Objective To investigate the protective effect and the mechanisms of 17β-estradiol on ketamine-induced apoptosis on primary cultured rat cortical neurons. Methods Cortical neurons were primarily cultured for seven days,then divided into four groups :control group ( treated with equal valume of DMSO ),estradiol-treated group ( treated with 0.1 μmol·L-1 17β-estradiol),ketamine-treated group(treated with 100 μmol·L-1 ketamine),ketamine plus 17β-estradiol-treated group( treated with 0. 1 μmol·L-1 17β-estradiol+100μmol·L-1 ketamine). The neurons were treated for 24 hours. The neuron viability was determined by MTT. Neuroapoptosis was measured by nuclear morphometry after Hoechest 33258 dying. Western blotting was performed to detect the expression levels of cleaved-caspase-3 and Bcl-2protein. Results The neuron viability in the ketamine group was(54. 02±7. 78)%,significantly decreased from the control group,whereas ketamine plus 17β-estradiol increased the cell viability to(88. 09±6. 54)%,significantly higher than the ketamine group. The neuroapoptosis rate in the ketamine group was(49. 50±4. 34)%,significantly increased from the control group,while that in the drug combination group was(15. 74 ± 3. 40)%,significantly lower compared with the ketamine group. Meanwhile,the cleaved-caspase-3 expression increased,and Bcl-2 expression decreased remarkably after ketamine treatment,while which was reversed in the drug combination group. Conclusion 17β-estradiol can protect against ketamine-induced injury by inhibiting neuron apoptosis.

Objective To investigate the mechanisms of the protective effects of dexmedetomidine against the propofol-induced neuroapoptosis in primary cultured cortical neurons.Methods The neurons were cultured 7days and then divided into four groups: vehicle-control group (treated with equal volume of intralipid), propofoltreated group (treated with 500 μmol/L propofol), propofol plus dexmedetomidine treated group (treated with 500 μmol/L propofol and 0.1 μmol/L dexmedetomidine), and LY294002 pretreated group (treated with 500 μmol/L propofol ,0.1 μ mol/L dexmedetomidine and 10 μmol/L LY294002).12 hours after different treatments, neuron viability was measured by MTT assay,neuroapoptosis was detected by Hoechst33258 staining, and the levels of pAkt and Bcl-2 protein were detected by Western blot.Results Compared with the vehicle-reduced group,propofol reduced neuron viability greatly((53.4±4.2)％ vs (99.9±6.3)％;P＜0.01), but increased neuroapoptosis greatly((44.6±4.3)％ vs (5.8±0.4)％;P＜0.01).The levels of pAkt((0.41±0.03) vs (0.86±0.07))and Bcl-2 ((0.15±0.02) vs (0.72±0.03)) were decreased greatly (both P＜0.01).Compared with propofol treatment group, the neuron viability of propofol plus dexmedetomidine group were increased greatly((86.4±5.3) ％ , P＜0.01) ,the neu roapoptosis was decreased greatly ((23.1 ± 3.5) ％, P＜ 0.01), and the levels of pA kt (0.8 ± 0.03) and Bc1-2 (0.52 ±0.05) were increased greatly (both P＜0.01).Compared with propofol plus dexmedetomidine treated group,LY294002 inhibited the protective effects of dexmedetomidine, decreased neuron viability greatly ((64.3±5.1) ％ ,P＜0.01), increased the number of apoptotic neurons((38.8±4.9) ％, P＜0.01), and reduced the levels of pAkt (0.52±0.04) and Bcl-2(0.31±0.02) significantly (P＜0.01).Conclusion Dexmedetomidine exerts the neuroprotective effects against propofol-induced neuroapoptosis by activating the PI3K-Akt-Bcl-2 signalling pathway.

Objective To investigate the effect of propofol exposure on neuroapoptosis in pri-mary cultured cortical neurons and its mechanisms.Methods Cortical neurons were primarily cultured for seven days,then divided into two groups:control group (treated with equal volume of 20% in-tralipid),propofol-treated group (treated with 500 μmol/L propofol).The neurons were treated for 12 h.The neuron viability was determined by MTT.Neuroapoptosis was identified by Hoechest 33 258 dying.Mitochondrial membrane potential was measured by the fluorescent dye rhodamine 123 (Rh123).Western blot was performed to detect the level of cyt-c and cleaved-caspase-3.Results Neu-rons survival rate (54.4%±6.4%)in the propofol group was significantly lower than that of control group (99.8% ± 4.1%) (P < 0.05 ), the rate of neuronal apoptosis (46.5% ± 5.3%) was significantly higher than that of control group (7.2%±0.9%)(P <0.05),mitochondrial membrane potential (59.6%±4.3%)was significantly lower than that of the control group (99.9% ± 5.7%) (P <0.05 ),cyt-C protein level (0.38 ± 0.03 )was significantly higher than that of control group (0.1 5±0.02)(P < 0.05 ),level of cleaved-caspase-3 protein level (0.46 ± 0.04)was significantly higher than that of control group (0.13±0.02)(P <0.05).Conclusion Propofol induces neuroapo-tosis in primary cultured cortical neurons,which is associated with the decreased level of MPP and the increase levels of cyt-c and cleaved-caspase-3.

Objectve To investigate the application value of ultraifltration with millipore ifltration method test of old biological samples DNA. Methods 23 old blood samples separately were cut blood piece equally suitable size of three groups, labeled A, B and C group. DNA was extracted by magnetic bead methed and get DNA template with 80μL, 80μL, 20μL elution solution, respectively, then the DNA template in group A and C take right amount directly ampliifed, group B with milipore ultraifltration ifltration condensed amplication. PCR products were detected by ABI3130xl genetic analyzer and analyzed by GeneMapperID V3.2 software. The date were analyzed by using SPSS software. Results No samples were ampliifed for all STR loci in group A. There were 18 cases in group B samples amplified all STR loci and 11 samples in group C. Conclusion Application of millipore ultrafiltration method can signiifcantly improve the old biological samples DNA classiifcation success rate.

OBJECTIVE To understand the situation of application of antibacterials in perioperative period in Department of Orthopedics,in order to find evidence for strengthening the rational use and standard management of antibacterial.METHODS A total of 308 cases with orthopedic clean-operations were randomly selected for the analysis of rationality of application of antibiotic prophylaxis in perioperative period.RESULTS All of the 308 patients were given antibiotics in perioperative period.And 18 kinds of drugs from 7 categories were used,the average kinds used were(2.07?0.75).The patients started antibiotic administration within 0.5-2 hours before operation accounted for 84.74%.The average time of antibiotic using was(6.07?2.64) days.72.40% patients in medication period used more than one kind drugs.CONCLUSIONS There are some problems in application of antibacterials in orthopedic clean-operations during perioperative period,such as too long-term use of antibiotics,frequent changes and so on.It is necessary to strengthen the management and improve the rationality of application of antibacterials.

Objective To investigate the protective effects and the mechanisms of 17β-estradiol on the propofol-induced neuroapoptosis in primary cultured rat cortical neurons.Methods The neurons were cultured for 7 days and then divided into three groups: vehicle-control group ( treated with equal volume of 20% intralipid ) , propofol-treated group ( treated with 500μmol/L propofol) , and propofol plus 17β-estradiol treated group ( treated with 500μmol/L propofol and 0.1 μmol/L 17β-estradiol).12 hours after the treatment, neuroapoptosis was detected by Hoechst 33258 staining and TUNEL assay, and the levels of Bcl-2, Bax and cleaved caspase-3 proteins were detected by Western blot.Results Compared with the vehicle-control group, the neuroapoptosis increased greatly ( P<0.01 ) , Bcl-2 level reduced ( P <0.01), Bax and cleaved caspase-3 levels increased greatly (P<0.01), and Bcl-2/Bax ratio reduced significantly (P<0.01).Compared with the propofol-treatment group, the neuroapoptosis decreased greatly ( P <0.01), Bcl-2 level increased ( P<0.01 ) , Bax and cleaved caspase-3 levels reduced greatly ( P <0.01 ) , and Bcl-2/Bax ratio increased greatly ( P <0.01 ) . Conclusions 17β-estradiol can protect cortical neurons against propofol-induced cortical neuroapoptosis by regulating the expression of Bcl-2 and Bax.

Objective To investigate the prevalence of toxoplasma gondii (Tox),rubella virus (RV),cytomegalovirus (CMV) and herpes simplex virus (HSV) infections (TORCH infections) among childbearing-age population in Henan province.Methods Enzyme-linked immunosorbent assay (ELISA) was applied to detect plasma TORCH IgM and IgG among 3084 childbearing-age men and women from theFirst Affiliated Hospital of Zhengzhou University during July and September,2011.The positive rates of anti-TORCH antibodies were compared among the various age and gender groups by x2 test.Results The total positive rate of anti-TORCH IgM was 5.5％ (170/3084),in which the positive rate of anti-RV IgM was the highest (2.9％),followed by anti-HSV IgM (1.0％).Within positive rate of anti-TORCH IgG,anti-HSV IgG was the highest (90.4％),followed by anti-CMV IgG (89.7％),RV IgG (48.1％) and Tox IgG (0.7％).The positive rate of anti-TORCH IgM was the lowest in individuals aged ＞ 30-40 year old.With the age increasing,the positive rates of anti-Tox IgG,anti-CMV IgG and anti-HSV IgG increased,but the positive rate of anti-RV IgG decreased.Women had higher positive rates of anti-CMV IgG and antiHSV IgG than men (x2 =83.470 and 7.026,P ＜ 0.O1).Conclusions Current infection of TORCH exists in childbearing-age population of Henan province,and the positive rate of anti-RV IgG is low.It is recommended to screen for TORCH infection in childbearing-age men and women.

Obje:ctive To establish an optimized method to isolate, culture and identify human umbilical cord mesenchymal stem cells (hUCMSCs) in vitro and induce their osteogenic and adipogenic differentiation. Methods The hUCMSCs were isolated from human umbilical cord by digestion with collagenase. After serial subcultivation in vitro, the stem cells were passaged. Morphologic appearance of hUCMSCs was observed under an optical microscope and atomic force microscope. The proliferation rate was measured by MTT assay. Cell cycle and surface antigens were measured by flow cytometry. The osteogenic and adipogenic differentiation was tested and evaluated by specific staining methods. Results The isolation of hUCMSCs by digestion with collagenase was efficient. After seeded for 24 hours, the adherent cells showed spindle shape and fibroblast cell-like shape and the size of hUCMSCs was homogeneous. The similar growth curves of passage 3 and 7 exhibited a great potential for proliferation. Flow cytometry analysis revealed that CD29, CD44 and CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, CD45 and HLA-DR. After culture in inducing medium, the cells were successfully induced into osteogenic and adipogenic lineages. These cells were highly positive for alkaline phosphate staining and also showed mineralization presented with von kossa staining after 4 weeks' culture induction of osteogenic differentiation. Furthermore, liquid vacuoles were detected by oil red O staining after 3 weeks' culture induction of adipogenic differentiation. Conclusion An in vitro method for isolation and purification of hUCMSCs from human umbilical cord has been established. The cultured cells were composed of only undifferentiated cells and their biological properties were stable. The hUCMSCs are expected to be a new type of stem cells of tissue engineering.

Objective To investigate the occurrence of postoperative healthcare-associated infection(HAI)in pa-tients with hepatobiliary malignant tumor,explore the related risk factors,so as to provide the basis for taking ef-fective prevention and control measures.Methods The occurrence of postoperative HAI in patients with hepatobili-ary malignant tumor in a hospital from January 2012 to December 2014 were retrospectively analyzed,risk factors for postoperative HAI were analyzed through reviewing and collecting patients’medical data.Results A total of 302 patients were investigated,42 (13.91 %)developed postoperative HAI,no multiple site infection occurred,the main infection site was deep surgical site (n=10,23.81 %),followed by lower respiratory tract (n=9,21 .43%) and digestive system (n=7,16.67%).Of 42 infection cases,38(90.48%)were sent specimens for pathogenic cul-ture,36 pathogenic strains were isolated,31 (86.11 %)of which were gram-negative bacteria,and 5 (13.89%) were gram-positive bacteria.Multivariate logistic analysis showed that operation duration≥2 hours (OR =1 .48), overweight (or obesity)(OR=1 .40),and preoperative radiotherapy (OR=2.98)were independent risk factors for postoperative HAI in patients with hepatobiliary malignant tumor (all P <0.05).Conclusion Incidence of postoper-ative HAI is high in patients with hepatobiliary malignant tumor,risk factors are long length of operation,over-weight (or obesity),and preoperative radiotherapy,effective prevention and control measures against risk factors should be taken.

Objective To investigate the effect of 17β-estradiol on ketamine-induced long-term cognitive deficits in neonatal rats.Methods 80 SD male rats aged 7 days were randomly divided into group C,V,E,K and K+E,and 16 per group.Group C was intraperitoneally injected with same volume of saline for three consecutive days,Group V was subcutaneously injected with same volume of sesame oil for three consecutive days,Group E was subcutaneously injected with 600 μg · kg-1 17β-estradiol for three consecutive days,group K was intraperitoneally injected with 75 mg · kg-1 ketamine for three consecutive days,group K+E was intraperitoneally injected with 75 mg · kg-1 ketamine in combination with 600 μg · kg-1 17β-estradiol injected subcutaneously for three consecutive days.At 2 months of age,learning and memory abilities were tested with the Mortis water maze.After Morris water naze test,ten rats from each group were decapitated and the prefrontal cortex (PFC) was isolated to detect acetylcholine esterase(AchE) activity with ELISA assay and to measure acetylcholine(Ach) level by hydroxylammonium chloride method.Results The escape latency ((40.26±2.36) s,(30.25±2.20) s,(21.55±2.42) s) and path length((1019.35±58.13) cm,(811.16±27.58) cm,(598.34±34.74) cm) of group K were more than those of group C on the third,fourth aud fifth training days (all P＜0.05),while escape latency ((29.46±2.20) s,(24.86± 2.14) s,(17.20±1.91) s) and path length((913.90±41.89) cm,(729.42±31.36) cm,(487.64±18.61)cm) of group K+E were significantly lower than those of group K(all P＜0.05).On test day 6,rats were subjected to a probe trial,ratio of time spent in the target quadrant ((24.5±2.7) ％) and the number of crossings over previous platform locations (1.9±0.5) in group K were fewer than those of group C (all P＜0.05),while ratio of time spent iu the target quadrant((42.3±3.0) ％) and the number of crossings over previous platform locations(3.5±0.5) of group K+E were more than those of group K (all P＜0.05).The AchE activity((0.69±0.04) U · mg pro-1) in rats PFC of group K was significantly higher than that of group C ((0.52±0.06) U · mg pro-1) (P＜0.05).The AchE activity of group K +E ((0.58±0.12)U · mg pro-1) was lower than that of group K(P＜0.05).The Ach level ((2.59±0.34)mg · g-1) in rats PFC of group K was significantly lower than that of group C ((4.35±0.56) mg · g-1) (P＜0.05).The Ach level of group K+E((3.88±0.61) mg · g-1) was higher than that of group K(P＜0.05).Conclusions These results indicate that ketamine impairs learning and memory abilities as rat matures,while 17β-estradiol treatment improves these impairments by inhibiting AchE activity and increasing Ach level.