ObjectiveTo investigate the effects of carboxymethyl-chitosan(CMCS) on chondrocyte apoptosis induced by sodium nitroprusside (SNP),and the effects of CD44 in the process.MethodsA small interference RNA (siRNA) targeting to CD44 mRNA (siRNA-1,siRNA-2,siRNA-3) was constructed.The siRNA was transfected into chondrocytes in vitro with LipofectamineTM 2000.The efficacy of transfection was detected by transfecting fluorescence siRNA into cells.The mRNA expression of CD44 in vitro was detected by RT-PCR.The protein level of CD44 was detected by Western blotting.The apoptosis rate of the transfected and non-transfected cells induced by SNP was detected by flow cytometry.Statistical analysis was conducted with one-way ANOVA and SNK-q test.ResultsThe efficacy of transfection was about 60％.As compared with the control group,the mRNA expression was specifically inhibited after transfecting CD44 siRNA-1 for 24,48 and 72 h(0.198±0.007 vs 0.429±0.053 at 24 h,0.211±0.016 vs 0.501±0.037 at 48 h,0.153±0.005 vs 0.341±0.009 at 72h,q=5.93,7.01,11.23,P＜0.01 ),and the protein level of cells was inhibited after transfecting CD44 siRNA-1 for 24 h compared with the control group (0.231±0.064 vs 0.675±0.113,q=13.09,P＜0.01 ).The FCM results showed that 3 mmol/L SNP could induce chondrocytes apoptosis(70±6)％,and 50,100,200 μg/ml C MCS could affect the inhibitory effect of SNP-induced apoptosis of chondrocyte [ (51 ±7)％,(30±4)％,(15±4)％,q=5.08,6.97,9.73,P＜0.01 ],but it had milder inhibitory effect on CD44-siRNA-1 transfected chondrocytes when compared with those of the non-transfected chondrocytes [ (34±6)％ vs(15±4)％,q=6.95,P＜0.01 ].ConclusionThe data of this study has suggest that siRNA-1 against CD44 gene can significantly inhibit the expression of CD44 in chondrocyte of rats in vitro after transfection.The CD44 may play an important role in chondrocyte apoptosis induced by SNP and protected by CMCS.

Objective To investigate the clinical and pathological features of elderly patients with hepatocellular carcinoma (HCC)and the prognosis after radical resection.Methods From January 2013 to December 2014,98 elderly patients with primary liver cancer were enrolled in this study,with 120 non-elderly patients with primary liver cancer serving as the control group.Comparison was made concerning clinical and pathological characteristics,short term postoperative outcomes and long-term prognosis between the two groups.Results The average age of patients in the elderly group(68.4±3.7)was significantly higher than in the control group(53.6 ±5.3),and the difference was statistically significant(t=23.376,P＜0.001).The positive rate of HBsAg in the elderly group was 38.8 ％,higher than in the control group (70.0 ％),and the difference was statistically significant (x2 =21.341,P＜0.001).The incidence of liver failure in the elderly group was 4.1％,higher than in the control group (0.0 ％),and the difference was statistically significant (xe =4.990,P =0.026).There was no significant difference in survival rate at 6 months,1 year and 2 years between two groups (x2 1.427,2.127,and 0.510,each P＞0.05).There was no significant difference in recurrence rate between the two groups at 1 year and 2 years(x2 =0.205 and 0.706,each P＞0.05).Conclusions Elderly patients with hepatocellular carcinoma present favorable clinical and pathological features and show similar short and long-term outcomes,compared with non-elderly patients.Radical resection is valuable in the treatment of hepatocellular carcinoma in elderly patients.

Bacterial Infections ; microbiology ; therapy ; Humans ; Integrative Medicine ; Male ; Microbial Sensitivity Tests ; Middle Aged ; beta-Lactamases ; isolation & purification

Objective:To investigate the role of SDF-1/CXCR4 in metastasis of renal cell carcinoma and to observe the intracellular location of different CXCR4 segments in renal carcinoma cells.Methods:The potential nuclear localization sequences of different CXCR4 were discovered by nuclear localization software and experiments.Full length and truncated forms of CXCR4 were fused with green fluorescent protein pEGFP-N1 and their influence on subcellular localization was examined by confocal microscopy after transfecting them into renal carcinoma cell line A498.Results:Analysis with PSORT Ⅱ Prediction revealed that the nuclear localization sequence region of CXCR4 was located between amino acids 146 and 149(RPRK).Expression products of the recombinant plasmids with SDF-1 stimulation,including EGFP-CXCR4(1-510 bp),EGFP-CXCR4(1-765 bp) and wild-type EGFP-CXCR4,were mainly located in the cell nuclei.However,expression product of EGFP-CXCR4(1-267 bp) with SDF-1 stimulation was mainly located in the renal cell cytoplasm.Expression product of wild-type EGFP-CXCR4 full length plasmid without SDF-1 stimulation was mainly located in the cell cytoplasm;these results accorded with the results of bioinformatics analysis.Conclusion:Nuclear localization sequence of CXCR4 is located in the amino acids 90 to 170,which provides a theoretical basis for further clarifying the nuclear localization sequences of CXCR4 in renal cell carcinoma cells and for finding new potential target for inhibiting the metastasis of renal cell carcinoma.

Objective To study the effects of carboxymethylated chitosan (CMCS) to nitric oxide (NO)-induced apoptosis on rat chondrocytes,and explore p38MAPK signal transduction pathway in the process and its mechanism.Methods The rat articular cartilage cells were cultured in vitro,collagen type-2 (collagen-2) immunohistochemical staining was used to identify the cartilage cells.The model of chondrocyte apoptosis was built by different concentrations of sodium nitroprusside (SNP) induction.The cells were divided into the control group,the SNP treated group SNP+CMCS treated group,and the SNP+p38 MAPK inhibitor SB203580 treated group.The apoptotic rate of chondrocytes was calculated by FCM,apoptotic nuclei was identified by Hoechst33342 stain,the mitochondrial membrane potential changes was detected by Rhodamine123 (Rho123) stain,the expression of p38 and p-p38 were detected by Western blotting analysis.Results 1-3 mmol/L SNP could induce chondrocyte apoptosis,the apoptotic rate was increased with the SNP increasing,the most obvious apoptosis was occurred in 3 mmol/L SNP treated chondrocytes,which was 69.8％ (P＜0.05).SNP could increase the nuclear fragmentation of chondrocytes,the cells with nuclear fragmentation was significantly higher than that in the control group.SNP could reduce mitochondrial membrane potential in chondrocytes,which decreased significantly compared with the control group.SNP could increase the p-p38 expression in chondrocytes,which was 4.3 times compared to the control group.CMCS of different concentrations could reduce the apoptotic rate of SNP-induced chondrocytes,which was 51.0％,29.9％ and 15.2％,which was decreased significantly (P＜0.05) when compared with 3 mmol/L SNP induced group,CMCS decreased the cells number of SNP-induced nuclear fragmentation.CMCS increased the mitochondrial membrane potential in SNP-induced chondrocytes.CMCS reduced the expression levels of p-p38 in SNP-induced chondrocytes.Conclusion CMCS has protective effect on SNP-induced apoptosis of chondrocytes.This process is completed by inhibiting the activity of p38 MAPK signal pathway.

BACKGROUND:Studies have shown that 26RFa plays an important regulatory role in bone formation, pain, endocrine, cardiovascular disease and energy metabolism. OBJECTIVE:To observe the effects of 26RFa on the proliferation and differentiation of human bone marrow mesenchymal stem cells. METHODS:In order to obtain the most efficient concentration of 26RFa, human bone marrow mesenchymal stem cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. cells were inoculated into 6-wel plates and then divided into two groups:experimental group treated with 10-11 mol/L 26RFa and control group with no 26RFa. After 8, 12 and 16 days of osteogenic induction, alkaline phosphatase activities in induced cells were detected using alkaline phosphatase kit. After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining were performed. The number of calcified nodules over each coverslip was calculated, and the expression of cbfa1 protein was detected by western blot assay. RESULTS AND CONCLUSION:After 8, 12, and 16 days of osteogenic induction, the alkaline phosphatase activities were higher in the experimental group than the control group (P<0.05, P<0.01, P<0.05). After 21 and 28 days of osteogenic induction, alizarin red staining and Von Kossa staining showed that the number of calcified nodules was higher in the experimental group than the control group, and the expression of cbfa1 protein was also higher in the experimental group (P<0.05). These findings indicate that 26RFa can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells under appropriate culture conditions.

Objective To evaluate the predictive value of plasma atrial natriuretic peptide (ANP) level on prognostic of patients with systemic inflammatory response syndrome by dynamic monitoring ANP levels.Methods Ninety-eight patients admitted to the intensive care unite were classified into survival group(n =78) and death group (n =20).The level of plasma ANP,procalcitonin,C-reactive protein and lactic acid were measured.Acute Physiology and Chronic Health Evaluation (APACHE) Ⅱ score were recorded.Results The plasma ANP level of patients in the death group was 0.40 (0.26) μg/L,significantly higher than that in the survival group(0.22(0.12) μg/L,P =0.000).Along with treatment scheme,the plasma ANP level decreased in survival group,and there were significant difference among three times (0.22 (0.12) μg/L,0.17 (0.09) μg/L,0.13 (0.11) μg/L,P =0.000).But there was no difference in ANP level of patients in death group along with the disease developing (0.38 (0.30) μg/L,0.39 (0.23) μg/L,0.39 (0.22) μg/L,P =0.99).ICU hospitalized time in survive group associated with APACHE Ⅱ score,ANP and PCT(r =0.735,0.628,0.487 respectively,P =0.000,0.001,0.021).Conclusion ANP is proved to be a good clinical index in prognostic evaluation of patients with sepsis.

Objective To evaluate the preventative effects of rehabilitation training(RT)on deep venous thrombosis(DVT)after arthroplasty.Methods Fifty-six patients with articulatio coxae or knee arthroplasty were randomly divided into a control group and an experiment group(E group).RT,including active movement of the foot and ankle,isometric contraction of the quadriceps fexoris and deep breathing training,was administered to the E group after arthroplasty.Negative cheirapsis was applied in the control group.Peak and average blood flow velocities (PABFVs)in the femoral vein,as well as DVT,were detected and measured using color ultrasound Doppler imaging before and 7 d after arthroplasty.Results PABFVs in the E group were higher than those in the control group (P

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism

OBJECTIVETo explore the expression of p-p38 MAPK protein and the number of astrocytes expressing p-p38 MAPK in CA1 hippocampus in rats during the induction of brain ischemic tolerance induced by intermittent hypobaric hypoxia (IH) preconditioning.

METHODSThirty healthy adult male Wistar rats were randomly divided into 6 groups (n = 5 in each group): sham 0 min group, IH + sham 0 min group, sham 7 d group, IH + sham 7 d group, Ischemia (Is) 7 d group, and IH + Is 7 d group. Neuropathological evaluation was performed by thionine staining in CA1 hippocampus in rats. The expression of p-p38 MAPK in CA1 hippocampus was observed by immunohistochemical staining. And the number of astrocytes expressing p-p38 MAPK was observed by immunofluorescent double labeling.

RESULTSThe results showed that IH preconditioning induced brain ischemic tolerance successfully. At the same time, IH preconditioning obviously up-regulated the expression of p-p38 MAPK protein in CA1 hippocampus, and also increased the number of astrocytes expressing p-p38 MAPK.

CONCLUSIONIt might be concluded that IH preconditioning induced brain ischemic tolerance by up-regulating the expression of p-p38 MAPK protein in pyramidal neurones and astrocytes.

Animals ; Astrocytes ; enzymology ; pathology ; Brain Ischemia ; enzymology ; pathology ; Disease Models, Animal ; Hippocampus ; enzymology ; Hypoxia ; Ischemic Preconditioning ; methods ; Male ; Phosphorylation ; Pressure ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; metabolism