Objective:To discuss a simple method for flow cytometric analysis of cell DNA stained with DAPI and Hoechst33342.Methods:HT 29 cells stained with DAPI,Hoechst33342 or PI were measured by BD FACSAria and the percentages of cells in G0/G1,S and G2/M phases with three staining me-thods,then the results were analyzed and compared.Before measurement we monitored the quality of DNA analysis of flow cytometer through UV beads QC experiment and analyzed the standard chicken erythrocyte nuclei(CEN) and calf thymocyte nuclei(CTN) stained with DAPI and Hoechst33342.Results:CV value of UV peak was 2.4 after QC experiments.There were 4 peaks on CEN histograms and the ratios of peak channel mean of G2/G1,G3/G1,and G4/G1 were about 2,3,and 4 respectively.Both CV values of the first peak were 2.4.There were 2 peaks on CTN histograms and the ratio of peak channel mean of G2/G1 was 1.97,and CV value of G0/G1 2.4.The complete cell cycle of HT29 cells stained with DAPI,Hoechst33342 or PI was showed entirely,CV values were 3.40,3.02 and 4.42,respectively,and the percentages of cells in G0/G1 were 60.86%,60.22% and 60.81%,respectively,in S,28.85%,29.70% and 29.82%,respectively,and in G2/M,10.29%,9.09% and 9.37%,respectively.The results by the three methods showed no difference.Conclusion:This method for measurement of cellular DNA content is a simple and efficient approach to determining cell cycle and can be the first choice when using flow cytometer with 355 nm UV.

OBJECTIVE To investigate and control the risk factors about hospital infection in patients with chronic severe hepatitis B. METHODS Totally 466 hospitalized patients with chronic severe hepatitis B in our hospital from Jan 2001 to Dec 2007 were studied retrospectively. RESULTS The total incidence of hospital infection was 20.1% for inpatients with chronic severe hepatitis B,the infection located as follows:56 patients were in lower respiratory tract (59.6%);17 patients in upper respiratory tract (18.1%);7 patients with bacteremia (7.4%);3 patients in urinary tract (3.2%);7 patients in gastrointestinal tract (7.4%);and 1 patients was in cutis tissue (1.1%). CONCLUSIONS The knowledge about the on infection risk factors liver disease must be strengthened by hospital medical staff of the use of antibiotics must be strict controlled in principle,operation must be aseptic,the concept of prevention standards must be enhanced,and the primary disease must be actively treated,all these can effectively reduce infection in hospital.

BACKGROUND:Neural stem cels have the potential to differentiate into neurons and glial cels to replace the injured brain cels, so as to achieve the purpose of repairing nerve injury.
OBJECTIVE:To observe the neuronal differentiation ability of cel subsets with stem cel characteristics in the adult rat meningeal tissues.
METHODS:Under anesthesia, the meningeal tissues were obtained from adult Sprague-Dawley rats to make cel suspension folowed by inoculation and subculture. Then, the Nestin immunofluorescence staining was performed. The third generation cels were culturedin vitro with complete culture medium containing trichostatin A. After 7 days of induction, western blot assay was used to detect the expression of NF-200 and BM88 proteins in neural cels.
RESULTS AND CONCLUSION: At 24 hours of culture, some spherical cels were suspended and some cels adherent. In addition, some spherical cels scattered gradualy formed the clone spheres, and the growth rate decreased with the increasing volume. The positive expression of Nestin was detected by immunocytochemistry staining, and the cel nucleus was stained blue by Hoechst staining. BM88 and NF-200 proteins were al expressed at 7 days of neural induction. These findings indicate that the cel subsets with stem cel characteristics in the adult rat meningeal tissues can differentiate into neurons after in vitro induction.

Objective To observe the clinical curative effect of patients with ovulation obstructive infertility by combined treatmentofstimulating ovulation particles and clmiphene citrateclinical. Methods 171 patients were randomly divided into Traditional Chinese medicine (TCM) group (42 cases), clmiphene citrateclinical (CC) group (53 cases) andcombined treatment of TCM and CC group (76 cases). Ultrasound examinations were used to monitor the development of endometrium, follicular development and value of Plusatility Index (PI). The levels of E2,LH and P were measured before and after medication. And pregnancy of patients was followed up. Results Ovulation rates of combined treatment of TCM and CC group and the CC group were both higher than TCM group (P<0.05). Pregnancy rate of combined treatment of TCM and CC group was higher than TCM group and CC group (P<0.05). Endometrial thickness and type A lining rate of combined treatment of TCM and CC groupandTCM group were both higher than CC group (P < 0.05). The values of PI of combined treatment of TCM and CC groupand TCMgroup were both lower than CC group on HCG day after treatment (P<0.05). E2 values ofpost-treatment of three groups were all significantly higher than that before treatment.Difference in values of E2 and LH between pretherapy and post-treatment of combined treatment of TCM and CCwere obviously higher than that of TCM group (P<0.05).Difference in values of P between pretherapy and post-treatment of tcombined treatment of TCM and CC and TCM group were higher than CC group on the seventh day after ovulation (P < 0.05). Conclusions Combination of traditional Chinese medicine and CCcouldnot only promote follicle growth , but also improve anti-estrogen role of CC , endometrial blood flow and endocrine regulation , enhance ovulation rate and decrease the rate of abortion. Ovulation obstructive infertility treatment was better than that of CC and TCM treatment.

Objective To observe the efficacy of Zhuang’s medicated thread moxibustion plus acupoint thread embedding in treating allergic rhinitis.Method By using random number table, 103 patients with allergic rhinitis were randomized into a treatment group (n=53) and a control group (n=50). The treatment group was intervened by Zhuang’s medicated thread moxibustion and acupoint thread embedding, while the control group was by acupoint thread embedding alone. The subjects were treated for 6 courses with a one-month follow-up. The therapeutic efficacies were compared between the two groups.Result The total effective rates were respectively 94.3% and 80.0% in the treatment group and control group.Conclusion Zhuang’s medicated thread moxibustion plus acupoint thread embedding has significant advantage in improving the symptoms of allergic rhinitis.

Objective To construct human surfactant protein B (SP-B) gene promoter luciferase reporter plasmids and detect their transcriptional activities in H441 cells.Methods (1)The fragment of SP-B promoter (-218/+ 435 bp) was acquired from human genome DNA by polymerase chain reaction (PCR) amplification and then was inserted into pGM-T vector by the T4 DNA ligase.The vector was transfected into TOP10 E.coli.The positive clone was identified by DNA sequencing.The identified target SP-B promoter sequence was cloned into pGL3-basic vector to construct the recombinant vector pGL3-basic-SP-B-promoter and was identified by enzyme digestion and sequencing; (2)The pGL3-basic-SP-B-promoter vector was converted into pGL4.17-SP-B-promoter vector through enzyme digestion.The identified recombinant vectors and control plasmid pRL-TK were transfected into H441 cells by lipofectamine 2000,and luciferase assays was performed using the dual-luciferase reporter assay system.Results The sequences of SP-B promoter in the recombinant luciferase reporter plasmids were consistent with the one published on Genebank.The firefly/renilla luciferase activity ratio of pGL3-basic/pGL4.17-SP-B-promoter vector (2.8 ± 1.1,66.5±3.8) was significantly higher than pGL3-Basic,pGL4.17 control vector (0.2 ±0.1,4.3 ±0.4) with statistical significance (t =4.182,27.419,P =0.000),respectively.The SP-B promoter activity of pGL4.17-SP-B-promoter vector was significantly higher than pGL3-basic-SP-B-promoter vector (t =27.712,P =0.000).Conclusions The pGL3-basic/pGL4.17-SP-B-promoter vectors are successfully constructed with SP-B promoter activity in H441 cells and pGL4.17-SP-B-promoter vector is the better choice for further study with higher luciferase activity.

Objective: To explore the clinical value of serum N-methyl-aspartate receptor (NMDAR) antibody level, brainstem auditory evoked potential (BAEP) and magnetic resonance imaging (MRI) in the differential diagnosis of viral encephalitis and anti-NMDAR encephalitis. Methods: The clinical data of 68 children patients with encephalitis were retrospectively analyzed. The patients diagnosed with viral encephalitis were included in V group (n=52), and the patients diagnosed with anti-NMDAR encephalitis were included in N group (n=16). The clinical characteristics, serum NMDAR antibody level, and BAEP and MRI findings were compared between the two groups. Results: The age, disease duration, abnormal behavior rate, sleep disorder rate and epileptic seizure rate in V group were significantly lower than those in N group [(6.62±1.20)Y/O vs.(8.46±1.85)Y/O, (3.53±0.71)d vs.(4.49±0.82)d, 30.77%(16/52)vs. 75.00%(12/16), 21.15%(11/52)vs. 62.50%(10/16), 26.92%(14/52)vs. 56.25%(9/16), t=4.681, t=4.560, χ²=9.882, χ²=7.958, χ²=4.701], while the abnormal rate of video EEG was significantly higher than that in N group [51.92(27/52)vs. 81.25%(13/16), χ²=4.345] (all P<0.05). There were no significant differences in gender, rates of prodromic infection symptoms, cognitive impairment, fever, headache, convulsion and incidence rate of meningeal irritation sign (P>0.05). The serum NMDAR antibody level in V group was significantly lower than that in N group [(3.40±0.69) ng/ml vs.(13.95±2.78) ng/ml t=25.319)] (P<0.05). There were no significant differences in the BAEP apparent involvement range and central auditory neurological damage between the two groups (P>0.05), but the peripheral auditory nerve damage and total BAEP abnormality rate in V group were significantly lower than those in N group [3.85%(4/104)vs. 21.88%(7/32), 6.73%(7/104)vs. 28.12%(9/32), 30.77%(16/52)vs. 62.50%(10/16), χ²=10.699, χ²=10.790, χ²=5.216] (all P<0.05). There were no significant differences in MRI signal intensity, lesion involvement range and total abnormal rate between the two groups (all P>0.05). Conclusions There were significant differences in serum NMDAR antibody level and BAEP test results among children patients with viral encephalitis or anti-NMDAR encephalitis, and they are helpful for early differential diagnosis.

Objective To apply quality control circle (QCC) to reducing non-timely maintenance rate of medical equipment.Methods QCC was established,and some measures were taken to observe its effect in decreasing non-timely maintenance rate of medical equipment.Results The non-timely maintenance rate fell from 12.1％ to 4.5％,and 0.3 million Yuan was saved for the manpower cost.Conclusion QCC decreases the non-timely maintenance rate of medical equipment,increases the efficiency of the maintainer,shortens the downtime of the fault medical equipment,enhances hospital economic benefit and patient satisfaction and provides references for other hospitals.

Objective To investigate the efficacy,adverse effect and the incidence of cardiovascular and cerebrovascular events by comparative study on simvastatin combined with ezetimibe on therapy of lowing blood lipids in metabolic syndrome patients.Methods One hundred and twenty metabolic syndrome patients were divided into experiment group (n=60) and control group (n=60).The experiment group was treated with 20 mg of simvastatin associated 10 mg of ezetimibe daily.The control group was treated with conventional-dose 20 mg of simvastatin.The period of treatment was 4 years in both groups.The efficacy,adverse effect and the occurrence of cardiovascular and cerebrovascular events were observed and analyzed.Results Blood lipid were significantly decreased in both groups after the treatment.There was significant difference on total cholesterol ( TC ) [ ( 3.05±1.17 ) mmol/L vs (4.94 ± 1.13 ) mmol/L,P ＜ 0.05 ],low density lipoprotein cholesterol ( LDLC) [ (2.12±1.03) mmol/L vs (3.16 ± 1.11 ) mmol/L,P ＜0.05] and triglyceride(TC) (1.02±0.29) mmol/Lvs (1.65 ± 0.32)mmol/L,P ＜ 0.05 ) between experiment group and control group.There were 3 cases of cardiovascular events in experiment group and 10 cases in control group,5 cases of cerebrovascular events in experiment group and 13 cases in control group.The incidence of cardiovascular and cerebrovascular in experiment group was significantly lower than that in control group (x2 =4.08 and 4.37,respectively,P ＜ 0.05 ).Conclusion Combination therapy with simvastatin and ezetimibe is more effective and it can significantly decrease the incidence of cardiovascular and cerebrovascular events in metabolic syndrome patients.

Objective To construct two kinds of eukaryotic ccll expression vcctors pIRES2-EGFP-SP-B-C/T 1580 and evaluate their expressions in 293T cells,for the further study of relationship between polymorphism of surfactant protein B (SP-B) gene and bronchopulmonary dysplasia (BPD).Methods The eukaryotic pIRES2-EGFP-SP-B-C/T 1580 expression vectors were constructed by gene recombination,and identified by gene sequencing.The recombinant expression vectors were transfected into 293T cells by lipofectamine2000.The expression of green fluorescence protein in 293T cells was observed by fluorescence microscopy.The mRNAs and proteins of SP-B-C/T 1580 were tested and identified by reverse transcriptionpolymerase chain reaction-restriction fragment length polymorphism(RT-PCR-RFLP) and western blot.Results Two recombinant plasmids contained the complete cDNA of SP-B with the same sequence as in gene bank.The base of SP-B 1580 gene of pIRES2-EGFP-SP-B-C 1580 was C,that of pIRES2-EGFP-SP-B-T 1580 was T.After being transfected into 293T cells,highly efficient expression of SP-B-C/T 1580 gene was detected at mRNA and protein levels.Conclusions The pIRES2-EGFP-SP-B-C/T 1580 eukaryotic cell expression vectors were successfully constructed.