This study is to investigate whether naked plasmid DNA can effectively transfect lung cancer related cells and express human kallistatin, an endogenous protein that inhibits angiogenesis and tumor growth, and to explore the biological activity of the low-level expressed kallistatin to lung cancer in vitro and in vivo. The plasmids were delivered with Lipofectamine 2000 to transfect various lung cancer related cells. Kal expression was determined by ELISA. The biological effects of Kal expression on proliferation, migration and apoptosis rate of the cells were examined. In subcutaneous NCI-H446 xenograft model, pKal was injected directly into tumors, the changes of CD34, Ki-67 and E-cadherin expression were detected with immunohistochemical assay, the tumor apoptosis was analyzed with TUNEL assay. Both the endothelial cell and lung cancer cells could express kallistatin after plasmid transfection. The proliferation and migration of human umbilical vein endothelial cells were inhibited, but the apoptosis rate was not affected. The proliferation rates of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited, and their apoptosis rates were enhanced, but different cells behaved differently. In subcutaneous NCI-H446 xenograft model, intratumor injection of pKal inhibited the growth of lung cancer by reducing angiogenesis and proliferation of tumor cells. In conclusion, this study demonstrated the efficacy of plasmid-mediated expression of kallistatin to lung cancer related cells, thus providing a basis for their clinical application in the treatment of lung cancer.
Animals ; Antigens, CD34 ; metabolism ; Apoptosis ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; Humans ; Ki-67 Antigen ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Plasmids ; Serpins ; genetics ; metabolism ; physiology ; Transfection ; Tumor Burden

Objective To analyze time trend of cancer during 1970-2005 in Shandong province so as to develop strategies for control and prevention of cancer at the community level. Methods Data was from 4 retrospective surveys regarding all causes of death during 1970-- 1974, 1985-1989, 1990- 1992 and 2004-2005, in Shandong province. Other than one set of data collected in 1985-1989 by Shandong province itself, the other 3 set of data were from the national surveys, in which the survey-point sampling of choice was based on data of 1970-1974 for assessing its representativeness. The observing indices would include standardized mortality and mortality. A join-point regression model was used to analyze the changing rate of tumor. Results The mortality rate of the entire tumor increased 143.15 percent in 2005 than in 1970. The changing slope of standardized rate of all tumors in the regression model showed that the inter-annual growth rate were 0.54 and 1.24 percent from 1970 to 1984 and from 1985 to 1992. The rate of increase since 1992 had been 0.18 percent. During 2004-2005, the main malignant cancers were lung, stomach, liver, esophageal, coiorectal, leukemia, breast and cervical cancer, in order. Lung cancer rose from the 4th ranking to the first while cervical cancer dropped from the fifth ranking to the 8th place. Esophageal cancer and cervical cancer were decreased annually while gastric cancer was increased in the early days but decreased later on. The rest of the cancers were on the rise year by year. Rates of lung and breast cancers were higher while gastric and esophageal cancers were lower seen in the urban than in rural areas. Conclusion In Shandong province, a marked increase was seen in the mortality rate of tumors in the past 35 years. Evidence showed that the spectrum of death among main malignant tumors had changed which might provide a scientific basis for the development of a community-based prevention and control program on cancer.

Antigens, Surface ; analysis ; Antiretroviral Therapy, Highly Active ; CD28 Antigens ; analysis ; CD56 Antigen ; analysis ; HIV Infections ; drug therapy ; immunology ; Humans ; Killer Cells, Natural ; immunology ; Lectins, C-Type ; analysis ; NK Cell Lectin-Like Receptor Subfamily B ; NK Cell Lectin-Like Receptor Subfamily D ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR

BACKGROUNDAt the end of 2005, 650,000 people lived with human immunodeficiency virus type-1 (HIV-1) in China, of whom 75 000 were AIDS patients. Many AIDS patients received highly active antiretroviral therapy (HAART) supported by the "China CARES" program but the immune responses of HAART were seldom reported. This study investigated the effect of HAART on the activation and coreceptor expression of T lymphocytes in Chinese HIV/AIDS patients and evaluated its effect on immune reconstitution.

METHODSSeventeen HIV/AIDS patients were enrolled and three-color-flow cytometry was used to detect the activation of HLA-DR CD38 and the coreceptor CCR5, CXCR4 expression on T lymphocytes in whole blood samples taken from the patients before and after 3- or 6-month HAART.

RESULTSThe activation percents of CD4(+), CD8(+) T lymphocytes were significantly higher before therapy than the normal controls (HLA-DR/CD4: 40.47 +/- 18.85 vs 11.54 +/- 4.10; CD38/CD4: 81.34 +/- 10.86 vs 53.34 +/- 11.44; HLA-DR/CD8: 63.94 +/- 12.71 vs 25.67 +/- 9.18; CD38/CD8: 86.56 +/- 11.41 vs 58.84 +/- 6.16, all P < 0.01). After 6-month combined antiretroviral treatment, the activation of T lymphocytes in HIV/AIDS patients was significantly decreased (HLA-DR/CD4: 28.31 +/- 13.48; CD38/CD4: 69.88 +/- 12.64; HLA-DR/CD8: 46.56 +/- 18.64; CD38/CD8: 70.17 +/- 14.54, all P < 0.01 compared with the pre-treatment values). Before the treatment, CCR5 expression on CD8(+) T lymphocytes was up-regulated while CXCR4 expression on CD8(+) T lymphocytes downregulated in HIV/AIDS patients compared with the normal controls (CD8/CCR5: 70.91 +/- 10.03 vs 52.70 +/- 7.68; CD8/CXCR4: 24.14 +/- 11.08 vs 50.05 +/- 11.68, all P < 0.01). After 6-month HAART, CCR5 expression on CD8(+) T lymphocytes significantly decreased (56.35 +/- 12.96, P < 0.01), while CXCR4 expression on CD8(+) T lymphocytes increased (36.95 +/- 9.96, P < 0.05) compared with the pre-treatment and the normal controls. A significant statistical relationship was observed between the expression of activation markers, CCR5 and the CD4(+) T lymphocyte counts after HAART (P < 0.05).

CONCLUSIONSReduced activation of T lymphocytes and a normalization of coreceptor expression were observed in Chinese HIV/AIDS patients after HAART. Immunity can be restored in HIV/AIDS patients receiving HAART.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; China ; Female ; HIV Infections ; drug therapy ; immunology ; Humans ; Lymphocyte Activation ; drug effects ; physiology ; Male ; Middle Aged ; Receptors, Chemokine ; analysis ; drug effects ; T-Lymphocytes ; immunology

OBJECTIVETo investigate the safety and efficacy of fast track surgery (FTS) management in gastric cancer undergoing D2 gastrectomy.

METHODSEighty gastric cancer patients undergoing D2 gastrectomy were recruited prospectively. Patients were assigned to receive FTS management (n = 40) or conventional perioperative care (n = 40). The FTS care included shorten preoperative fasting time, no nasogastric decompressing tubes and abdominal drainage placed, early postoperative oral feeding, multimodal analgesia, and early mobilisation. The length of postoperative hospital stay, medical cost, nutritional status, gut function, and postoperative complications in the two groups were recorded and compared.

RESULTSFTS group was associated with a significantly shorter postoperative hospital stay compared with conventional care group [(5.6 +/- 1.3) d vs. (9.4 +/- 1.9) d, P < 0.05]. Medical cost was less [(18 620 +/- 2360) Yuan vs. (20 370 +/- 2440) Yuan, P < 0.05] and duration of intravenous infusion [(3.5 +/- 1.4) d vs. (5.8 +/- 1.9) d, P < 0.05] was also shorter. First passage of flatus was earlier in FTS group than in conventional care group [(4.3 +/- 0.4) d vs. (5.5 +/- 0.9) d, P < 0.05]. Loss of body weight in the postoperative period was less in FTS group [(3.2 +/- 0.8) kg vs. (4.3 +/- 1.6) kg, P < 0.05]. There was no difference in morbidity or mortality between the two groups.

CONCLUSIONFTS in D2 gastrectomy is safe and efficient, and it can shorten postoperative hospital stay and hasten return of gut function.

Adult ; Aged ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Length of Stay ; Male ; Middle Aged ; Perioperative Care ; Postoperative Complications ; prevention & control ; Prospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome

Objective To establish a method for direct orthotopic transplantation of human brain tumors into mouse brain and assess the parental characteristics of the xenografts. Methods Fresh tissues of human brain metastasis of lung adenocarcinoma and glioblastoma tissues subcutaneously implanted in nude mice were harvested and minced into pieces. A special trochar was used to inject the tissue pieces into the right caudate nuclei of nude mice through the burr hole in the mouse skull. The efficiency of tumorigenicity, survival time of tumor-bearing mice, histopathological features, expressions of tumor markers in the xenografts and MRI findings were observed. Results The metastatic lung adenocarcinoma and glioma xenografts were passaged for 6 and 13 generations in nude mice, and the average survival time of the tumor-bearing mice was 38.0±0.9 days and 19.0±1.3 days, respectively. The metastatic xenogratts were characterized by expressions of CEA and acid mucin without invasiveness to the normal recipient brain tissue, which were consistent with the features of the parental metastatic tumor. The glioma xenogratts exhibited deep invasiveness to the normal recipient brain tissue and high expression of EGFR, which were identical to the characteristics of the parental tumor. Conclusion Compared with stereotactic cell suspension injection, direct tumor tissue graft injection requires simple procedures and well maintains the characteristics of the parental tumor tissue in the xenograft in nude mice. This method well suits the purposes of establishing orthotopic xenotransplantation models of brain tumors in nude mice.

This study was designed to evaluate the anti-inflammatory effect of recombinant human kallistatin (Kal) on ulcerative colitis (UC) in the mouse model. Acute colitis was induced by administration of 4% dextran sodium suffate (DSS) to KM mice for 7 days. The mice were then randomized into 5 groups:model control, Kal 0.2 mg·kg^-1·d^-1, 1.0 mg·kg^-1·d^-1 and 2.0 mg·kg^-1·d^-1 group, salazosulfapyridine (SASP) group. Ten age-matched normal KM mouse were administered with saline in the normal control. The weight, colon length, inflammation factor (MPO/SOD/MDA) and TNF-α/IL-10 levels among the five groups of mice were determined. The results showed that histological index score and MPO/MDA/TNF-α levels of high-dose Kal treatment group and SASP group were significantly lower compared with the model group (P<0.01), but the weight, colon length, IL-10 level and SOD activity were significant higher than the model group (P<0.01), approaching the normal group. These parameters showed that Kal can significantly relieve the UC state in a dose-dependent manner. This study demonstrates that Kal significantly remits UC in mice, and participates in the regulation of inflammatory cytokines TNF-α/IL-10 levels and has some antioxidant activity.

Objective: To explore the changes of γ-GCS mRNA expression and GSH-PX in serum of workers exposed to manganese in order to provide scientific basis for early diagnosis of manganese poisoning. Methods: In June 2017, a total of 180 workers from a motorcycle manufacturer were selected by stratified random sampling, including 115 welders as the exposure group and 65 administrative office workers as the Control Group, the exposure group was divided into high exposure group (43 persons) and low exposure group (72 persons) according to whether the exposure group exceeded the standard limit. The levels of γ-gcs Mrna expression and GSH-Px activity in serum were determined by Occupational Health Survey, and the differences of γ-gcs Mrna expression and GSH-Px activity among different groups were analyzed. Results: Compared with the control group, the serum GSH-Px activity was lower and the serum γ-GCS mRNA expression level was higher in the exposed group (F=370.52, 275.95, P<0.01) . Compared with the control group, there was significant difference in γ-GCS mRNA expression level and GSH-Px activity (F=0.475、1.06, P<0.01; F=48.53、111.70, P<0.01) . The concentrations of manganese in air, welding dust and urine were positively correlated with the level of γ-GCS mRNA (r=0.71, 0.50, 0.31, P<0.01) The serum GSH-Px activity was negatively correlated with the concentrations of manganese in air, welding dust and urine (r=-0.80, -0.52, -0.30, P< 0.01) , There was no correlation between Serum γ-GSH-Px activity and age and years of exposure (P>0.05) . Conclusion: Serum γ-GCS mRNA expression level and GSH-Px activity level can be used as early biomarkers of manganese poisoning. The concentrations of manganese in workplace air, welding dust and urine manganese in workers are the influencing factors.
Air Pollutants, Occupational ; Dust ; Humans ; Ions ; Manganese ; Manganese Poisoning ; Occupational Exposure/analysis* ; RNA, Messenger/genetics* ; Welding

Objective Gastrointestinal fistula is a serious complication after operation. It is reported that the over-the-scope-clip(OTSC) can close intestinal full-thickness wall and treat gastrointestinal fistula effectively. This study was to investigate the efficacy of OTSC in the treatment of gastrointestinal fistula.Methods We analyzed the clinical data of 28 postoperative patients with gastrointestinal fistula who underwent endoscopic closure using OTSC in Endoscopy Center of Research Institute of General Surgery in our hospital from October 2014 to October 2017, and recorded the course of disease, the site of gastrointestinal fistula, the diameter of fistula and the efficacy of OTSC. Patients with successful closure were followed up for half a year after hospital discharge to observe the recurrence of fistula.Results Endoscopic closure with OTSC was successful in all the 28 patients without complications like bleeding and falling off. In the sinus tract or digestive tract radiography at 1-2 weeks after OTSC closure, 23 patients were successful in endoscopic closure(82.14%). In the six months′ follow-up of 23 patients with successful treatment, 4 patients were lost to follow-up and 19 patients were successful without recurrence of fistula.Conclusion The endoscopic closure of gastrointestinal fistula using OTSC is safe and effective, avoiding reoperation and reducing trauma.

WRKY, a class of conserved transcription factors in plants, plays important roles in plant growth, development and secondary metabolism. In the present study, 65 WRKY members were identified from de novo transcriptome sequencing data of three different tissues (root, stems and leaves) of Baphicacanthus cusia. BcWRKY proteins contained from 221 to 706 amino acids and the isoelectric point is from 4.68 to 9.68. Molecular weights range from 25 711.8 to 75 475 Da. The main secondary structures of BcWRKYs protein are random coil. A subcellular localization prediction indicated that the putative BcWRKY proteins were enriched in the nuclear region. Phylogenetic analysis showed that BcWRKYs could be categorized into three groups and five subgroups (Group IIa, Group IIb, Group IIc, Group IId and Group IIe) in Group II. Structural analysis found that all BcWRKY proteins contained a highly conserved motif WRKYGQK. Finally, the transcriptional profiles of ten BcWRKY genes highly expressed in root, stem and leaf tissues under abscisic acid (ABA), methyl jasmonate (MeJA), or salicylic acid (SA) treatment were systematically investigated using qRT-PCR analysis. Results showed that a total of ten BcWRKY genes were differentially expressed in response to ABA, MeJA, and SA treatment. This work would be provided a basis for further elucidating the molecular mechanism of WRKY transcription factors in the biosynthesis of indole alkaloids in B. cusia.