Objective:To study the association between rs2235371,rs2013162,rs2235377 SNPs in interferon regulatory factor 6 (IRF6)gene and non-syndromic cleft lip with or without cleft palate(NSCL/P)in Xinjiang Uyghur population.Methods:100 Uyghur NSCL/P patients from Xinjiang were included in the case group and 60 Uyghur inpatients with upper respiratory tract infection were se-lected in the control group.Next,generation sequencing was used,DNA sequencing results were compared with the information on the genome database and genetic analysis were made.Results:There were no significant differences in the frequency distribution of both genotypes and alles when the cases were campared with the controls at the rs2235371,rs2013162 and rs2235377 loci(P >0.05). Above three loci were located in the same block,rs2235371 and rs2235377 loci presents the strong linkage disequilibrium(r2 =0.949, D'=0.974).Possible haplotypes were:CCT >CAT >TAC,and there was no significant difference between the cases and controls in haplotype distribution(P >0.05).Conclusion:Polymorphisms of rs2235371,rs2013162 and rs2235377 in IRF6 gene may be associ-ated with NSCL/P in Xinjiang Uygur people.

OBJECTIVE: To investigate the anti-tumor effect of bufalin and its regulation on Bcl-2 and Bax proteins in orthotopically transplanted tumor of human hepatocellular carcinoma in nude mice. METHODS: Orthotopically transplanted tumor of human hepatocellular carcinoma was established in nude mice. The mice were randomly divided into five groups: high-dose bufalin-treated group (1.5 mg/kg), medium-dose bufalin-treated group (1 mg/kg), low-dose bufalin-treated group (0.5 mg/kg), adriamycin-treated group (8.0 mg/kg), and normal saline-treated group. After 25 days, mice were sacrificed. The tumor volume was measured, and the pathological changes of tumor tissues were detected by HE staining to observe the tumor necrosis degree. Cell morphological changes were also observed by an electron microscopy. Label index of tumor cell apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), and the expressions of Bcl-2 and Bax proteins were determined by immunohistochemical method. RESULTS: The tumor volume in the bufalin-treated groups was shrunk significantly compared with that in the normal saline-treated group (P<0.01). The survival time of the bufalin-treated groups was prolonged compared with that of the adriamycin-treated group and the normal saline-treated group P<0.05. Apoptotic characteristics could be seen in tumor tissues of the bufalin-treated groups. The label index of tumor cell apoptosis in the bufalin-treated groups (5.87+/-2.13, 8.86+/-2.96 and 10.60+/-3.42 in low-, medium- and high-dose groups respectively) was higher than that in the adriamycin-treated group (3.28+/-0.98) (P<0.05, P<0.01). The expression of Bax was up-regulated, while no changes were detected as to Bcl-2 protein in tumors of the bufalin-treated groups. CONCLUSION: Bufalin has significant anti-tumor effect on the orthotopically transplanted tumor of human hepatocellular carcinoma in nude mice. Its effect might be related to up-regulation of Bax protein and inducement of the tumor cell apoptosis.

Objective To study the reversal effect on acquired multidrug resistance (MDR) of human hepatocellular caricinoma (HCC) by Ganli Injection. Methods Proliferative inhibition rate of HCC BEL-7402/5-Fu cell line and reversal index of 5-Fu acquired MDR were measured by MTT assay. Cell morphological changes were observed under inverted microscope and Wright-Giemsa stain method. Fluorescence intensity of Adriamycin (ADM) was detected by Flow cytometry. Results HCC BEL-7402/5-Fu cell line exhibited cross-resistance to general chemotherapeutics in clinic. Proliferative inhibition rates of HCC BEL-7402/5-Fu cell line treated for 72 h by Ganli Injection at the (matrine) concentration of 0.230, 0.115, 0.058 mg/mL were (9.25?2.38)%, (8.46?1.90)%, and (4.23?2.05)%, respectively. The corresponding reversal indexes to 5-Fu were 5.50, 3.83, and 2.25. Pre-treatment of Galli Injection at the concentration of 0.230 mg/mL for 2 h could increase the accumulation of ADM in cells to 38.61%.Conclusion Ganli Injection has the effect on reversing 5-Fu acquired MDR of human hepatocellular carcinoma in vitro.

Purpose:To study the relation between regional lymph node metastasis and nm23 H 1 protein expression and DNA ploidy of esophageal squamous cell carcinoma patients. Methods:Flow cytometry was used to analyze nm23 H 1 protein and DNA ploidy in surgical specimens from the patients with esophageal carcinoma. The object tissues included: the primary tumor tissue, the pericancerous mucosa, the incisional margin and the regional lymph node. Results:There were obvious differences between nm23 H 1 protein expression in lymph node metastasis positive and negative groups, primary tumor tissue and pericancerous mucosa, cancer tissue and the incisal margin( P 0.05).The differences of DI were distinct between cancer tissue and the pericancerous mucosa, cancer tissue and the incisal margin, cancer tissue and the lymph node( P 0.05).In corresponding tissues of lymph node metastasis positive and negative groups, nm23 H 1 protein expression was different( P 0.05). As to DI, it was different only among pathological grades( P 0.05). Conclusions:nm23 H 1 protein higher expression can produce certain inhibitory effect on metastasis of esophageal carcinoma. Combining DNA ploidy detection can help to understand the biological behavious and deduce the prognosis of esophageal carcinoma.

OBJECTIVE: To study the feasibility of the establishment of the orthotopic transplantation tumor model of hepatocellular carcinoma in mice and its tumor biological characteristics. METHODS: H22 cells of hepatocellular carcinoma were inoculated to form ectopic transplanted model in mice by subcutaneous injection. Then the subcutaneous tumors were implanted into the liver of mice, and the orthotopic transplantation tumor model of hepatocellular carcinoma was established. RESULTS: The successful rate of the orthotopic transplantation tumor model was 95.6% and the spontaneous metastatic rate was 81.8%, the rate of mass ascites was 40.9% and the natural extinctive rate was 0%. The natural survival time in the orthotopic transplantation tumor model was 28 days and the proliferation of tumor in transplanted model was accelerated after 2 weeks or so. CONCLUSION: The orthotopic transplantation tumor model in mice is an ideal model for studying the metastatic mechanism and screening anti-tumor drugs for liver cancer, just because of its high successful rate and high spontaneous metastatic rate with no natural extinction.

AlM: To monitor long - term changes of matrix metalloproteinase-2 ( MMP-2 ) in human tears fluid after laser in situ keratomileusis ( LASlK) .
METHODS: Thirty - two myopia cases ( 64 eyes ) underwent uneventful LASlK were enrolled in the study. Tear fluid were collected and MMP-2 expression was analyzed by Western - bolt assay preoperatively and postoperatively on 15d, at 1, 3mo, and 1a.
RESULTS:LASlK increased the concentration of MMP-2 in human tear fluid. At 15d postoperatively, the magnitude of MMP-2 was 1. 4 times that of preoperative, thereafter subsided, but didn't return to preoperative level by 3mo ( P < 0. 05 ). Up to 1a after surgery, the concentration of MMP-2 almost recovered (P>0. 05).
CONCLUSlON: MMP- 2 is significantly expressed in human tear fluid after LASlK, then subsided with time, but didn't return to preoperative level by 3mo and almost recovered up to 1a, indicating wound healing of LASlK would continue up at least 3mo after surgery and almost recovered 1a postoperatively.

To study the clinical efficacy of Pihui Fanggan Sachet (PHFGS), a sachet of traditional Chinese herbs, in preventing influenza and its immune regulation on mice.

OBJECTIVE: To establish a multidrug-resistant cell line BEL-7402/5-FU of hepatocellular carcinoma (HCC). METHODS: BEL-7402/5-FU was induced by pulse therapy combined with continuous stepwise exposure to 5-fluorouracil in vitro. MTT assay was used to determine its multidrug resistance (MDR). Biological characteristics of the BEL-7402/5-FU cell line were observed including morphological changes, cell growth curve, population doubling time, plate cloning efficiency, adherence rate, cell cycle distribution, chromosome and tumorigenicity. Accumulation amount of adriamycin (ADM) in cytoplasm was measured by flow cytometry. The protein expression of thymidylate synthase (TS) was evaluated by immuno-cytochemical method. RESULTS: The acquired MDR cell line of BEL-7402/5-FU was established successfully. The BEL-7402/5-FU cells showed cross-resistance to ADM, vincristine (VCR), methotrexate (MTX) and oxaliplatin (OHP), whereas still sensitive to hydroxycamptothecin (HCPT). The BEL-7402/5-FU cells tended to grow in clusters in vitro. It was found that the population doubling time of BEL-7402/5-FU cells was longer than that of its parental cells. The plate cloning efficiency and the adherence rate of BEL-7402/5-FU cells at the 2nd and 3rd hour were both lower than those of the parental cells. The distributing proportion of BEL-7402/5-FU cells in G(0)/G(1) phase was less than that of the parental cells, whereas the distributing proportion of BEL-7402/5-FU cells in S phase was higher than that of the parental cells. The accumulation amount of ADM in cytoplasm of BEL-7402/5-FU cells was significantly lower while the expression level of TS protein of which was highly up-regulated as compared with those of the parental cells. CONCLUSION: Establishment of the human HCC cell line BEL-7402/5-FU might be beneficial to the studies of 5-Fluorouracil acquired MDR mechanisms and the selection of reversal modifiers.

Objective To breed homozygous mice of Treg-Th17 fluorescence double labeling and to investigate the role of Treg-Th17 balance in endotoxemia. Methods Mice of Treg and Th17 fluorescence single labeling were brought from Harvard Medical School,USA and were mated and bred based on the genetic rules.The genomic DNA was extracted from the tails of second generation weaning mice for genotyping by polymerase chain reaction (PCR).Thereafter,the homozygous mice of fluorescence double labeling were selected to have successive inbreeding for three generations and their genotypes and growth status were measured.The endotoxemia model in vivo was duplicated and changes of Treg-Th17 balance were detected by flow cytometry analysis at 24 h and 48 h after endotoxemia. Results Among the seven second generation mice in the first batch,only three (two males and one female) were the wanted homozygotes.Meanwhile,the genotypes of their offsprings for the next three generations were all Foxp3 RFPKI-homo IL-17A GFPKI with normal growing status.As important component of endotoxin,lipopolysaccharide (LPS)could induce significant signal expressions of red fluorescence protein (RFP) and green fluorescence protein (GFP).The ratio of CD8a + Foxp3 +,CD8a + IL-17A +,CD4 + Foxp3 + and CD4 + IL-17A + in the lymphocytes went up significantly in the group of 24 hours of LPS treatment,as compared with the control group (P ＜ 0.01 or P ＜ 0.05 ),while the difference between the control group and the group of 48hours of LPS treatment had no statistical significance (P ＞ 0.05). Conclusion Foxp3 RFPKI-homoIL-17A GFPKI mice have identical genotypes and stable genetic characteristics and is a model animal and novel research platform that can provide real-time monitoring of the changes of Treg-Th17 balance in vivo.The role of T reg-Th17 balance works significantly at 48 hous after the subject is exposed to LPS stimulation,and the roles of CD8a + Foxp3 and CD8a + IL-17A + in Treg-Th17 balance require further investigation.

Objective: To evaluate the influence of recombinant human brain natriuretic peptide(rhBNP)on ventricular remodeling and ventricular systolic synchrony in patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI).Methods: A total of 48 patients with AMI after PCI were randomized into two groups: rhBNP Group (n=25) and Routine treatment Group (n=23).Two dimension echocardiography was used to measure the index of left ventricular end-diastolic volume (LVEDVI), the index of left ventricular end-systolic volume (LVESVI), left ventricular ejection fraction (LVEF), the index of left ventricular mass (LVMI), the movement index of infarcted regional wall (RWMI) at 1 week, 4 weeks and 24 weeks after the treatment procedure.And at 1 week and 24 weeks period, equilibrium radionuclide angiography was performed respectively to evaluate the ventricular systolic synchrony.Results: ①1 week after PCI, LVESVI in rhBNP group was decreased and LVEF was increased than that in Routine treatment group (P<0.05, respectively).4 and 24 weeks after PCI, LVEDVI, LVESVI, LVEF were significantly different in rhBNP group than those in Routine treatment group (P<0.05,respectively).24 weeks after PCI, RWMI and LVMI were significantly decreased in rhBNP group than those in Routine treatment group(P<0.05,respectively).②24 weeks after PCI, phase shift, full width at half maximum and peak phase standard deviation were significantly improved in rhBNP group than those in Routine treatment group (P<0.05, respectively).Conclusion: Intravenous injection of rhBNP could inhibit left ventricular remodeling and improve left ventricular function.