Objective To observe the effect of stageⅢdiabetic nephropathy(DN)treated by Qizhi Jiangtang capsule and explore its potential mechanism. Methods According to digital table method,the patients who conformed to the diagnostic criteria of stageⅢDN were randomly divided into two groups:an experiment group and a control group. All the patients in the two groups took elution treatment for 2 weeks,and then were treated with western basic therapy. The patients in the experiment group were administered orally with Qizhi Jiangtang capsule(2.5 g once, 3 times a day),while those in the control group treated with valsartan 80 mg,once a day. Urine microalbumin(mALB), mALB/urine creatinine(UCr),β2-microglobulin(β2-MG),α1-microglobulin(α1-MG)were observed in the two groups,endothelin-1(ET-1),nitric oxide(NO),thromboxane B2(TXB2),6-keto prostaglandin F1α(6-keto-PGF1α) were also determined. Serum creatinine(SCr),blood urea nitrogen(BUN),serum cystatin-C(Cys-C),retinol-binding protein(RBP),β2-MG were detected in the blood biochemistry automatic analyzer. These laboratory markers were inspected before treatment and at the 4th,8th and 12th week after treatment. Results Ninety-six patients in the experiment group and 95 patients in the control group were effectively included in the end. Before treatment,there were no statistic significant differences in urine mALB,mALB/UCr,β2-MG,α1-MG and blood ET-1,NO,TXB2, 6-keto-PGF1α between two groups(all P>0.05). Along with the prolongation of treatment,urine mALB,mALB/UCr,β2-MG,α1-MG and ET-1,TXB2 were significantly reduced,while NO,6-keto-PGF1α were significantly raised in the two groups after treatment,and the above changes in the experimental group were more obvious. There were statistic significant differences of mALB,mALB/UCr,β2-MG,α1-MG and TXB2,6-keto-PGF1αbetween two groups at the 12th week after treatment〔mALB(mg/L):36.6±9.2 vs. 78.6±16.5,mALB/UCr(mg/mmol):3.90±1.97 vs. 9.70±2.90,β2-MG(mg/L):0.25±0.10 vs. 0.40±0.12,α1-MG(mg/L):8.40±2.26 vs. 12.50±3.21,TXB2 (ng/L):75.8±18.7 vs. 94.7±21.7,6-keto-PGF1α(ng/L):73.4±15.2 vs. 65.2±11.5,P<0.05 or P<0.01〕. But there were no statistic significant differences of ET-1 and NO between experimental group and control group at the same time-points〔ET-1(ng/L):57.6±6.9 vs. 59.1±6.2,NO(μmol/L):68.9±11.6 vs. 65.4±10.7,both P>0.05〕. In each of the two groups,the comparisons of the levels of SCr,BUN before and after treatment,there was no statistical significant difference at any time point;the same comparisons between the two groups,there was also no statistic significant difference before treatment and at each of the same time-point after treatment(all P>0.05). The levels of Cys-C,RBP andβ2-MG of the control group after treatment had the tendency of decreasing,but no statistic significant differences were found(all P>0.05). The levels of Cys-C,RBP,β2-MG of the experimental group at the 12th week after treatment were significantly lower than those before treatment〔Cys-C(mg/L):0.72±0.07 vs. 0.89±0.12,RBP (mg/L):53.0±14.2 vs. 66.1±16.5,β2-MG(mg/L):1.86±0.71 vs. 2.79±0.82,all P<0.05〕. Conclusions Qizhi Jiangtang capsule can significantly reduce the levels of urine mALB and mALB/UCr of patients with stageⅢDN and stabilize their renal functions;its therapeutic effect is better then that of valsartan. Its mechanisms are related to the reduction of ET-1,elevation of NO,maintenance of dynamic equilibrium of thromboxane A2/prostacycline(TXA2/PGI2) and protection of vascular endothelial cells.

Objective To construct a plasmid DNA encoding G glycoprotein of respiratory syncytial virus(RSV) and investigate the protective immune response against RSV infection. Methods Recombinant plasmid DNA of pcDNA3.1~G was constructed by standard RT-PCR based cloning procedure. The immunogenicity of recombinant G protein transiently expressed in HEK293 cells was detected by Western blot. BABL/c mice were intramuscularly immunized with pcDNA3.1~G. Samples of lung, sera, bronchoalveolar lavage fluid(BALF) were collected before and after RSV challenge; virus titer in lung was detected by viral titration; sections of paraffin embedding lung tissues were stained by haematoxylin and eosin(HE) for histological analyses; sera anti-RSV IgG levels were examined by ELISA; Th1/Th2 cytokine were detected by ELISA kit, the T lymphocyte subsets of BALF was determined by immunefluorescence staining followed by flow cytometry. Results Plasmid DNA of pcDNA3.1~G was successfully constructed. The expressed target protein possesses immunogenicity. After challenge, pcDNA3.1~G immunized mice presented relieved pathological changes in lung as well as reduced lung viral titers. The RSV specific IgG was detected in sera of immunized mice. There was significantly increased number of CD25~+CD4~+ T cells in mice BALF. Conclusion We constructed a pcDNA3.1~G plasmid DNA vaccination which can induce evident protective cellular immunity against RSV infection in mice with the increased number of CD25~+CD4~+ T cell subpopulation.

Objective To study the prevention and treatment effect of ω-3 fatty acids on intestinal mucosa in critical illness. Methods Forty patients including severe trauma, infection shock were enrolled as experimental group, while 30 healthy people as control group. At the same time, the patients in expermental group were randomly divided into group A and group B(20 cases each). While the patients were. Treated with low calorie parenteral nutrition totally, those in group A received ω-3 fatty acids additionally. The plasma concentrations of dimnine oxidase (DAO), endotoxin were detected by spectrophotography, and TNF-α was detected by ELISA. Results After treatment the concentration of DAO, endotoxin, TNF-α in group A and that of endotoxin in group B decreased significantly (P<0.05 ). While there was no significant difference of endotoxin levels between group A and group B. After therapy, DAO and TNF-α levels in group A were sig-nificantly lower than those in group B (P<0.05 ). The concentrations of DAO and TNF-α in group B were also significantly higher than those in control group (P<0.05). Conclusion ω-3 fatty acids can prevent and treat critical intestinal mucosa effectively.

Objective To investigate the effects of intrathecally coadministered dexamethasone and spironolactone in trathecally on radicular pain behaviors.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion (CCD) ,48 male SD rats successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into Sham-operation group (Sham group, n= 12),Control group ( C group, n = 12 ), Dexamethasone group ( D group, n = 8 ), Spironolactone group ( S group, n = 8 )and Dexamethasone plus spironolactone group (DS group, n=8).Rats in D group,S group or DS group were intrathecally treated with dexamethasone 4 μg, spironolactone 3 μg or dexamethasone 4 μg plus spironolactone 3 μg twice daily on day 2 ～4 after CCD respectively,while rats in C and Sham group received 10μl 10% alcohol.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before CCD and day 1,4,7,10,14,17 and 21 after CCD.Results Compared with Sham group, both PWMT and PWTL were significantly decreased after CCD surgery on the ipsilateral side(P＜0.01 ＝.Intrathecally administrated with dexamethasone significantly improved pain behaviors (P＜0.01 ＝ and these therapeutic effects lasted up to 10 days after CCD surgery.As with dexamethasone,intrathecal spironolactone also significantly attenuated PWMT (P＜0.01 ＝ and PWTL (P＜0.01 ＝ and the change lasted up to 7 days after CCD surgery.Coadministration spironolactone and dexamethasone exhibited significant synergies( PWMT: ( 13.52 ± 0.72) g vs ( 11.58 ± 1.38 ) g, P ＜0.01; PWTL: ( 19.63 ± 1.68) s vs ( 14.14 ± 1.52) s, P ＜ 0.01 ＝.These effects lasted up to at least 10 days.Conclusion Both dexamethasone and spironolactone intrathecally have therapeutic effects on radicular pain behaviors, combination injection of these two drugs could generate significant synergies.

Objective To explore the high-risk factors,clinical characteristics,therapy and prognosis of invasive fungal infection (IFI)in patients underwent allogeneic haemopoietic stem cell transplantation (AlloHSCT). Methods One hundred patients underwent Allo-HSCT at our department from March 2002 to July 2010 were analyzed retrospectively,among whom 26 patients had invasive fungal infection(IFI). Seven patients had pulmonary IFI before allo-HSCT, 14 patients had pulmonary IFI after allo-HSCT,3 patients had respiratory tract system IFI, and 2 patients had intestinal IFI. We observed the occurrence of Graft-versus-host disease (GVHD) ,cytomegalovirus( CMV )infection, Lymphocyte subsets and chronic basic diseases in patients with IFI. The twenty six cases were divided into two groups: experience therapy group with 12 cases and preemption therapy group with 14 cases. Results Among 26 patients with IFI,20 cases suffered from GVHD,6 cases had CMV infection,19 cases had low cellular immune function simultaneously. 1 case had diabetes,3 patients had pulmonary tuberculosis and 1 case had bronchiectasis as complications. In experience therapy groupe: 8 cases (67%)recovered completely but 1 case(8% )suffered from progressive infection. In preemption therapy groupe:3 cases ( 21% ) recovered completely but 5 cases ( 36% ) suffered from progressive infection. Conclusion Clinician should pay close attention to the patients with high-risk factors of IFI after allo-HSCT.

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7, and investigate whether hepatocyte growth factor (HGF) is involved in this process. Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion. On the basis of the co-culture, the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell. The difference in the HGF expression between them was detected by ELISA. The secretion of HGF was knocked down by using RNA interference technology in CAFs. Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell. Eventually, we isolated high-purity CAFs and NFs, and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs. ELISA results demonstrated that CAFs secreted higher HGF, and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference. It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

Objective To investigate the analgesic effects of intraperitoneal lithium chloride injection on radicular pain behaviors in rats.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion(CCD) ,40 male SD rats were randomly divided into model group and Sham-operation group (group S, n= 12) of radicular pain were established.The rats in the model group were subdivided randomly into Control group(group C, n= 12) ,Early treatment group(group E, n=8) and Later treatment group(group L, n= 8 ).Rats in group E were intraperitoneal injected with lithium chloride once daily on day 2 ～ 4 after CCD respectively,while rats in L,group C and S treated with Vehicle(0.9% NaCl).Rats in L group were intraperitoneal treated with lithium chloride on day 12 ～ 14 after CCD respectively,while rats in E,group C and S received Vehicle.The pain ethology indexes such as paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before operation and day 1,4,7,10, 14, 17 and 21 after operation.Results Compared to S group and preoperative level, PWMT and PWTL decreased at Day 1 postoperative in group C (P＜0.05).At day 4 after the operation,compared with group C(7.712 ±0.237)g and (8.190 ±0.382) s,PWMT and PWTL of E group increased to ( 14.607 ± 0,280) g and ( 19.940 ± 0.933 ) s (P ＜ 0.05 ) after intraperitoneal injected lithium chloride.At day 14, compared with group C ( 6.788 ± 0.331 ) g and ( 7.301 ± 0.481 ) s, PWMT and PWTL of group L increased to ( 11.700 ± 0.379) 8 and ( 18.524 ± 1.060) s (P ＜ 0.05 ).This analgesic effect of lithium chloride continued to exist at day 21.However, there was still a significant difference between S group and E,group L(P＜0.05).Conclusion Intraperitoneal lithium chloride injection alleviates pain behavior on radicular pain in rats.

BACKGROUND/OBJECTIVES: D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo. MATERIALS/METHODS: The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis. RESULTS: At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-kappaB, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice. CONCLUSIONS: Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.
Animals ; Apoptosis ; Asia ; Asian Continental Ancestry Group ; Blotting, Western ; Caspase 3 ; Caspase 9 ; Chromatin ; Colon ; Colonic Neoplasms ; Down-Regulation ; Flow Cytometry ; HCT116 Cells ; Humans ; Inflammation ; Methanol ; Mice* ; Neoplasm Metastasis ; NF-kappa B ; Up-Regulation

This study was aimed to explore the influence of breast cancer associated fibroblasts (CAFs) in migration and invasion of breast cancer cell line MCF-7,and investigate whether hepatocyte growth factor (HGF) is involved in this process.Primary breast CAFs and their corresponding normal breast fibroblasts (NFs) were obtained by collagenase digestion.On the basis of the co-culture,the migration and invasion capacity of MCF-7 cells was compared between CAFs and NFs by Transwell.The difference in the HGF expression between them was detected by ELISA.The secretion of HGF was knocked down by using RNA interference technology in CAFs.Then the changes of migration and invasion capacity of MCF-7 cells were investigated by Transwell.Eventually,we isolated high-purity CAFs and NFs,and the CAFs had a stronger ability in promoting MCF-7 migration and invasion than the NFs.ELISA results demonstrated that CAFs secreted higher HGF,and the capacity of MCF-7 migration and invasion was declined after knocking down the secretion of HGF in CAFs by RNA interference.It is suggested that CAFs can promote MCF-7 migration and invasion through HGF in vitro.

Objective To investigate the differences in expression profile of long non-coding RNA (lncRNA) in retina between normal newborn mice and those with oxygen-induced retinopathy (OIR) to lay a foundation for further study of regulatory mechanisms of lncRNA in retinal neovascularization, and to provide a new theoretical basis for the prevention and treatment of retinopathy of prematurity (ROP). Methods A total of 48 cleaning grade C57BL/6J neonatal mice were randomly divided into two groups with 24 in each: OIR group and control group. Those in the OIR group were exposed to hyperoxia [(75±2)％ O2] from 7th to 12th day after birth, and then re-exposed to normoxia (room air) for five days to establish a OIR mouse model. Mice in the control group were raised in room air all the time. Retinal tissues were collected from each mouse on the 17th day after birth. Retinal angiography and hematoxylin-eosin (HE) staining were performed to confirm the establishment of OIR mouse model. Expression profiles of lncRNAs were analyzed by lncRNA high-throughput sequencing and the results were verified by real-time fluorescence quantitative polymerase chain reaction (PCR). Based on the expression of lncRNAs and mRNA and the relationship between their genomic locations, potential target genes of lncRNAs were obtained. Gene ontology analysis (GO) and Kyoto Encyclopedia of Genes and Gnomes (KEGG) pathway analysis were used for bioinformatics analysis of lncRNA expression profile in OIR. T test and Graphpad prism were used for statistical analysis and spreadsheet. Results Retinal angiography and HE staining identified serious retinal damage in the OIR group. In total, 1118 differentially expressed lncRNAs ( ≥ 2.0-fold difference in expression, P<0.05) relating to OIR between the two groups were identified by lncRNA high-throughput sequencing. Results of real-time fluorescence quantitative PCR confirmed that XLOC_150632 and XLOC_150636 were upregulated in the OIR group as compared with those in the control, while XLOC_122045,XLOC_100454, XLOC_170009 and XLOC_122042 were downregulated. GO analysis showed that 852, 1148 and 2100 target genes of differentially expressed lncRNAs were relating to biological process, cellular component and molecular function, respectively. KEGG analysis showed that the target genes were associated with 36 biological pathways, mainly relating to retinal development, chemotaxis, migration and energy metabolism of vascular endothelial cells. Conclusions lncRNAs are differentially expressed in healthy newborn mice and newborn mice with OIR. Fibroblast growth factor 2 (FGF2), the potential target gene of XLOC_150632 and XLOC_150636, is related to phosphatidylinositol-3-kinase (PI3K) signaling pathway and may be involved in retinal neovascularization. This might be a potential target for ROP prevention and treatment.