For the purpose of determining the time of death,the life history (the growth and development) and the sum of effective temperature of Lucilia Sericata were studied under the ordinary temperature. On the bases of the growth and development of the fly ,we have established three linear regression equations between the sum of effective temperature and the length of larvae or weigth pupa of Lucilia Sericata (K 1=2 0880+0 8014X 1、K 2=54 0917-2 8814X 2、K 3=133 2180-2 6312X 3).These linear regression equations provide a reliable,scientific method for estimating the time of death in forensic medicine practice.

There were 48 strains of bacilli obtained from 20 Chinese traditional medicines. Twenty-five strains had antagonistic effect against at least one of ten plant pathogens. Seven strains had antibiosis to more than four pathogens and the best strain had antibiosis to nine pathogens. After physiological and biochemical experiments,eight strains of 25 antagonistic bacilli were proved to be Bacillus subtilis,three were Bacillus cereus,one were Bacillus natto and one were Bacillus licheniformis. At the same time,two kinds of Chinese traditional medicines,which probably had antibacterial effect,were found.

Objective To study the association of peptidylarginine deiminase 4 (PADI4) (padi4_94 and padi4_104) genetic single nucleotide polymorphism (SNP) and rheumatoid arthritis (RA) susceptibility in Han population of Hebei province. Methods This hospital-based case-control study included 115 RA patients and 106 healthy controls. All the individuals were recruited from Han Hebei residents and were randomly selected. The genotype and allele frequencies of PADI4 gene polymorphisms (padi4_94 and padi4_104) were analyzed by PCR-DNA sequencing method. Results The distribution of padi4_94 and padi4_104 genotypes between the two groups was not significantly deviated from that expected by Hardy-Weinberg equilibrium (P>0.05). The combined effect of padi4_94 and padi4_104 SNPs was analyzed and three haplotypos (AA, AG and GG) were found but without GA haplotype. The genotype and allele distribution of padi4_94 and padi4_104 in the patients with RA was not significantly different from that in healthy controls,and the analysis of haplotypes revealed that no haplotypes were risk factors for RA. Conclusion In Han population of Hebei province, the SNPs of PADI4 (padi4_94 or/and padi4_104) is no associated with RA susceptibility. Therefore, it should not be regarded as a genetic risk factor for RA.

AIM: To explore the effect of pulmonary arterial smooth muscle cell (PASMC) apoptosis on the reversal of hypoxic pulmonary arterial remodeling during reoxygenation and its possible mechanism.METHODS: Male SD rats (n=24) were randomly divided into normoxia for 4 weeks group, hypoxia for 4 weeks group, reoxygenation for 1 week after hypoxia for 4 weeks group and reoxygenation for 6 weeks after hypoxia for 4 weeks group.Right ventricular systolic pressure (RVSP), right ventricular hypertrophy index, pulmonary arterial medial thickness (MT) and medial area (MA) as well as autophagy and apoptosis in the pulmonary arterial medial layer were examined during hypoxia-reoxygenation.The rat primary PASMCs were divided into normoxia for 48 h group, hypoxia for 48 h group, reoxygenation for 24 h after hypoxia for 48 h group and normoxia for 72 h group to explore the changes of PASMC autophagy and apoptosis following hypoxia-reoxygenation.Finally, primary PASMCs were divided into normoxia for 72 h group, reoxygenation for 24 h after hypoxia for 48 h group and reoxygenation for 24 h after hypoxia for 48 h + chloroquine (inhibitor of autophagy) group to investigate the effect of PASMC autophagy during hypoxia on the apoptosis during reoxygenation.RESULTS: After hypoxia for 4 weeks, the RVSP, during right ventricular hypertrophy index, MT and MA increased significantly compared with normoxia group (P<0.05), and gradually decreased during reoxygenation.The expression of LC3 in the pulmonary arterial medial layer increased evidently after hypoxia and gradually reversed during reoxygenation.Moreover, the P62 and cleaved caspase-3 expression decreased after hypoxia compared with normoxia group, and increased markedly following reoxyge-nation.The expression of cleaved caspase-3/PARP in rat primary PASMCs decreased significantly under hypoxia (P<0.05), and increased evidently during reoxygenation.The expression of P62 and LC3-II decreased markedly under hypoxia (P<0.05).After inhibition of PASMC autophagy under hypoxia, the expression of cleaved caspase-3/PARP decreased remarkably during reoxygenation (P<0.05).CONCLUSION: The PASMC apoptosis participates in the reversal of hypoxic pulmonary arterial remodeling, and the PASMC autophagy under hypoxia might facilitate its apoptosis during reoxygenation.

Objective To construct plvx-cyclooxygenase-2(COX-2)-DsRed and establish breast cancer cell line MCF7 which overexpressed COX-2, to explore the effect of COX-2 on breast cancer cell.Methods The full-length COX-2 PCR product was obtained by total COX-2 PCR primers and COX-2 cDNA vector.After the PCR product and lentiviral vector plvx-DsRed-Monomer-N1 were cut simultaneously by restriction enzyme BamH1 and Xholl, they were connected and sequenced, to get lentiviral vector plvx-COX-2-DsRed.After selected by puromycin, overexpressed COX-2 breast cancer cell line MCF7-plvx-COX-2-DsRed was obtained.The stable cell line was verified by real time PCR and Western blot.The differences of proliferation ability between stable cell line and normal one were compared by colony formation test and Western blot.Results The lentiviral vector plvx-COX-2-DsRed and stable cell line MCF7-plvx-COX-2-DsRed after selecting were obtained.COX-2 expression level of the stable cell line was 75.29 times as high as that of MCF7, and 64.91 times as high as that of cell line MCF7-plvx-DsRed-Monomer-N1 by PCR assay (P ＜ 0.05), which was consistent with the results of Western blot and microscope photo.MTT results showed that cell line MCF7-plvx-COX-2-DsRed had grown faster than cell line MCF7 and MCF7-plvx-DsRed-Monomer-N1 from the 2nd day (P ＜ 0.05), which was accordant with colony formation assay.MCF7-plvx-COX-2-DsRed cell line had higher c-myc expression and lower β-catenin expression than MCF7 cell and cell line MCF7-plvx-DsRed-Monomer-N1 detected by Western blot relative quantification (P ＜ 0.05).Conclusion The plvx-COX-2-DsRed lentiviral vector and cell line MCF7-plvx-COX-2-DsRed are successfully constructed.COX-2 can increase proliferation of MCF7 cells through up-regulating the expression of c-myc.

Background Increase in ophthalmic optical medical instruments and microsurgical applications leads to retinal photochemical damage and other problems delivery of a variety of devices,so the in-depth study and understanding of its pathogenesis after retina light damage can provide a reference for the clinical treatment of related diseases.Objective This study was to investigate the therapeutic effect and relative mechanism of erythropoietin (EPO) on mouse retina photic injury by studying the expression of matrix metalloproteinases-2 (MMP-2)and MMP-9.Methods Fifty-two SPF BALB/c mice were randomized into normal control group,simple light-induced group and EPO pretreatment group by balloting method.The mice of simple light-induced group and EPO pretreatment group were continuously irradiated with 6000 lx diffuse light for 4 hours in a home-made box to establish the models of light-induced damage;while recombinant human EPO (rhEPO)of 5000 U/kg was intraperitoneally injected prior to the light exposure in the EPO pretreatment group.The expressions of MMP-2 and MMP-9 were examined at 6,12,36,72,96 hours and 7 days following light-exposure by immunohistochemistry.Results Edema and structural disorder of RPE cells appeared inthe simple light-induced group after light-exposure and aggravated with lapse of light-exposure time,but no similar change was seen until 7 days in the EPO pretreatment group.The immunohistochemistry findings showed that the expression of MMP-2(A value)in RPE cells was less in the normal mice.However,a large quantity of positive cells appeared in RPE layer 36 hours after light-exposure.Compared with the simple light-induced group,the positive expression of MMP-2 protein in EPO pretreatment group was significantly decreased,showing statistically significant differences among these three groups and different time points (Fgroup =3.68,P =0 04; Ftime =9.13,P=0.00).There was hardly any MMP-9 expression in the retina of the normal mice.In simple light-induced group,a few of positive cells appeared in RPE layer 6 hours after light-exposure and reached its peak 12 hours following light-exposure.The gradually down-regulation of MMP-9 expression happened 96 hours later following light-irradiation.The expression tendency of MMP-9 in EPO pretreatment group was similar to the simple light-induced group.Significant differences in expressions of MMP-9 were found among different groups and time points (Fgroup=3.61,P =0.04;Ftime =16.91,P=0.00).Conclusions MMP-2 and MMP-9 may be involved in the mechanism of retina photic injury by down-regulating the expression of MMP-2 and MMP-9 in RPE cells.

Objective To detect the levels of CD4+CD25HiCD127Low regulatory T cells (Treg) in the peripheral blood and its clinical significance in patients with esophageal cancer. Methods The levels of Treg in the peripheral blood were detected by three-color flow cytometry (FCM) in 80 patients with esophageal cancer and 20 healthy controls. Among the 80 patients, 30 patients were also further studied for preoperative and postoperative comparison after operation. The clinical and pathological data of each patient were collected and analyzed for the correlation with the Treg levels. Results The level of Treg in the peripheral blood of the control group was lower than that of the esophageal cancer patients [(3.36±1.14) % and (5.70±1.96) %, respectively], with significant difference (P <0.01). The levels of Treg in the peripheral blood was higher in the patients with metastasis of lymph node (n=40) than that in the patients without metastasis of lymph node (5.96±1.36) % and (4.23±1.18) %, respectively] (n=30), with significant difference (P <0.01). The levels of Treg in the peripheral blood of the patients were negatively correlated with their TNM classification. As the TNM classification advanced, the level of the Treg in the peripheral blood increased. Conclusion The levels of Treg in the peripheral blood of the patients with esophageal cancer are significantly higher than that of the healthy subjects, which is correlated with the clinical and pathological conditions. The development of esophageal cancer may relate with suppression of immune function.

OBJECTIVE To find out the main commonly encountered pathogen and antibiotic resistance in hospital infection in Department of Neurosurgery. METHODS Clinic samples from Jun 2005 to Jun 2007 were collected and drug sensitive test was taken.RESULTS The most commonly encountered pathogens of infection were Gram-negative bacillI(83.1%),including Klebsiella pneumoniae(25.0%),Escherichia coli(19.1%),andAcinetobacter baumannii(10.3%). The three kinds of pathogens had heavy resistance to at least 8 kinds of antibiotics. The resistance rates to imipenem and ofloxacin were the lowest (6.8% and 32.4%). CONCLUSIONS The pathogens isolated from Departmentof Neurosurgery are Gram-negative bacilli which have multiple antibiotic resistance. The key prevention measures of infection are to control prophylatcic usage of the third generation cephalosporins,strengthen environmental microbial monitoring,hand sterilization and cleaning among the medical staff.

Objective To study the gene polymorphism at position padi4_94 in Chinese Han people of Hebei Province and the characters of distribution.Methods 106 healthy individuals and 115 rheumatoid arthritis patients in Hebei Province were randomly selected. Padi4_94 gene was screened by sequencing of the promotor. The correlations between the SNP site and anti-cyclic citrullinated peptide antibody (anti-CCP) was discussed.Results There were three genotypes: AA, AG and GG in Chinese Han people of Hebei Province. No significant differences of gene frequency and allele gene frequency were found between healthy individuals and RA patients, and no correlation between SNP and the production of anti-CCP.Conclusions Hebei Province The gene polymorphisms at position padi4_94 existed in Chinese Han people of Hebei Province,but possibly would not affect RA susceptibility and the production of anti-CCP.

Secondary glaucoma is a kind of complications after vitrectomy, its etiologies are various and complex. Ineffective therapies might cause irreversible damage on optic nerves and visual field defect, even the loss in visual function. Nowadays, this project has been paid great attention by various researches both in China and abroad. Both the pathogens and therapies of secondary glaucoma after vitrectomy are analyzed as follows.