Objective To perform mouse pMSV-Slfn5-GFP gene recombinant expression plasmid construction and gene structure analysis.Methods Total RNA was extracted from mouse liver and turned into cDNA by reverse transcription.Mouse Slfn5 coding sequence (CDS) fragment was amplified by PCR and connected with the pGEM-T Easy vector.The connected product was transferred the E.coli DH5a.The positive clones were selected for extracting plasmid,which was identified by double enzyme of restriction endonuclease Hpa Ⅰ and Xho Ⅰ.Then correct plasmid identified by enzyme digestion was sequenced by Macrogen USA.Then correct plasmid by sequencing was connected with pMSV-GFP by HindⅢ and Xbo Ⅰ,which was named as pMSV-Slfn5-GF.UCSC (http://genome.ucsc.Edu/) was used to analyze mouse Slfn5 and its family genomic structure.Slfn5 protein structural domain was determined by NCBI.Results Slfn5 full-length gene sequence was cloned into the expression vector pMSV-GFP,the fragment size was about 2.65 kb by enzyme digestion identification.The conservatism of AAA_4 protein domain in Slfn5 protein family was determined by phylogeny.fr.Conclusion Mouse full-length gene pMSV-Slfn5-GFP expression vector is successfully constructed.

BACKGROUND:Specific mechanism of stem cel transplantation for treatment of myocardial infarction has been not very clear. OBJECTIVE:To investigate the effect of H2O2 METHODS: Twenty-four C57BL/6 mice, 9 weeks old, were randomized into myocardial infarction group, embryonic stem cels group, H-pretreated mouse embryonic stem cels in vitro for treatment of myocardial infarction. 2O2 pretreatment group, with eight mice in each group, which were respectively injected 150 μL PBS, 150 μL embryonic stem cels (1×106), and 150 μL H2O2-pretreated embryonic stem cels (1×106 RESULTS AND CONCLUSION:Compared with the myocardial infarction group, the left ventricular end-diastolic diameter and left ventricular end-systolic inner diameter were significantly decreased in the embryonic stem cels ) at 1 week after modeling. At 3 weeks after transplantation, left ventricular end-diastolic diameter, left ventricular end-systolic inner diameter and left ventricular ejection fraction of mice in each group were measured by ultrasound. Myocardial colagen content was detected by Sirius red staining and the average colagen volume fraction was calculated. group and H2O2 pretreatment group, while the left ventricular ejection fraction was significantly increased (P < 0.05). Heart weight/body weight, left ventricular weight/body weight, and average colagen volume fraction were significantly lower in the embryonic stem cels group and H2O2 pretreatment group than the myocardial infarction group (P < 0.05). Therefore,in vitro H2O2 pretreatment enhances reduce the post-myocardial infarction myocardial fibrosis and greatly improve heart function.

OBJECTIVE:To observe the clinical efficacy and safety of herbs-partitioned moxibustion combined with curcumin in the treatment of Crohn's disease. METHODS:80 patients with mild and middle Crohn's disease were randomly divided into ob-servation group and control group with 40 patients in each group. Both groups were given western medicine treatment,such as sul-fasalazine;control group was additionally given curcumin,and observation group were given curcumin combined with herbs-parti-tioned moxibustion. 2 months later,Therapeutic efficacy,ADR and complications of 2 groups were observed after treatment. RE-SULTS:The total effective rate of control group and observation group were 87.50% and 95.00%,respectively;that of observa-tion group was better than that of control group,with statistical significance(P<0.05). The incidence of ADR in control group and observation group were 22.50% and 7.50% respectively;that of observation group was significantly lower than that of control group,with statistical significance(P<0.01). The incidence of complications in control group and observation group were 22.50%and 7.50%,respectively;that of observation group was significantly lower than that of control group,with statistical significance (P<0.01). CONCLUSIONS:The clinical efficacy of herbs-partitioned moxibustion combined with ccurcumin is effective in the treatment of Crohn's disease,and can reduce the incidence of ADR and complications.

Objective To study the feasibility of re-operation of gynecological laparoscopy.Methods Clinical records of 47 patients with gynecological diseases undergoing a re-laparoscopy from December 1995 December 2003 were analyzed retrospectively.Results The re-operations of laparoscopy were successfully accomplished in 46 patients(97.9%) while a conversion to open surgery was required in 1 patient.The re-operation time was 30~80 min(mean,50 min),and the in-hospital stay was 2.5~5 days(mean,3.5 days).No long-term complications were noted in 44 patients on a postoperative follow-up for(1~4) years.Out of 19 patients with infertility, 1 patient had a natural pregnancy and 6 patients were pregnant with the help of artificial reproductive techniques.Supportive medical treatment had been given in patients with endometriosis postoperatively,3 of whom experienced symptomatic improvements and 1 of whom had a natural pregnancy.Conclusions For patients with a history of laparoscopy,a re-operation of gynecological laparoscopy is safe and feasible.

Objective To develop a sensitive, specific, rapid and easy method for detecting E. coli O157 : H7 and other Shiga toxin-producing E. coli in food and clinical specimens. Methods A circular probe and capture probe specific for Shiga toxin-2 (stx2) gene had been synthesized and was used for determining the sensitivity of ramification amplification method (RAM). Different serotypes which contained stx2 gene, including an E. coli O157 : H7, an E. coli O46 : H38, an E. coli O111 : NM, an E. coli O22 : H8 and E. coli ATCC23716 (stx2 gene negative) were used for determining the specificity. All strains were detected by RAM to determine whether they contained stx2 gene. Real-time RAM was further studied to detect stx2 gene. Results The lowest number targets detected by RAM assay was 10 copies of stx2, indicating that RAM assay was as sensitive as conventional PCR. The result of specificity showed that different serotypes of strains were all positive for stx2 gene, while nonpathogenic E. coli ATCC23716 was negative. Among 32 isolates, 28 STEC isolates containing stx2 gene were positive by RAM assay, while others were negative. The RAM results were 100% in concordance with that of PCR. The real-time RAM results also showed that as many as 10 bacteria can be detected and the time of appearance of detectable signal was depended on the target concentration. Conclusion RAM assay can offer an alternative method for PCR to detect E. coli O157 : H7 and STEC in all types of specimens because of its simplicity and isothermal amplification.

Objective To compare serum protein fingerprint among Uygur patients with thyroid cancer and benign thyroid nodules, and Han patients with thyroid cancer, and to screen ethnic-specific protein markers of thyroid cancer. Methods Using the technology of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the protein expression profiles of Uygur and Han patients with thyroid cancer and Uygur patients with thyroid nodules were established and compared by image analysis software between two groups. Liquid chromatography-mass spectrometry/mass spectrometry coupling techniques ( LC-MS/MS) were used to identify differential protein. The protein′s name, cellular localization and functional classification were searched in Swissport database. Results There are 11 differentially expressed proteins by comparison of sera in Uygur patients with benign thyroid nodules and thyroid cancer, in which complement C3 and C4b levels were down-regulated in the serums of Uygur patients with benign thyroid nodules, and 9 proteins such as heme-binding proteins etc were up-regulated. There are 7 differentially expressed proteins by comparison of serums in Uygur and Han patients with thyroid cancer, in which transferrin level was up-regulated in the serum of Uygur patients with thyroid cancer, and 6 proteins such as cytokeratin-1 etc were down-regulated in serum of Uygur patients with thyroid cancer. Conclusion Combined screening of multiple labelled proteins including heme-binding protein,α2-macroglobulin, and transferrin protein etc may provide the basis for the diagnosis of thyroid cancer in Uygurs and Hans.

Objective To investigate the relationship of nerve growth factor (NGF) between serum and follicular fluid in patients with polycystic ovary syndrome (PCOS), and its effect on pregnancy outcomes. Methods From December 2011 to November 2012, 65 PCOS patients suffered with in vitro fertilization-embryo transfer (IVF-ET) were included in PCOS group and 31 non-PCOS patients with IVF-ET were used as control group. The expressions of NGF in serum and follicular fluid were detected by ELISA on the injection day of chorionic gonadotropin (HCG). The expression levels of NGF in serum and follicular fluid were compared between PCOS group and control group. Results The NGF levels in serum[(14.38±0.42) ng/L] and follicular fluid[(9.61±0.49)ng/L] were significantly higher in PCOS group than those of control group[(11.39±0.38) ng/L and (7.55 ± 0.40)ng/L]. In PCOS group, NGF levels in serum [(14.22 ± 0.35)ng/L] and follicular fluid [(9.30 ± 0.31)ng/L] were significantly lower in pregnant group than those of non-pregnant group[(14.51±0.43)ng/L, (9.86±0.46)ng/L, P<0.01]. Conclusion The increased levels of NGF in serum and follicular fluid of PCOS patients contribute to the pathogenesis of PCOS. The high expression of NGF in serum and follicular fluid may be harmful to pregnancy in PCOS patients.

Objective To establish the quality standard for Tianqi Granule.Methods Radix Notoginseng in Tianqi Granule was identified by TLC.The ginsenoside Rg1 and notoginsenoide R1 in Tianqi were determined by HPLC.Result Radix Notoginseng could be identified by TLC.The linear ranges of ginsenoside Rg1 and notoginsenoide R1 were at 0.628～5.652 ?g(r =0.999 8)and 0.155～1.395 ?g(r =0.999 8),respectively.The average recoveries were 99.24%(RSD=0.84%)and 99.14%(RSD=1.13%).Conclusion The method is simple,accurate and with good reproducibility and high precision,and can be used for quality control of Tianqi Granule.

Objective To analyse the effect of berberine on D-dimer, fibrinogen (FIB) and interleukin-6(IL-6) in peripheral blood of ovarian cancer patients.Methods 130 cases selected from the hospital were diagnosed as unilateral ovarian cancer, treated by surgery and chemotherapy.The patients were divided into control group (25 cases) and experimental group (105 cases), and the experimental group were randomly divided into A, B, C three groups, 35 cases in each group.The experimental group received berberine hydrochloride tablets, orally, three times a day, on the basis of conventional chemotherapy.The experimental groups were divided into three subgroups according to the dosage: A group: 0.3 g, B group: 0.2 g, C group:0.1 g, 35 cases in each group, continuously treated for 2 weeks.The levels of D-dimer, FIB and IL-6 and quality of life score were detected before and after treatment.Results After treatment, D-dimer level in each subgroup of experimental group was lower than that in control group, respectively, and A group