BACKGROUND:Adult central nervous system lacks the ability to regenerate,so it is of great significance to find a new source of neural stem cells.OBJECTIVE:To investigate the differences between basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)in inducing bone marrow mesenchymal stem cells(MSCs)to differentiate into neurons in vitro.METHODS:MSCs were isolated from normal human bone marrow using density gradient centrifugation and cell attachment method.MSCs were plated in 96-well culture plates at a density of 0.25×108/L and cultured with 200 μL DMEM/F12 for 0,1,2,3,4,5,6,and 7 days,with 20 μL MTT(5 mg/mL)in 5 wells at each time point.The supernatant was removed and 100 μL dimethyl sulphoxide was added to each well for 4 additional hours of incubation.In addition,some cells were exposed to bFGF and EGF.Growth curve was determined with MTT method.Telomerase activity were examined by TRAP(PCR)-ELISA.Additionally,the functional differences of the two cytokines were checked by RT-PCR.RESULTS AND CONCLUSION:RT-PCR revealed that nestin,glial fibrillary acidic protein(GFAP)and neurofilament subunit M (NF-M)mRNA were expressed in un-induced MSCs of passage 4.Nestin expression reduced at 7 days.The expression of micro-tubule-associated protein-2(MAP2)mRNA was not detected until the induction,and increased thereafter.The expression of MAP2 mRNA was greater in bFGF+EGF and bFGF alone groups compared with EGF alone group,and the expression of GFAP in EGF alone group was greater than other groups.Results showed that MSCs can be cultivated,proliferated and differentiated into neural stem cells in vitro.The differentiated neural stem cells have the activity of proliferation,but not have the ability of infinite proliferation as tumor cells.

Objective To investigate whether the extact of ginko biloba (EGb761) can protect the retinal ganglion cells (RGC) after acute high intraocular pressure induced retinal ischemia-reperfusion injury. Methods Thirty Sprague-Dawley rats were randomly divided into three groups. The negative control group, positive control group, treatment group were given normal saline 5 mL/kg, 1% erigeron breviscapus (vant) hand-mazz (EBHM) 5 mL/kg, 1% EGb761 5 mL/kg, respectively 2 hour after injury and every day after operation. Retinal ganglion cells were retrograde labeled by injection of 3% fluorogold (FG) into both side of superior colliculus 5 days before the death of the rats. Results The average survival rate of retinal ganglion cells were 61.46%, 72.73% and 76.20% in negative control group, positive control group and treatment group respectively. Conclusion EGb761 has neuroprotective effect on retinal ganglion cells in acute high intraocular pressure induced retinal ischemia-reperfusion injury.

OBJECTIVE:To evaluate the effect of special rectification on the rational use of antibiotics and relieving bacterial resistance. METHODS:It was divided into groups based on the before and after 3 years of special rectification. The data of related index and bacterial resistance of antibiotics in the clinical use in 2 groups were compared. RESULTS:After special rectification, the use rate of antibiotics in inpatients was decreased from 77%to 55%,use intensity was decreased from 86 DDDs/(100 person· d) to 39 DDDs/(100 person·d),the prophylactic use rate of antibiotics for typeⅠincision surgery was decreased from 98% to 27%,the antibiotics prescriptions of patients in outpatient and emergency departments were respectively decreased from 36% to 12% and 49% to 23%. The submission rate of microbiological testing specimens was increased from 20.2% to 38.8%,submission rate of sterile site specimens was increased from 29.8% to 37.6%. The detection rate of fungus was decreased from 14.4% to 11.2%. The detection rates of Escherichia coli and Klebsiella pneumoniae producing extended-spectrum β- lactamases (ESBLs) strains were respectively decreased from 57.3% to 51.3% and 43.2% to 36.1%. The total detection rate of top 5 multi-drug resis-tant bacteria was decreased from 48.4% to 29.3%,however,the detection rate of Acinetobacter baumannii was increased to 80.7%,and the resistance rates of Acinetobacter baumannii to imipenem and meropenem were respectively 66.9% and 69.1%. There was an increasing trend for Klebsiella pneumoniae to 1,2 and 3-generation cephalosporins,piperacillin amoxicillin/tazobac-tam,imipenem,meropenem. CONCLUSIONS:Special rectification of antibiotics has obvious effect on the rational use of antibiot-ics and relieving bacterial resistance in the clinic,and it improves the management of clinical use of antibiotics in hospital. Howev-er,bacterial resistance situation is still grim,it needs to establish a long-term management mechanism of clinical use of antibiotics, strengthen the monitoring pathologic examination and monitoring of bacterial resistance,and strictly perform hand hygiene and dis-infection and isolation system.

Objective To explore the impact of blockade of SDF-1/CXCR4 signaling pathway by T140 on degradation of typeⅡ collagen in human articular cartilage and to define the mechanism of action of T140. Methods 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from osteoarthritis patients undergoing total knee replacement ( OA cartilage group) and 144 pieces of cartilage (Mankin score of 0 or 1) were obtained from patients receiving traumatic amputation (normal cartilage group). Each group was treated with SDF-1 of 100 ng/mL, then divided into three subgroups A, B, and C. The cartilage tissue in each group was cultured in the nutrient solution containing of T140, MAB310, or SDF-1 (1 000 nmol/L) for 48 or 96 hours. RT-PCR was used to detect expression of typeⅡcollagen mRNA in the cartilage tissue. Results Level of type Ⅱcollagen mRNA was markedly higher in subgroup A than in subgroup B and subgroup C at the same group and the same time (P <0.05). The expression level of type Ⅱcollagen mRNA at the same time and in the same subgroup of OA cartilage group were lower than that in normal cartilage group (P < 0.05). Conclusions SDF-1 induces degradation of typeⅡcollagen in human articular cartilage thruogh the SDF-1/CXCR4 signaling pathway. T140 can block the SDF-1/CXCR4 signaling pathway and reduce the degradation of type II collagen.

ObjectiveTo study the toxin genes and pandemic group distribution as well as the genetic correlation between the major serotypes( O3:K6,O1:Kut,O4:K8 ) of Vibrio parahaemolyticus (VP) isolated from the outbreaks of Guangdong province.MethodsThe tdh and trh genes,GS-PCR and orf8 gene were detected on the 62 isolates sourced from patient and seafood occurred in the 23 outbreaks during 2008-2010.44 isolates of which were analyzed on PFGE digested by Not Ⅰ enzyme.ResultsToxin genes distributions suggested that 96.8% (60/62)isolates were tdh+,trh-.Three tdh+ isolates sourced from seafood were found.Pandemic group distribution suggested that 97.2% (35/36) O3:K6,5.88% (1/17) O4:K8,66.7% (8/9)O1:Kut serotype was GS-PCR+ and/or orf8+,respectively.PFGE analysis suggested that 44 isolates were separated into 3 clusters,of which the similarity of PFGE profile was 80.5% in the pandemic group cluster constructed by 28 isolates,the similarity between pandemic group and non-pandemic group was 59.5%.Pandemic group of O3:K6,O1:Kut as well as 04:K8 isolated on some outbreaks were processing the same PFGE profiles.ConclusionThe characteristic of toxin genes of major serotypes VP isolated in the outbreaks of Guangdong province form 2008-2010 was tdh- present and trh- absent.Within the pandemic group,O3:K6 and O1:Kut were the major serotypes.In single outbreak,isolates belongs to pandemic group but with different serotype seem to be close correlations.

OBJECTIVE To observe the curative effect of external dacryocystorhinostomy(EDCR) and intranasal endoscopic dacryocystorhinostomy(IEDCR)for chronic dacryocystitis, and to evaluatethe postoperative satisfaction of the patients.METHODS According to randomized controlled design, 108 chronic dacryocystitis patients were divided into the intranasal endoscopic dacryocystorhinostomy group and control group, both groups underwent intranasal dacryocystorhinostomy and external dacryocystorhinostomy respectively.The changes of symptom and clinical sign, operation time, hospital day, cost of hospitalization were recorded.All patients were followed up for 1-5years, and the postoperative satisfaction of the patients were investigated by interval satisfaction scale.RESULTS ①There were no significant difference between the groups at the curative effect, operation time and hospital day, the hospitalization cost of the intranasal endoscopic dacryocystorhinostomy group was higher than that of the control group;②The recurrence rate of the intranasal endoscopic dacryocystorhinostomy group and control group were 12.82% and 15.56% respectively after 5 years, Log Rank test ?2=0.394, P=0.530;③ The postoperative satisfaction of the patients in both groups were(81.70?10.242)and(75.72?10.653) score respectively, which were similar to normal distribution, t=-2.974, P=0.004.CONCLUSION The effect of intranasal endoscopic dacryocystorhinostomy for chronic dacryocystitis is similar to external dacryocystorhinostomy.But intranasal endoscopic dacryocystorhinostomy is a more ideal method, because it is minimal invasive technique and doesn't affect the face cosmetology.

BACKGROUND:Human mesenchymal stem cells (hMSCs) become aging and even die after several passages. Some investigations have shown that telomere has a close correlation with life span of the cells. Whether the ectopic expression of human telomerase reverse transcriptase (hTERT) could induce the activity of the telomerase, maintain the length of telomere, and finally prolong the life cycle of MSCs without losing their multipotent differentiation capacity is still uncertain.OBJECTIVE: To observe the influence of the ectopic expression of hTERT on the telomerase activity and cell life cycles of hMSCs.DESIGN: Repetitive measurement trails.SETTING: Research Center of Stem Cell, Zhengzhou University Medical College.MATERIALS: The experiment was conducted in the Research Center of Stem Cell, Zhengzhou University Medical College from October 2003 to December 2005. hMSCs were obtained from 20 healthy donators from the Department of Pediatric Surgery and Outpatient, the Third and First Affiliated Hospitals of Zhengzhou University. Enhanced green fluorescent protein plasmid (pEGFP-C1) and pEGFP-hTERT were provided by Dr. Chantal Autexier of Canada. DH5α strain provided by Dr. Hou Wei-hong, the Key Molecular Medical Laboratory of Zhengzhou University Medical College.METHODS.: Under sterile condition, 2 mL bone marrow of sternum of healthy donors were harvested, and prepared after centrifugalization,dilution and passage.① Transfection of pEGFP-hTERT into hMSCs and the screening and amplification of resistance cloning:The 5th passage cells were seeded in a 24-well plate,and transfected by pEGFP-hTERT with lipofectamine method.The cells were divided into four groups including untransfected group,lipofectamine group,pEGFP-C1 group and pEGFP-hTERT group. Resistance cloning screen and amplification was performed by G418. ②hTERT mRNA expression and detection of telomerase activity:RT-PCR and PCR-ELISA were used to detect the hTERT mRNA expressions of the fifth passage hMSCs transfected with pEGFP-hTERT, and pEGFP-C1, the untransfected tenth passage hMSCs and K562 cells (positive control), and the telomerase activity of the fifth and thirtieth passage hMSCs transfected with pEGFP-hTERT,the fifth pEGFP-C1-transfected cells and the tenth passage untransfected cells. ③Karyotype analysis of hTERT-transfected MSCs: Chromosome analysis was performed by conventional Giemsa staining.④Inducement of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification:The transfected MSCs were cultured in a medium containing epidermal growth factor and basic fibroblast growth factor, which could induce the cells differentiate into neuron-like cells. The culture solution was changed every 3 days, and the changes in cell growth condition and morphologic characteristics were observed under an inverted microscope. The microtubule associate protein (MAP2) and neurofliament subunit M (NF-M) were identified by RT-PCR.MAIN OUTCOME MEASURES:①hMSCs transfection with different kathion liposomes and the screening and amplification of resistance cloning; ②hTERT mRNA expressions of each group and detection of telomerase activity; ③Karyotype analysis of pEGFP-hTERT-transfected MSCs; ④ Induction of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification.RESULTS: ①With the decrease of G418 concentration, the cells in the untransfected and lipofectamine groups died, and stably EGFP expressed MSCs were obtained; after G418 screening, there was a cell clone undergone 35 passages and continued to proliferate, whose appearance and growth characteristics were similar to the untransfected MSCs observed under inverted microscope. ②The fifth passage pEGFP-C1-transfected hMSCs and tenth passage untransfected hMSCs remained telomerase-negative, but the K562 and fifth passage hTERT-transfected cells showed positive telomerase activity. ③The telomerase activity of the fifth and thirtieth passage hTERT-transfected cells was positive. ④The hTERT-MSCs at passage 10, 20 and 30 had 23 pairs of chromosomes, and two X chromosomes. So they were still normal diploid with normal chromosome appearance and number. ⑤Many hTERT-transfected MSCs had the typical appearance of neuron-like cells. RT-PCR analysis showed that th expressions of MAP2 and NF-M were increased.CONCLUSION:Ectopic expression of the hTERT gene is found in hMSCs,and can induce the telomerase activity of hMSCs.The ectopic expression of the hTERT gene in hMSCs could extend the life spans of cells and maintain their multipotent differentiation capacity.

To study the value of high-fluorescent cell for the screening of malignant hydrothorax or ascites. The clinical diagnosis results and hydrothorax or ascites specimens of 400 outpatients registered from January to December in 2014 were collected, and high-fluorescent cells were counted with Sysmex XE-5000 blood cell analyzer, and then stained for microscopy. Receiver operating characteristic (ROC) curve was used to explore the sensitivity, speci-ficity, positive likelihood ratio and negative likelihood ratio of high-fluorescent cell for screening malignant hydrothorax or ascites. The numbers of high-fluorescent cells of the cancer patients were significantly higher than those of the non-cancer patients, with Z equal to -7.372 and P less than 0.05. The values of the area under curve (AUC), sensitivity, specificity, positive likelihood ratio and negative likelihood ratio were 0.801, 70.67%, 84.31%, 4.50 and 0.35 respective-ly. Detection of high-fluorescent may be used to screen malignant hydrothorax or ascites, and further exami-nation and follow-up have to be performed in case the number of high-fluorescent cells in the hydrothorax or ascites is not less than 68.

To retrospectively analyze the clinical data of 42 patients at our department with bulbar myasthenia gravis.They underwent video-assisted thoracoscopic extended thymectomy (VATET) from May 2003 to August 2010.Another 42 patients treated with medications alone were selected as controls.Compared with the control group (4.32 ± 0.23),postoperative dysphagia scores improved significantly in patients undergoing VATET(9.21 ±0.41).And so did the dysphagia assessment classification (P ＜0.05).Thus VATET could effectively and definitely relieve dysphagia in patients with bulbar myasthenia gravis.

Objective Objective To study the effect of Mycobacterium tuberculosis (Mtb) on IFN-? signaling pathway in macrophages. Methods Humans and mice macrophages were isolated and infected by Mtb, condition medium were collected after a few days culture; Macrophage surface expression of Fc?RI (human) or MHC II (mouse) was measured by flow cytometry to explore the effect of condition medium on macrophage response to IFN-?. Results Surface expression of Fc?RI in human macrophages cultured in condition medium was significantly decreased compared to those without condition medium (p