Objective To investigate relationship of platelet (PLT) count with clinicopathological features and prognosis of patients with breast cancer,and explore the susceptibility index to evaluate prognosis of patients with breast cancer.Methods A retrospective analysis of clinical data of 498 patients with breast cancer in January 1995 to December 2005 was carried out.PLT count was tested.Those patients were divided into group A (PLT ＜ 150 × 109/L),group B[(150-250) × 109/L],and group C (PLT ＞ 250 × 109/L) according to PLT count level.The relationship of platelet count with clinicopathological features was analyzed.Kaplan-Meier and Cox proportional hazards model were used for univariate analysis and multivariate analysis of PLT impact on survival time.Results There was positively correlated between PLT count and clinicopathological features (Pearson coefficient ＞ 0,P ＜ 0.05).There was negative correlated between PLT count and survival time (Pearson coefficient =-O.583,P ＜ 0.05).The survival time of groups A,B and C were significantly different (P =0.018).Cox proportional hazards model multi-factor analysis showed that PLT count was an independent factors affecting survival time (OR =2.256,P ＜ 0.05).Conclusions Patients with breast cancer associated with increased emphasis and PLT count.PLT count had negative correlation with survival time.PLT count could be a susceptible index to predict the prognosis of patients with breast cancer.

Objective To detect the impact of Diltiazem on the major adverse cardiac events (MACE) in six months after percutaneous coronary intervention (PCI).Methods A total of 192 patients after PCI with coronary atherosclerotic heart disease were enrolled in this study.The patients were randomly divided into Diltiazem therapy group (101 patients) and non-Diltiazem therapy group (91 patients).The high-sensitivity C-reactive protein (hs-CRP) was assessed before and 24 h after PCI,and the incidence of Major adverse cardiovascular events(MACEs) were assessed at the sixth month after PCI.Results Compared with before PCI,hs-CRP level increased significantly in both group after PCI (P＜0.01),but hs-CRP level was lower in Diltiazem therapy group than in non-Diltiazem therapy group (P＜0.05).Compared with non-Diltiazem therapy group,there was lower incidence of MACEs during six months follow-up in Diltiazem therapy group.Conclusions Diltiazem can decrease the incidence of MACEs during six months after PCI.

Objective To analyze the factors correlated with perinatal death so as to provide the basis for reducing the perinatal mortality.Methods A respective study was conducted with analysis of perinatal mortality monitoring data,birth defects data and the health status report of non-registered pregnant women from 2012 to 2016 in Taizhou. We compared the differences in the indexes of perinatal mortality,birth defects rate and the proportion of elderly pregnant women in different years,domicile place and regions.Results The perinatal mortality rate was 6.80‰,decreasing annually from 2012 to 2016 (P<0.01). The average perinatal mortality rate of floating population was 9.28‰,which is higher than the 5.64‰ rate of local population (P<0.01). The proportion of elderly pregnant women was 10.30%,showing an upward trend (P<0.01) and the perinatal mortality of elderly pregnant women was 10.60‰,significantly higher than the total mortality (P<0.01). The leading cause of perinatal death was birth defect and the defect rate of perinatal birth was 35.86‰,showing an upward trend (P<0.01) while the average mortality rate of birth defects was decreasing (P<0.01). There were statistically significant differences(P<0.01) in perinatal mortality,birth defects,sex ratio and proportion of elderly pregnant women in different regions of Taizhou. Compared with the perinatal in local population,mother age,education background,maternal times and register time of pregnant women of the perinatal in floating population was significantly different (P<0.01). Conclusion The perinatal mortality in Taizhou declined year by year. Elderly pregnant age,birth defects,and floating population are the main positive factors of perinatal mortality.

This study was purposed to investigate the therapeutic effects of early transfusion of immunized donor lymphocytes after haploidentical transplantation by means of mouse model of nonmyeloablative haploidentical bone marrow transplantation. CB6F1 female mouse was served as recipient and C57BL/6 male mouse was served as donor. Each CB6F1 female mouse was subjected to intravenous transfusion with 1×10(6) erythroleukemia (EL9611) cells at day 4 before transplantation, followed with intraperitoneal injection of Ara-C (0.015 g) respectively at day 2 and day 1, then conditioned for BMT with TBI (450 cGy) at day 1 before transplantation. After conditioning (day 0), each of recipients was transplanted with 6×10(7) mixture of bone marrow and spleen cells from the C57BL/6 mice, and was infused with 6 × 10(7) immunized donor lymphocytes at day 15 after transplantation. All treated animals were evaluated for survival, development of leukemia and aGVHD. The donor CD3(+) cell chimerism and sex determining region Y gene (SRY)in recipients were monitored periodically after transplantation. The results showed tht all mice with only inoculation of 10(6) EL9611 cells survived for 15 ± 1 days (n = 4); all mice of other groups obtained the varying degrees of implantation. SRY could be detected at day 30 and 60 after transplantation. The chimerism of donor CD3(+) cells in mixed bone marrow transplantation (MT) group at day 14, 30 and 60 respectively reached 17.95% ± 12.03%, 37.34% ± 2.78% and 47.06% ± 6.1%. In donor lymphocyte infusion (DLI) group it reached 69.78% ± 12.62%, 75% ± 15.97%, 83.41% ± 16.07% at day 30, 45 and 60 after transplantation. The mice of MT and DLI group survived for 66.66 ± 1.47 days and 78.2 ± 7.82 days. It is concluded that the high tumor burden before transplantation can affect donor cell engraftment and prognosis.Early post-transplanted infusion of immunized lymphocytes from donor can help to improve the therapeutic efficacy and survival.
Animals ; Bone Marrow Transplantation ; methods ; Female ; Haplotypes ; Leukemia, Erythroblastic, Acute ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation Conditioning ; methods ; Transplantation, Homologous

OBJECTIVETo investigate HIV infection status and its influence factors in men who have sex with men (MSM) in Sichuan province.

METHODSA face to face investigation and Syphilis and HIV serological detection were conducted among 2 783 MSM who have the insertion of oral or anal sex with men over the past year recruited by snowball sampling and respondent driven sampling methods in nine cities' Gay venues of Sichuan province. χ(2) test and logistic regression analysis were used for statistical analysis.

RESULTSA total of 2 783 MSM were surveyed, of which HIV and Syphilis infection rates were 11.1% (147/2 783) , 5.3% (310/2 783) , respectively. The HIV infection rates of <20 year-old age group, 20-29 year-old age group, 30-39 year-old age group, 40-49 year-old age group, ≥ 50 year-old age group were 6.6% (15/227), 9.7% (143/1 471), 11.8% (80/679), 18.0% (53/294), 17.0% (19/112) (χ(2) = 25.91, P < 0.05). The risk of HIV infection in 30-39 year-old age group, 40-50 year-old age group,> 50 years age group were 2.05 (1.14-3.69) times, 3.24 (1.75-6.01) times, 2.60 (1.22-5.52) times respectively of the <20 years age group. The risk of HIV infection in middle school and below one was 16.5% (73/443) , higher than the high school/college education MSM (11.1% (99/891) ) and the college and higher education MSM (9.5% (138/1 446)) (χ(2) = 16.46, P < 0.05). The risk of HIV infection in High school/college education MSM were 0.64 (0.45-0.90) times of the middle school and below. The HIV infection rates of MSM who accepted a HIV test and knew the result within the last year was 8.2% (119/1 446) , lower than the group who did not accepted any HIV test (14.3% (191/1 336) ) (χ(2) = 25.81, P < 0.05). The HIV infection rates of MSM who received intervention services was 10.1% (256/2 539), lower than the group who did not receive any intervention services (22.1% (54/244) ) (χ(2) = 32.65, P < 0.05) . The HIV infection rates of Syphilis-positive MSM was 32.0% (47/147) , higher than the Syphilis-negative one (10.0% (263/2 636) ) (χ(2) = 68.06, P < 0.05). Received intervention services (OR (95%CI) was 0.52 (0.40-0.68) ) and accepted a HIV test and knew the result within the last year (OR (95%CI) was 0.52 (0.36-0.74) ) were the protective factors of HIV infection. At the same time, the syphilis infection (OR (95%CI) was 4.01 (2.73-5.88) ) were risk factors for HIV infection.

CONCLUSIONThe prevalence rates of HIV infections were considered to be high among MSM in Sichuan province. The MSM of low-literacy, 30 years or older, not received any intervention services, not received any intervention services.Syphilis-positive have a greater risk of HIV infection.

Adolescent ; Adult ; China ; Data Collection ; Demography ; HIV Infections ; HIV Seropositivity ; Homosexuality, Male ; Humans ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Sexual Behavior ; Surveys and Questionnaires ; Syphilis

OBJECTIVETo study the effects of intestinal microflora alteration on specific and nonspecific immune function and hematopoietic function of mice.

METHODSSixty BALB/C mice were divided at random into two groups, experimental group and control group, with 30 mice in each. The mice in the experimental group were given kanamycin 50 mg while those in the control group were given distilled water intragastrically everyday for consecutive 10 days. After the 10 day treatment all the mice were sacrificed, and the cecal contents were collected for quantitative analysis of the intestinal bacterial flora. Certain indexes of immune function, including phagocytosis rate of macrophages, number of T lymphocytes positively stained by esterase and serum interleukin 2 (IL-2) content, and the weight of the spleen, granulocyte-macrophage colony stimulating factor etc. as indexes of hematopoietic function were determined.

RESULTSIn the group, the quantity of Enterobacteriaceae, Enterococcus, Bifidobacterium and Lactobacillus were significantly lower than that in the control group (P < 0.01). The number of PFC (plaque forming cells), the phagocytosis rate of macrophage, the number of T lymphocytes with positive NANE staining, the level of IL-2 significantly decreased when compared with that in the control group (P < 0.01). The weight of the spleen in the experimental group decreased when compared with that in the control group (P < 0.01). Levels of IL-3, GM-CSF, the total number of WBC and the proportion of neutrophil remarkably decreased as compared to that in the control group (P < 0.01). Analysis of the correlations between normal microflora, immunologic and hematopoietic indexes showed that marked positive correlations between the quantity of Bifidobacteria and each immune index including the levels of IL-3 and GM-CSF. There was a positive correlation between IL-2 and IL-3, IL-2 and GM-CSF as well.

CONCLUSIONThe application of antibiotics may cause changes in the structure and quantity of intestinal microflora. The dysbacteriosis may decrease the immune function of organism. The dysbacteriosis may decrease the hemopoietic function. The dysbacteriosis, the decrease in immune and hematopoietic function may affect one another. The balance in microecosystem should be emphasized and antibiotics should be applied rationally to reduce the side effects such as dysbacteriosis.

Animals ; Anti-Bacterial Agents ; pharmacology ; Esterases ; biosynthesis ; Feces ; microbiology ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; Interleukin-2 ; blood ; Intestines ; drug effects ; microbiology ; Kanamycin ; pharmacology ; Macrophages ; drug effects ; physiology ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Organ Size ; Phagocytosis ; drug effects ; Spleen ; drug effects ; pathology ; T-Lymphocytes ; drug effects ; metabolism

OBJECTIVETo investigate the changes of gene expression profile in transitional cell carcinoma of bladder T24 cell after crocin treatment, in order to find the possible crocin targets.

METHODThe bladder cancer T24 cell line was treated with crocin. MTT assay was adopted to determine the inhibition rate for selecting the best effect time and concentration of crocin. Differentially expressed genes on groups with or without treatment of crocin were screened with high throughout cDNA microarray. One up-regulated gene p21(WAF1) and one down-regulated gene cyclinD1 were selected to undergo analysis by the reverse transcription polymerase chain reaction (RT-PCR). Moreover, immunocytochemical method was used to evaluate p21(WAF1) and cyclinD1 protein expression.

RESULTThe growth of T24 cells was inhibited remarkably following a marked positive correlation between crocin concentration, time and inhibitor rate. When 3 mmol x L(-1) crocin treated T24 cells for 48h, the difference was significant compared with the control group (P < 0.05). Crocin induced wide changes of the gene expression profile of T24 cells. A total of 836 genes were up-regulated or down-regulated by more than 2 times, which were involved cell cycle controlling, DNA cell apoptosis, replication factor, and so on. The mRNA expression of p21(WAF1) and cyclinD1 detected by RT-PCR were in accordance with cDNA microarray data. The results of immunocytochemical method showed that p21(WAF1) and cyclinD1 protein expression were consistent with those mRNA expression.

CONCLUSIONCrocin can induce the significant alteration of gene expression profile of T24 cell. It is suggested that the widly konwn anti-tumor effects of crocin are medicated at least in part by regulating the cell cycle controlling gene expression.

Antineoplastic Agents ; pharmacology ; Carotenoids ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; drug effects ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immunohistochemistry ; Oligonucleotide Array Sequence Analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors ; Up-Regulation ; drug effects ; Urinary Bladder Neoplasms ; metabolism ; pathology

In order to improve the fermentation potency of phytase in recombinant host and decrease the production cost, the pichia expression vector pGAPZalpha-A was modified by introduction of an AOX1 promoter from vector pPIC9 and the resulted vector pAOXZalpha is an methanol induced vector. After that, a phytase gene appA-m was cloned into pAOXZalpha to construct the recombinant vector pAOXZalpha-appA-m. The recombinant Pichia pastoris 74#, which already contains one copy of appA-m and its fermentation potency exceeded 7.5 x 10(6) IU/mL, was used as the host strain for the transformation of pAOXZalpha-appA-m. The Pichia pastoris transformants were gained by electroporation. PCR results indicated that the appA-m expression box has integrated into the genome of Pichia pastoris and the original construction of phytase gene has not changed. SDS-PAGE analysis revealed that phytase was overexpressed and secreted into the medium supernatant. Recombinants with high expression level were screened and used for fermentation. In 5L fermentor, the expression level of phytase protein achieved 4 mg/mL and the phytase activity (fermentation potency) exceeded 1.2 x 10(7) IU/mL, which was about 1.6-fold compared with that of the host strain 74#. Moreover, the improved recombinant Pichia pastoris is excellent at expression stability and heredity stability.
6-Phytase ; genetics ; Fermentation ; Gene Dosage ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombination, Genetic

A gene appA encoding a novel phytase was firstly cloned from Hafnia alvei by PCR and sequenced. The gene was consisted of 1335 bp, encoding 444 amino acids. The calculated molecular weight of the mature APPA was about 45.2 kD. The gene appA was expressed in E. coli BL21 (DE3). Recombinant APPA was purified and its enzymatic properties were determined. The optimum pH for the enzyme was 4.5 and the optimum temperature was 60 degrees C. The pH stability of r-APPA is good, the relative phytase activity was above 80% after treated in buffers of pH 2.0-10.0. The specific activity of r-APPA is 356.7 U/mg, and the Km value was 0.49 mmol/L and Vmax of 238 U/mg. The enzyme showed resistance to pepsin and trypsin treatment.
6-Phytase ; biosynthesis ; genetics ; isolation & purification ; Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Hafnia ; enzymology ; genetics ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Temperature

OBJECTIVETo investigate the proliferation, apoptosis and mechanisms on T24 cell of transitional cell carcinoma of bladder (TCCB) by crocin.

METHODMTT assay was used to evaluate the proliferation of T24 cells. The changes of cell cycle and cell apoptotic percentage were measured by flow cytometry. T24 cells were inoculated into BALB/c nude mice to establish model of carcinoma of bladder. The mice were randomly divided into control group and experimental group. After treatment with 50 mmol x L(-1) crocin, the inhibited growth of tumor was observed. Electronic microscope was used to observe the morphological changes. The expressions of Bcl-2, Bax, Survivin and Cyclin D1 were detected by immunohistochemistry.

RESULTThe growth of T24 cells was remarkably inhibited after treatment of crocin. Flow cytometric profiles revealed that crocin led to the increase of the cells in G0/G1 phase, the percentage of cell apoptosis was also increased. Crocin could inhibit the growth of BALB/c xenograft tumor. The morphology changes of cell apoptosis were observed. Bcl-2, Cyclin D1 and survivin expressions determined by immunohistochemical staining were down-regulated after treatment with Bax expression up-regulated.

CONCLUSIONCrocin exerts both in vitro and in vivo anti-cancer effect on TCCB T24 cell line. The mechanisms may change tumour cell cycle and induce tumour cell apoptosis by down-regulating the expression of Bcl-2, Survivin, Cyclin D1 and up-regulating the expression of Bax.

Animals ; Apoptosis ; drug effects ; Carcinoma, Transitional Cell ; drug therapy ; pathology ; ultrastructure ; Carotenoids ; pharmacology ; therapeutic use ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Repressor Proteins ; Transplantation, Heterologous ; Urinary Bladder Neoplasms ; drug therapy ; pathology ; ultrastructure