation and regeneration .

Objective To obtain an easy and high efficient method for gene transfer to cochlear hair cells ,by comparing three mediating green fluorescent protein (GFP) methods (electroporation ,adenovector and lentivirus vector) .Methods Cochlear sensory epithelium was dissected from anaesthetized P2 mice .Sensory epithelia were transferred onto poly -L -lysine treated cover slides and cultured overnight .Gene transfer was performed by elec‐troporation in medium containing pGPHI/GFP/Neo plasmid or by incubation with diluted recombined adenovirus/lentivirus vector .After 48 hours ,green fluorescence was checked under fluorescence microscope .To confirm the ef‐ficiency of exogenous gene transfer ,real-time PCR was performed using specific primers .Results The transfec‐tion efficiencies of electroporation and lentivirus vector mediated gene transfer were very low .Both immunofluores‐cence and real - time PCR results showed that the transfection efficiency of adenovirus mediated GFP and Bmi1 transfer were relatively higher .The proportion of GFP positive cells in outer hair cells and inner hair cells of middle turn were 90 .0% ± 4 .1% and 5% ± 0 .4% ,respectively .Conclusion Adenovector is more efficient for exogenous gene transfer to cochlear hair cells ,thus adenovector is a good carrier for gene transfer to cochlear hair cells .

Objective By investigating the actualities of career development potential of eight-year program medical students, to explore a direction of training mode innovation aiming to improve career development potential. Methods Eight-year program medical graduates, their tutors, and di-rectors of their clinical department in a university hospital were the subjects of the current study. Questionnaire method was applied to evaluate professional competence including clinical skills, re-search capabilities and integrative ability . Anonymous questionnaires were dispensed to eight-year program medical graduates for self-evaluation. Distinct anonymous questionnaires were dispensed to tutors directors for evaluation. Traditional doctoral graduates (professional degree or academic degree) were set as standard for comparison of specific ability of eight-year program medical students. Results 74 out of 85 (87%) questionnaires for Eight-year program medical graduates were available. 53 out of 63 (84%) questionnaires for tutors were available. 21 out of 26 (81%) questionnaires for directors of clinical departments were available. In clinical capabilities, according to 43 percent (32 persons) of the eight-year MD graduates' self-assessment and the evaluation of 94%(50 persons) mentors and 81%(17 people) department directors, eight-year program medical graduates were regarded as lower-skilled clinicians compared with doctoral graduates with professional degree . Eight-year program medical graduates were also considered with worse research capabilities than doctoral graduates with profes-sional degree by themselves (76%, n=56), their tutors (91%, n=48), and their department directors (86%, n=18). Conclusions The survey shows that to some extent, the current eight-year clinical doctoral graduates lack of career development potential, reflecting the current training mode cannot lead to the given training goals. We suggest that both clinical skills and research capabilities should be enhanced to make sure that eight-year program medical graduates deserve the title of academic physicianorphysician scientists.

Objective To investigate the clinical effect of S-1 combined with oxaliplatin (SOX regimen) in treatment of the patients with advanced pancreatic cancer. Methods A total of 106 patients with advanced pancreatic cancer in Qingdao Fuwai Hospital from April 2015 to June 2017 were randomly divided into the treatment group (53 cases) and the control group (53 cases) according to the random number table method. Patients in the control group were treated with S-1 combined with cisplatin treatment, and patients in the treatment group were treated with SOX regimen. The cell proportion of CD3+, CD4+, CD8+and CD4+/CD8+before treatment and after 2 cycles of treatment were detected by using flow cytometry of both groups. The clinical curative effects, immunity and adverse reactions of both groups were compared by usingχ2 test and t test. Kaplan-Meier method was used to make the survival analysis. Results After two cycles of treatment, there were 4 cases of complete remission (CR), 23 cases of partial remission (PR), 17 cases of stable disease (SD), 9 cases of progression disease (PD) in the treatment group, and 0 case of CR, 18 cases of PR, 20 cases of SD, 15 cases of PD in the control group. The rate of CR+PR in the treatment group was higher than that in the control group [50.94％(27/53) vs. 33.96％(18/53)], and there was a statistical difference (χ2=5.936, P<0.05). There was a significant difference in the cell proportion of CD4+and ratio of CD4+/CD8+between the two groups before and after treatment [the treatment group: (27.31±2.48)％ vs. (37.05±2.53)％, χ2= 6.491,P< 0.01; 0.91 ±0.23 vs. 1.53 ±0.50, χ2 = 5.913, P< 0.01; the control group: (27.43 ±2.47)％ vs. (30.32 ± 2.41)％,χ2= 11.214, P<0.01; 0.90±0.22 vs. 1.22±0.34,χ2=7.992, P<0.01]. After 2 cycles of treatment, the cell proportion of CD4+and ratio of CD4+/CD8+of the treatment group were higher than those of the control group, and there were statistical differences (χ2=5.309, P<0.01;χ2= 7.112, P< 0.01). The incidence rate of side effects had no significant difference in both groups after two cycles of treatment [22.64％ (12/53) vs. 18.87％ (10/53), χ2= 1.924, P> 0.05]. The progression-free survival time in the treatment group was longer than that in the control group (P<0.05). Conclusions SOX regimen has a favorable effect on the patients with advanced pancreatic cancer. It can help to improve the immunity and prolong the survival time of the patients.

OBJECTIVETo investigate the cell proliferation and genome stability in workers with occupational exposure to diesel exhaust (DE).

METHODSIn 2012, 117 DE-exposed workers and 106 control workers were recruited by cluster sampling in this study. The demographic data were obtained by questionnaire survey. The airborne fine particle and enriched polycyclic aromatic hydrocarbons (PAHs) at different workplaces were collected and analyzed. The concentrations of main PAHs monohydroxy metabolites in the urine were determined by high performance liquid chromatography-mass spectrometry (LC-MS), which could reflect the internal exposure level of DE. The cell proliferation capacity and genome stability in the periphery lymphocytes of workers were evaluated by cytokinesis-block micronucleus (CBMN) cytome assay.

RESULTSThe concentrations (median (P5-P95)) of total PAHs monohydroxy metabolites in the urine of exposed group and control group were 12.96 (4.73-28.10), 4.76 (0.90-15.00) µg/L, respectively, and the exposed group was higher than that of controls (Z = -8.77, P < 0.001). The nuclear division index (NDI) of exposed group and control group was 1.68±0.13, 1.85±0.16, respectively, and the NDI of exposed group showed significantly decreased (t = 8.86, P < 0.001), while the genome instability index calculated by micronucleus, nuclear bridges and nuclear buds, of exposed group and control group was 13.27±6.26, 4.83±3.38, respectively, and the exposed group had statistically significant increase (Z = -10.08, P < 0.001). The tertiles of total PAHs monohydroxy metabolites in the urine were categorized into low, medium and high groups (<5.96, 5.96-12.46, >12.46 µg/L). With the NDI decreased, 1.81±0.17, 1.79±0.17, 1.68±0.14 (F = 13.14, P < 0.001), genome instability index began to increase 5.80±4.15, 9.97±7.14, 11.99±6.61 (/1 000), respectively (χ(2) = 36.74, P < 0.001). With the increase of total PAHs monohydroxy metabolites level in corresponding groups. In addition, the NDI was negatively correlated with the frequencies of micronucleus, nuclear bridges, nuclear buds and genome instability index, respectively, and the difference was statistically significant (P < 0.001).

CONCLUSIONSDE exposure lead to inhibition of cell proliferation capacity and increase genome instability in the peripheral lymphocytes of occupational-exposed population, providing important clues and evidence for early biomarkers monitoring.

Biomarkers ; Cell Proliferation ; Chromatography, Liquid ; Genomic Instability ; Humans ; Lymphocytes ; Micronucleus Tests ; Occupational Exposure ; Particulate Matter ; Polycyclic Aromatic Hydrocarbons ; Surveys and Questionnaires ; Vehicle Emissions ; Workplace

Objective: To investigate the association between etheno-DNA adduct and the promoter of DNA methylation levels of cyclin dependent kinase inhibitor 2A (P16), Ras association domain family 1 (RASSF1A) and O-6-methylguanine-DNA methyltransferase (MGMT) in workers with occupational exposure to diesel engine exhaust (DEE). Methods: We recruited 124 diesel engine testing workers as DEE exposure group and 112 water pump operator in the same area as control group in Henan province in 2012 using cluster sampling. The demographic data were obtained by questionnaire survey; urine after work and venous blood samples were collected from each subject. The urinary etheno-DNA adducts were detected using UPLC-MS/MS, including 1,N6-etheno-2'-deoxyadenosine (εdA) and 3,N4-etheno-2'-deoxycytidine(εdC). The DNA methylation levels of P16, RASSF1A, and MGMT were evaluated using bisulfite-pyrosequencing assay. The percentage of methylation was expressed as the 5-methylcytosine (5mC) over the sum of cytosines (%5mC). Spearman correlation and multiple linear regression were applied to analyze the association between etheno-DNA adducts and DNA methylation of P16, RASSF1A, and MGMT. Results: The median (P₂₅-P₇₅) of urinary εdA level was 230.00 (98.04-470.91) pmol/g creatinine in DEE exposure group, and 102.10 (49.95-194.48) creatinine in control group. The level of εdA was higher in DEE exposure group than control group (P<0.001). DNA methylation levels of P16, RASSF1A and MGMT were 2.04±0.41, 2.19 (1.94-2.51), 2.22 (1.94-2.46)%5mC in exposure group, and 2.19±0.40, 2.41 (2.11-2.67), 2.44 (2.15-2.91)%5mC in control group. DNA methylation levels were lower in exposure group (P values were 0.005, 0.002 and 0.001, respectively). Spearman correlation analysis showed that DNA methylation levels of P16, RASSF1A, and MGMT were negative associated with urinary εdA level (r values were -0.155, -0.137, and -0.198, respectively, P<0.05). No significant correlation was observed between the εdC level and any measured DNA methylation levels (P>0.05) . Multiple linear regression confirmed the negative correlation between εdA and DNA methylation levels of P16, RASSF1A, and MGMT in non-smoking group (β (95%CI) was -0.068 (-0.132--0.003), -0.082 (-0.159--0.004) and -0.048 (-0.090--0.007), P values were 0.039, 0.039 and 0.024, respectively). Moreover, εdC was negative associated with DNA methylation level of MGMT in non-smoking group (β (95%CI) was -0.094 (-0.179--0.008), P=0.032). Conclusion DEE exposure could induce the increased of εdA and decreased of DNA methylation levels of P16, RASSF1A and MGMT.

Objective:To evaluate the diagnostic efficacies of computed tomography (CT), clinical manifestations and 2019 novel coronavirus (2019-nCoV) nucleic acid test results for the screening and diagnosis of novel coronavirus pneumonia.Methods:The clinical data of suspected cases with corona virus disease 2019 (COVID-19) visited fever clinic or stayed in emergency room of the First Affiliated Hospital of Nanchang University from January 23 to February 9, 2020 were collected. Totally 274 cases who met the inclusion criteria were enrolled. Four screening schemes including chest CT screening, scoring, chest CT and scoring series screening, chest CT and scoring parallel screening were designed. The statistical analysis was performed by using chi-square test. The sensitivities, specificities and the areas under the receiver operator characteristic curve of the four screening schemes were calculated, and the diagnostic efficacies were evaluated.Results:Among the 274 cases, 93.80%(257/274) presented with typical clinical symptoms, 47.81%(131/274) had epidemiological history, and the blood routine examination results of 45.26%(124/274) cases met the positive criteria of the scoring , and chest CT results of 43.07%(118/274) cases met the positive criteria of chest CT screening. The 2019-nCoV nucleic acid test positive rate of cases with epidemiological history was 30.53%(40/131), which was higher than that of cases without epidemiological history (9.09%(13/143)), and that of cases with typical imaging performance on chest CT was 40.68%(48/118), which was higher than cases without typical imaging performance (3.21%(5/156)). The differences between the above groups were both statistically significant ( χ2=20.150、60.468, respectively, both P<0.01). The positive rates of viral nucleic acid detection in cases with positive findings of chest CT screening, scoring, series screening, and parallel screening were 40.68%(48/118), 23.74%(47/198), 44.68%(42/94) and 23.87%(53/222), respectively, while those in cases with negative findings of the four screening schemes were 3.20%(5/156), 7.89%(6/76), 6.11%(11/180) and 0(0/52), respectively. The positive rates of the four screening schemes were all significantly different from that of viral nucleic acid detection ( κ=0.402, 0.100, 0.431, 0.106, all P<0.01). The chest CT screening method had a sensitivity of 90.57%and a specificity of 68.33%, and an area under the operating characteristic curve of the subject was 0.794, which had higher diagnostic value than those of the other three screening schemes. Conclusions:For the screening and diagnosis of COVID-19 cases, epidemiological history, positive 2019-nCoV nucleic acid test with ≥2 typical clinical manifestations have highly diagnostic value. On the basis of the preliminary screening of chest CT examination, flexible analysis of the diagnostic results could improve the diagnostic value of each detection method.

Objective:To investigate the early clinical effect of fascia lata autograft bridging combined with the long head of biceps tendon transposition for treatment of irreparable massive rotator cuff tear.Methods:All of 31 cases of massive irreparable rotator cuff tear treated in our hospital from March 2016 to March 2020 were analyzed retrospectively. Among them, 17 cases (10 males, 7 females) were repaired with fascia lata autograft bridging under arthroscopy (patch group), the average age was 61.47±6.63 (ranging from 51 to 72) and 14 cases (4 males, 10 females) were repaired with fascia lata autograft bridging combined with the long head of biceps tendon transposition (combined group), the average age was 62.57±6.11 (ranging from 53 to 71). The operation time, intraoperative blood loss, postoperative complications, visual analogue scale (VAS) of pain before operation, at 1 week and 12 months after operation, Constant-Murley score of shoulder joint and American Association of shoulder and elbow Surgeons (ASES) score before operation, at 6 months and 12 months after operation were compared between the two groups. The outcome of rotator cuff healing was evaluated by MRI 1 year after operation.Results:All patients were followed up for 12-27 months (mean 18.33 ±6.8 months). There was no perioperative complication, and there was no significant difference in operation time between the two groups ( P>0.05) . The VAS score in the patch group was significantly higher than the combined group 1 week after operation ( t=2.09, P=0.048) , and there was no significant difference in VAS score 12 months after operation between the two groups. Constant-Murley score and ASES score in the combined group were significantly higher than the patch group at 6 months after operation ( t=5.23, P<0.001; t=4.45, P<0.001) , and there was no significant difference in Constant score and ASES score between the two groups at 12 months after operation. Constant score and ASES score in the two groups were significantly higher than those before operation. One year after operation, the MRI of the affected shoulder showed that the incidence of autograft patch thinning (Sugaya grade III) was 52.94%, the autograft patch structure failure rate (Sugaya grade IV and V) was 17.65% in the patch group, the autograft patch thinning rate (Sugaya grade III) was 35.71%, and the structural failure rate (Sugaya grade IV and V) was 7.14% in the combined group. The difference was statistically significant (χ 2=7.12, P=0.028) . Conclusion:Fascia lata autograft patch bridging combined with long head of biceps tendon transposition technique for treatment of irreparable massive rotator cuff tear has less pain 1 week after operation and better recovery of shoulder function half a year after operation. MRI showed better patch healing 1 year after operation.

Objective: To summarize the incidence, clinical manifestations, diagnosis and treatment of basal cell nevus syndrome and to provide reference for clinical diagnosis and treatment. Methods : Retrospective analysis of 4 cases of basal cell nevus syndrome admitted to the General Hospital of PLA during January 2017 to January 2018 and recent cases reported in the literature. Results: In this study, 1 males and 3 females were included. The patients included a mother and her child. All 4 cases were surgically resected. Pathological reports included all keratocysts of the jaws. There has been no recurrence since follow-up. Through literature summarization and analysis, the clinical manifestations of this syndrome were found to be diverse. Typical clinical manifestations include multiple keratocysts of the jaws, multiple blepharospasms or cancers, deformities of the spine or ribs, increased brachial distance, eye diseases or special face intracranial calcification. Conclusion Basal cell nevus syndrome is an autosomal dominant genetic disorder. The clinical manifestations are diverse and the diagnosis is often overlooked. The incidence of cysts in the jaws is one of the important clinical manifestations of this syndrome. Early diagnosis and proper treatment improve patient survival and quality of life.