Objective To reduce experimental costs and improve the utilization of S9, we use spiral coating technique to evaluate the activity of rat liver S9 prepared by combination inducing method as well as to establish cryopreservation method.Methods Using spiral coating technique and Ames test to evaluate the activity of self-made rat liver S9 and commercially available S9 separately.We use glycerinum as protective agent to establish cryogenic storage method, so that S9 can be in liquid form stored at -20 °C in the refrigerator.Results In the Ames assay as well as using spiral coating technique, the number of revertant colonies had dose-response relationship among the dose of S9.When conditions were the same, the number of revertant colonies in positive control was at the approximate level in presense of self-made rat liver S9 and commercially available S9 respectively.When S9 ( concentration of 38%) was added to the amount at 1.48 ~6.62 μL /μL broth dose of bacteria, it can significantly induced Salmonella typhimurium histidine strains TA100, reverse mutation rates were three times more than the control group.Conclusions Spiral coating technique can successfully evaluate the activity of rat liver S9.The inducing method of combination of PB and BF can take the place of the unducing method of PCBs in the preparation of Liver homogenate S9.

Adult ; Female ; Humans ; Lacrimal Apparatus ; immunology ; Lymphocytes ; immunology ; Male ; Mucous Membrane ; immunology

The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

BACKGROUND: Direct anterior minimum invasive surgery (DAMIS) total hip arthroplasty has been considered as the total hip arthroplasty that is most consistent with the minimally invasive standards. In theory, there is a low incidence of complications in the DAMIS total hip arthroplasty. However, because minimally invasive surgery demands high surgery skills, the incidence rate of complications is high in the early period of its learning curve. Lateral decubitus position DAMIS was considered to provide a better perspective, so that, the incidence of surgical complications may be reduced. OBJECTIVE: To explore the feasibility of lateral decubitus position DAMIS for total hip arthroplasty. METHODS: Totally 62 cases from the Department of Orthopedics, First Affiliated Hospital of Gannan Medical University between April 2013 and April 2016 were retrospectively analyzed, and randomly assigned to two groups: lateral decubitus position group (n=31) and supine position group (n=31). Incision length, operative time, intraoperative blood loss, acetabular cup angle, hospital stay, Harris scores, Visual Analogue Scale score, and SF-36 score were compared between the two groups. Complications at postoperative 1 year were recorded. RESULTS AND CONCLUSION: (1) Operative time was longer and intraoperative blood loss was more in the lateral decubitus position group than in the supine position group (P < 0.05). There were no statistically significant differences in incision length and hospital stay between the two groups (P > 0.05). (2) There were no significant differences in the incidence of postoperative complications between the two groups (P >0.05). (3) There were no significant differences in the anteversion and abduction angles of acetabular cup between the two groups (P > 0.05). (4) There were no significant differences in Harris hip score, Visual Analogue Scale score, and SF-36 the quality of life scores between the two groups before surgery and 1 year after surgery (P > 0.05). (5) In conclusion, DAMIS total hip arthroplasty can abandon special traction bed when patients are in lateral decubitus position. The short-term effects are the same with supine DAMIS total hip arthroplasty.

BACKGROUND:There are few reports concerning effects of warming the yang and benefiting the marrow for the knee osteoarthritis on the expression of matrix metal oproteinase.
OBJECTIVE:To observe the effect of warming the yang and benefiting the marrow on the expression of matrix metal oproteinase in rabbit models of knee osteoarthritis.
METHODS:Of 96 healthy adult New Zealand rabbits, 72 rabbits were randomly selected for making rabbit models of knee osteoarthritis using plaster external fixation. After success model establishment, the rabbits were randomly assigned to three groups. Model group was left intact. Chinese medicine group received daily intragastric administration of drug extract 24 mL/kg. Drug control group was daily intragastrical y administered Puli Capsule (glucosamine hydrochloride) 24 mg/kg, once a day, until the eighth week of success model induction. An additional 24 New Zealand rabbits served as blank controls.
RESULTS AND CONCLUSION:Using quantitative PCR, matrix metal oproteinase-1, matrix metal oproteinase-3 and matrix metal oproteinase-13 expression was significantly higher in the model group than that in the other three groups. Matrix metal oproteinase-1, matrix metal oproteinase-3 and matrix metal oproteinase-13 expression was significantly lower in the Chinese medicine group and drug control group than that in the model group. These results indicated that warming the yang and benefiting the marrow for knee osteoarthritis in rabbits could effectively inhibit the expression of matrix metal oproteinase in rabbits.

Objective The purpose of this study was to prepare RGD4C modified ferritin nanocages (RGD4C-FRT) for targeted drug delivery to rat hepatic stellate cells (HSC-T6). Methods RGD4C modified human H-chain ferritin was expressed and purified. Doxorubicin (Dox) was encapsulated into the cavity of RGD4C-FRT through ion channels, which resulted in RGD4C-FRT-Dox. The target of RGD4C-FRT-Dox to HSC-T6 was detected using fluorescence microscopy. Results Transmission electron microscopy showed that RGD4C-FRT was hollow spherical-structured with uniform size and good dispersion. The average particle diameters of RGD4C-FRT and RGD4C-FRT-Dox were 12.57 nm and 20.13 nm , respectively. The drug encapsulation efficiency and loading percentage of RGD4C-FRT-Dox were 77.32% and 15.88% respectively. RGD4C-FRT-Dox was significantly uptaken by HSC-T6, and the uptake could be blocked by the empty carrier RGD4C-FRT. Conclusion RGD4C-FRT-Dox can specifically target HSC-T6. Further animal experiments are needed to inspect its therapeutic effect on liver fibrosis.

s:The chemical constituents of gliocladium roseum(called Y)accelerating the growth of famous medicin al plant Anoectochilus roxburghiiwas studied.Five comp ounds were separated by silica gel column chromatograph from this fungal mycelia and their structures were elu cidated by the data of IR,NMR,UV and MS.Compound I was 6,22-diene-3-hydroxy- 5,8-epidioxy ergosta,compound 2 is ergosterol,compound 3 is D-arabitol and com pound 4 is mannitol.

To investigate the effects of albumin on the production of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) in podocytes. Podocytes were treated with bovine serum albumin (BSA) at the concentration of 0.1, 0.5, 1, 2 g/L, respectively. Conditioned media were harvested 12, 24, 48 and 72 h after the treatment. The expression of MMP-2 and MMP-9 was assayed by gelatin zymography, RT-PCR and Western blotting analysis. Our results showed that in comparison with the control group, BSA increased the expression of MMP-2 and MMP-9 mRNA and protein in a dose- and time-dependent manner (P<0.05). Meanwhile, the enzymatic activities of MMP-2 and MMP-9 in the culture supernatants of podocytes were also increased (P<0.05). It is concluded that albumin up-regulated the expression of MMP-2 and MMP-9 at gene and protein levels in a time- and dose-dependent manner.

This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy (AN). A total of 36 healthy male SD rats were randomly assigned to three groups: control group, AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin (6.5 mg/kg) in both AN group and sulodexide treatment group. Sulodexide (10 mg/kg) was administered the rats in the treatment group once daily by garage from the first day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14, 28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin, CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and protein were increased in the glomeruli of AN rats at day 14 and 28 after the model establishment, which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And, the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide- treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP (as well as 24-h urinary protein excretion). It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.

Objective To access the effects of aldosterone (ALD) and its receptor antagonist- spironolactone (SPI) on the production of reactive oxygen species (ROS) and apoptosis in podocytes and to explore the possible mechanism involved. Methods Conditionally immortalized mouse podocytes were divided into control group, ALD group, SPI group and SPI +ALD group. The level of ROS production and the expression of nephrin protein were assayed by fluorescent spectrophotometry and indirect immunofluorescence technology. The apoptosis rate of podocytes was monitored by flow cytometry. The expression of bax and bcl-2 mRNA and protein was detected by RT-PCR and Western blotting methods. The anti-oxidant N-acetylcysteine (NAC)was applied to determine whether the effects of ALD on podocytes were mediated by ROS pathway.Results Compared with the control group, ALD significantly increased ROS production in podocytes (P＜0.05). SPI completely abolished the above-mentioned effect of ALD (P＜0.05). ALD induced the down-regulation of the expression of nephrin and the up-regulation of podocytes apoptosis (P＜0.05), which was accompanied with the elevated expression of bax mRNA and protein and the reduced expression of bcl-2 mRNA and protein (P＜0.05). SPI or NAC prevented the above-mentioned changes induced by ALD (P＜0.05). Conclusion ALD increases theexpression of pro-apoptotic factor (bax) but decreases the expression of anti-apoptotic factor (bcl-2)to induce podocytes apoptosis through the mineralocorticoid receptor (MR) possibly via the mechanisms involving ROS pathway.