Objective To study the preparation and the in vitro and in vivo release profile of GH-Chitosan-Alginate microcapsules.Methods GH-Chitosan-Alginate microcapsules were prepared through impulsive electrostatic technique.The interrelated factors influencing the diameter and sphericity were studied through orthogonal experiments,and finally the statistic analysis made sure the optimum conditions to prepare microspheres.The morphology and size of the microcapsules were observed,and the content,encapsulation efficiency and recovery efficiency of the microcapsules were measured.Moreover,their in vitro and in vivo release experiments were carried out.Results The results showed that the diameter of needle was the most significant factor to the diameter of microspheres.The optimum conditions for the least diameter of microspheres were 450?m diameter of needle,2cm from needle tips to the gelation surfaee,1.5% alginate concentration,8ml/h speed of flowing-liquid and metal containers.The microcapsules had good sphericity morphology and distribution.The size of the microcapsules was in the range of 10-25?m with an average size of 47.93?m.The encapsulation efficiency and GH-load of the microcapsules were 94% and 11.24% respectively.The release kinetics of microcapsules was studied in false gastric and intestines juice.In false gastric juice,the GH of microcapsules was not released;in false intestines juice,it was released well,and TAM was completely released after about 12h.in vivo release profile made sure that the serum GH level of GH microcapsule group was at the highest value(98.59ng/ml) at 8h.The release profile was fitted well in both in vitro and in vivo conditions.Conclusion GH-Chitosan-Alginate Microcapsules have good morphology and sustained release effect.

Objective To investigate the changes of hepatocyte growth factor (HGF) and vascular endothelial growth factor(VEGF) after partial hepatectomy in patients with liver cirrhosis and their relationship with liver regeneration. Methods Thirty-five patients with partial hepatectomy between June 2007 and August 2009 were enrolled,according to whether cirrhosis were divided into cirrhosis group (16 cases) and non-cirrhosis group (19 cases). Liver size were measured with angiographic computed tomography at 7,30,90 d after operation. Regeneration rate of remnant liver were calculated. The serum concentrations of HGF and VEGF were meaaured. Postoperative hepatic function and complications incidence rate were comparatively analyzed. Results Compared with non-cirrhosis group, the postoperative hepatic function of cirrhosis group suffered serious damage. In non-cirrhosis group, the remnant liver regeneration rate reached (63.6± 15.9)%, (79.4 ± 17.2)%, (97.2 ± 18.3)% at 7,30,90 d after operation,in cirrhosis group,it reached (41.7 ± 10.7)%, (55.7 ± 13.2)%, (76.6 ± 12.5)%, liver in non-cirrhosis group regenerated rapidly (P <0.05). After hepatectomy,the HGF levels in cirrhosis group increased significantly at 1,3,7 d than those in non-cirrhosis group(P ＜ 0.05), but the VEGF levels were lower. Conclusions Liver in the patients with cirrhosis regenerate slowly and it may be due to in part through a decrease in VEGF. Whether it may,when given therapeutically represent a strategy to optimize liver regeneration in problematic patients needs to be clarified.

AIM:To establish the quality standard for Xuesaitong Granule(total saponins of Radix et Rhizoma Notoginseng). METHODS:The contents of notoginsenoside R1,ginsenoside Rg1 and Rb1 were determined on the Shim-Pack C 18 (250 mm?4.6 mm,5 ?m) column,with CH3CN—H2O gradual elution and monitored at 203 nm. RESULTS:The average recovery of notoginsenoside R1 was 100.0% (RSD=1.00%,n=6),the minimum detection quantity cannot be lower than 5% labelled weight. The average recovery of ginsenoside Rg1 was 99.8 % (RSD=0.47%,n=6),the minimum detection quantity cannot be lower than 20% labelled weight. The average recovery of ginsenoside Rb1 was 99.8% (RSD=0.79%,n=6),the minimum detection quantity cannot be lower than 30% labelled weight. CONCLUSION:The method is simple,reliable,accurate and can be applied to the quality control of Xuesaitong Granules.

Objective To explore the effect of hypoxia inducible factor-1α silencing by siRNA on proliferation, apoptosis and necrosis of human renal proximal epithelial cell ( HK-2 )under hypoxia. Methods CoCl2 was used to mimic hypoxia, and siRNA technique was used to silence HIF-1α. HK-2 cells were divided into five groups, including normal culture group,hypoxia culture group, transfection reagent group, negative control group and HIF-1α siRNA group.MTT assay was used to detect the growth inhibition ratio of cells. TUNEL and caspase-3 quantity was used to detect apoptosis. LDH activity in supernatant was detected to evaluate cell necrosis.Real-time PCR and Western blotting were used to detect mRNA and protein of HIF-1α, HIF-2α,glucose transporter 1(Glut-1) and vascular endothelial growth factor (VEGF). Results (1) The transfection efficiency of siRNA was 95%-100%. Under normoxia, the efficiency of siRNA silencing HIF-1α gene reached to 70%, while under hypoxia, it was 97%. (2) The growth inhibition ratio of cells in HIF-1α siRNA group was significantly higher than that of other groups including hypoxia culture group, transfection reagent group and negative control group (all P＜0.05). No significant difference was found in apoptotic ratio of the other four groups except normal culture group (P＞0.05). The LDH level in HIF-1α siRNA group was significantly higher than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). (3) The expression of HIF1α, Glut-1, VEGF mRNA and protein in HIF-1α siRNA group was significantly lower than that of hypoxia culture group, transfection reagent group and negative control group (P＜0.05). No significant difference was observed on the level of HIF-2α mRNA and protein in the other four groups except normal culture group (P＞0.05). Conclusion HIF-1α gene silencing can inhibit the mRNA and protein expression of Glut-1 and VEGF, and can aggravate growth inhibition and necrosis of HK-2 cells under hypoxia.

Objective To investigate the location and expression of hypoxia inducible factor (HIF) subunits in the remnant kidney of 5/6 nephrectomy rats. Methods Remnant kidneys were produced in adult male SD rats by 5/6 nephrectomy. The renal function and histopathological changes were evaluated at week 1, 2, 4, 6, 8 and 12 after operation. Tissues of remnant kidneys were collected to detect the location and expression of HIF-1α and HIF-2α by immunohistochemistry staining and Western blotting. The mRNA levels of HIF targeted genes vascular endothelial growth factor (VEGF) and heme oxygenase-1 (HO-1) were determined by RTPCR. Results (1) 5/6 nephrectomy rats underwent one week of acute renal failure at first[Scr (122.8±22.1) μmol/L] and then developed compensative chronic renal failure [(66.0±3.7)-(66.4±8.4) μmol/L], but the level of Scr increased quickly after week 6 [(66.4±8.4)-(127.8±22.7) μmol/L],concomitantly with progressive tubulointerstitial fibrosis in remnant kidney cortex. (2) In cortex, HIF-1α was expressed only in the atrophic and dilated tubular cells while HIF-2α was located in endothelial, interstitial fibroblasts, and vascular smooth muscle cells. The semiquantitative results of imunohistochemistry and Western blotting revealed that HIF-1α and HIF-2α were both gradually up-regulated during the early stage of remnant kidney, peaked at week 4 and 6, and then gradually down-regulated. (3) The mRNA levels of HIF targeted genes VEGF and HO-1 transiently peeked at week 4 and 6, and then decreased gradually. Conclusions The increased stabilization of HIF-αprotein and transcription of HIF targeted genes at the early stage of this model is a compensation reaction towards hypoxia. The mechanism of decreased expression of HIF-α at the end stage of chronic kidney disease deserves further investigation.

AIM: To investigate the effects of IL-13 on expression of IL-1? in acute renal ischemia/reperfusion injury. METHODS: Fifty-seven male Wistar rats were randomly divided into 8 group: normal group,sham operation group, ischemia group, ischemia/reperfusion injury group(I/R), normal saline(NS)-treated group 1(C-1), NS-treated group 2(C-2),IL-13-treated group1(T-1)and IL-13-treated group 2(T-2).Rats were subjected to 45 min bilateral renal ischemia followed by reperfusion. rmIL-13 (1.5 ?g/50 g body weight )was injected into the renal arteries through the abdominal aorta before ischemia(T-1) or immediately afterischemia(T-2).The serum level of IL-1? and the renal expression of IL-1? were determined in each group at 24 h post-ischemia. In addition, BUN,Cr and renal histology were also measured. RESULTS: (1)The serum level of IL-1? [C-1to T-1: (27.13?5.51) ng/L to (14.05?3.82) ng/L, P

Objective To identify the metabolite levels and to study the potential effects of risporidone tablets on prefrontal lobe and thalamus in male first-episode schizophrenia by proton magnetic resonance spectroscopy (1H-MRS).Methods Twenty-two male first-episode schizophrenics were examined at prefrontal lobe and thalamus by multi-voxel 1H-MRS before and after 8 weeks' risperidone tablets treatment,and 30 normal controls were assessed once.The patients were also received positive and negative syndrome scale(PANSS) and Wisconsin card sorting test (WCST) before and after treatment.The N-aeetylaspartate ( NAA ),Choline-congtaining compounds (Cho),and Creatine compounds (Cr) were measured and the ratios of NAA/Cr,Cho/Cr were determined.Results On left prefrontal lobe and left thalamus,the NAA/Cr ratio in patients before treatment demonstrated lower than those in normal controls ( ( 1.37 ± 0.33 ) vs ( 1.61 ± 0.38 ),t =2.57,P ＜ 0.05 ; ( 1.46 ± 0.35 ) vs ( 1.71 ± 0.38 ),t =2.36,P ＜ 0.05 ).There were no significant differences in both the ratios of NAA/Cr and Cho/Cr between pre- and post-treatment (P＞0.05 ).The changes of NAA/Cr on left prefrontal lobe post-treatment were negatively related with the alterations of the total score of PANSS ( r =- 0.46,P ＜ 0.05 ),the score of negative symptoms ( r =- 0.48,P ＜ 0.05 ),the responses errors ( r =- 0.42,P ＜ 0.05 ) and the porseverative errors ( r =- 0.40,P ＜ 0.05 ),meanwhile,positively related with the alterations of the categories completed ( r =0.44,P ＜0.05 ) and the conceptual level responses ( r =0.42,P ＜ 0.05 ).Conclusions Abnormalities in neuronal function and/or integrity presented on prefrontal lobe and thalamus maybe exist in male first-episode schizophrenics.Short-term antipsvchotic treatment with risperidone tablets may have no effects on measures of prefrontal lobe and thalamus.

Objective To compare the metabolic measures on prefrontal lobe and thalamus among schizophrenics with or without mental disorder family history by proton magnetic resonance spectroscopy (1 H-MRS),and to explore the relationship among metabolic measures,clinical symptom,and executive functioning.Methods Thirty-one schizophrenics with schizophrenia family history,21 schizophrenics with the other mental disorder family history,and 78 schizophrenics without mental disorder family history were examined at prefrontal lobe and thalamus by multi-voxel 1H-MRS.The N-acetylaspartate (NAA),Choline-congtaining compounds (Cho),and Creatine compounds (Cr) were measured and the ratios of NAA/Cr and Cho/Cr were determined.Meanwhile,Positive and Negative Syndrome Scale (PANSS) and Wisconsin Card Sorting Test (WCST) were also assessed in all schizophrenics.Results Both in schizophrenics with schizophrenia family history and with the other mental disorder family history,the NAA/Cr ratios showed lower than those in schizophrenics without mental disorder family history both on left prefrontal lobe and on fight thalamus ( P ＜ 0.05 ).Compared with schizophrenics without mental disorder family history,the score of negative symptoms and the perseverative errors demonstrated higher ( P＜ 0.05 or 0.01 ),the categories completed showed lower both in schizophrenics with schizophrenia family history and with the other mental disorder family history ( P＜0.05 ).The NAA/Cr ratios on left prefrontal lobe in all schizophrenics were significantly negatively related with the total score of PANSS and the responses errors (P ＜ 0.05 or P＜ 0.01 ),and positively related with the categories completed and the conceptual level responses ( P＜ 0.05 or P＜ 0.01 ).On left prefrontal lobe both in schizophrenics with schizophrenia family history and with the other mental disorder family history,the ratios of NAA/Cr were negatively related with the score of negative symptoms and the perseverative errors ( P ＜ 0.05 or 0.01 ).Conclusion The damages of neurons on prefrontal lobe and thalamus in schizophrenics with mental disorder family history may be more severe than those in schizophrenics without family history,and the damages on prefrontal lobe are related with negative symptoms and executive functioning.

Objective To compare the differences between the schizophrenics with negative and positive symptoms on prefrontal lobe and thalamus by proton magnetic resonance spectroscopy ( 1 H-MRS). Methods 58 negative subtype and 51 positive subtype schizophrenics were examined at prefrontal lobe and thalamus by multivoxel 1H-MRS before antipsychotic treatment The N-acetylaspartate (NAA), choline-containing compounds ( Cho), and creatine compounds (Cr) were measured and the ratios of NAA/Cr, Cho/Cr were determined. Results On right thalamus,the NAA/Cr ratio in negative subtype patients ( 1.40 ± 0.29 ) demonstrated lower than that in positive subtype ( 1.62 ± 0.33 ), the same phenomenon were appeared on male, female, non-first-episode, with-medicine and without-medicine patients (P ＜ 0.05 or 0.01 ). The Cho/Cr ratio on right thalamus in negative subtype of female,non-first-episode,without-medicine schizophrenics were lower than those in positive subtype (P ＜ 0.05 ).On left prefrontal lobe,left thalamus and right thalamus, the NAA/Cr ratios both in negative subtype and positive subtype schizophrenics were significantly negatively related with age of onset(P＜0. 05 or 0.01 ). In negative subtype schizophrenics,the Cho/Cr ratio on right thalamus was positively related with age of onset ( r = 0. 25, P ＜0.05 ). In negative subtype schizophrenics of non-first-episode, without-medicine, the correlation was negative between the NAA/Cr ratio and the course of disease( r= -0.48, -0.46, P＜0.05 ) ,and was positive between Cho/Cr ratio and the course of disease on right thalamus( r= 0.58,0.56, P＜ 0.01 ). Conlusion Compared with positive subtype schizophrenics,negative subtype schizophrenics have greater impairments on 1 H-MRS on right thalamus. The course of disease has greater effects on 1 H-MRS in negative subtype schizophrenics.

Objective To construct a prokaryotic plasmid to express the testis development related gene 1 (TDRG1) recombinant protein and obtain anti-TDRG1 mAb by immunizing mice, and to identify the biological properties of the mAb. Methods The coding sequence of TDRG1 was amplified by RT-PCR from normal human testis tissue and cloned into the vector pET21, and then was expressed in the E. coli BL 21(DE3) to get TDRG1 recombinant protein. The purified TDRG1 recombinant protein was injected to immunize the BALB/C mice to develop anti-TDRG1 mAb. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted anti-TDRG1 mAb were subcloned with limited dilution. Enzyme-linked immunosorbent assay (ELISA) was used to evaluate titers and subtypes of mAb. Western blot and immunohistochemistry were used to detect specificity of mAb.Results The prokaryotic plasmid expressing the recombinant protein was constructed, and the TDRG1 recombinant protein was expressed and purified. Two hybridoma cell lines that secreted anti-TDRG1 mAb were obtained. The titer of the mAb in ascites was 1∶1.6×10~6, and the subtype of the mAb was IgG_1. Westem blot and immunohistochemistry analysis indicated the mAb showed specific combination with TDRG1 protein in human testes.Conclusion The TDRG1 recombinant protein is highly purified and has strong antigenicity. The anti-TDRG1 mAb is produced successfully.