Objective To explore the mechanisms of protective effect of benazepril in the treatment of experimental cyclosporin- induced chronic nephrotoxicity. Methods Rats were on low-salt diet and cyclosporine A (CsA) was administered by gastric gavage at a dose of 30?mg/kg once daily for 28 days. The expression of mRNA for intrarenal transforming growth factor-?1 (TGF-?1) and renin was detected by reverse transcription-polymerase chain reaction (RT-PCR). Intrarenal expression of TGF-?1 and Collagen Ⅳ was determined by immunohistochemistry. The effects of benazepril on these changes were also evaluated. Results Chronic cyclosporine-induced nephropathy may be related to TGF-?1 and renin mRNA up-regulation as well as matrix proteins accumulation in interstitium. Benazepril could reduce TGF-?1 mRNA up- regulation and decrease intrarenal matrix proteins accumulation. Conclusion Decreased CsA-related TGF-?1 up-regulation expression and accumulation of matrix proteins in the kidney may be related to mechanisms of protective effect of benazepril in the treatment of cyclosporin-induced chronic nephrotoxicity.

To observe the growth, expansion and differentiation of the cultured bone marrow stroma cell (BMSC) from Macaca Lrus, BMSC isolated from adult Macaca Lrus were cultured with the culture medium confected by ourselves and were induced with some cytokines such as LIF and bFGF. The results showed that the BMSC could proliferate and generate Nestin positive clones when they were cultured in vitro. After subculture, these cells could grow rapidly and differentiated into neuron like cells and astrocyte like cells further, which expressed GFAP or NSE antigen respectively. Therefore, these BMSC possess renewal and differentiation abilities. On the other hand, the culture method we used in this experiment is suitable for culture of BMSC in vitro. The BMSC might be used as the seed cells of the neural stem cells.

To develop the methodology to purify and culture expand mesenchymal stem cells (MSC) from human bone marrow and investigate the optimal system for MSC differentiation into chondrocytes mononuclear cells were harvested by gradient centrifugation on Percoll at density of 1 073g/ml and seeded in low glucose DMEM containing 10% fetal calf serum. The purity of cells was evaluated by flow cytometric technique. MSC of 2 passages thereafter were absorbed into a biomaterial of gelatin sponge and induced to differentiate in to chondrocytes for one week under the influence of TGF ? and other inductive agents. The feature of chondrocytes in engineered tissues was identified by toluidine blue staining at various time points. The results showed that human mesenchymal stem cells culture expanded were positive for CD166, CD29, and CD44, but negative for CD34, CD45, and HLA DR. Furthermore, when treated cells absorbed into the biomaterial were implanted subcutaneously into BALB c nude mice, they formed cartilage like tissues after 4 weeks.Our conclusions is that cultured marrow MSC have unique biological features and the capacity to differentiate into chondrocytes in vivo , so they are useful for cartilage engineering by serving as the seed cells.

ObjectiveTo investigate the effect of NF-?B on the differentiation and maturation of mu rine bone marrow-derived dendritic cells (DC) in vitro.MethodsThe activity of NF-?B in dendritic cells was blocked by oligodeoxyribonucleot ide Decoys (ODN Decoys) containing two special binding sites for NF-?B. Morpho logical changes of DS were observed. The expression of assistant molecules on th e cell surface was detected by using flow cytometer, and the stimulating activit y of allogeneic T lymphocyte proliferative response was determined in culture mu rine DC. The effect of NF-?B on the maturation and immunobiological activity o f murine DC was studied.ResultsNF-?B ODN Decoys were efficiently incorporated by DC, markedly suppressed the maturation of DC, the expression of assistant costimulatory molecules (CD80, CD8 6 and CD40) on the surface of DC and the secretion of IL-12, blocked the develo pment of DC and this blocked function was not reversed by lipopolysaccharide (LP S). In mixed lymphocyte reactions, DC treated with NF-?B ODN Decoys could indu ce allogeneic T cells hyphoresponsiveness, and this was associated with the inhi bition of Th1-type cytokines (IL-2 and IFN-?) production.ConclusionsNF-? B is a key gene to the development and maturation of DC. Specific interference w ith NF-?B in DC using ODN Decoys approaches could offer a novel strategy for i nducing and generating tolerogeneic immature DC and provide a promising means to induce transplant immune tolerance.

Objective To investigate the effect of murine dentritic cells vaccine modified with IFN-? inducible protein-10 gene on CTL induction. Methods DC was propagated from bone marrow (BM) of mice and pulsed with mouse prostate cancer cell line RM-1's whole lysate (Tuly-DC).IP-10 DNA fragments were inserted into pcDNA3.1(+) vector to construct recombinant plasmid IP-10/pcDNA3.1.Tuly-DC was transfected with IP-10/pcDNA3.1 by DOTAP liposome.Reversal transcript-polymerase chain reaction (RT-PCR) was used to evaluate gene transfer efficiency and chemotaxis assay was used to estimate the tansfected DC's chemotactic activity on T cells.Antitumor activity of the DC vaccine was studied in vitro by using Mixed leukocyte reaction (MLR) and cytotoxic assay (MTT assay). Results The IP-10 plasmid vector was successfully transfected into DC,which was confirmed by RT-PCR.The DC tranfected with IP-10 gene was capable of synthesizing and secreting IP-10 chemokine,which could increase the preferential chemotaxis of DC to T cells.MLR showed that the DC pulsed with whole tumor lysate and modified with IP-10 gene (IP-10/ Tuly-DC) could induce T cell proliferation significantly compared with other groups (P

Objective To study the therapeutic effects and mechanisms of Astragaloside-Ⅳ (ASG-Ⅳ) on diabetic cardiomyopathy (DCM) in the rat diabetic model. Methods Forty SD(Sprague-Dawley)healthy rats were used. The diabetic retinopathy rats model were induced by STZ, 45 mg/kg, 3d. Another 10 were injected the same amount of citrate buffer as a control group. Fasting blood glucose was measured with SureStep Plus detector 72 h later. The blood glucose of the diabetes model was ≥16.7 mmol/L. And 12 weeks later, DCM rats were divided into 4 groups randomly in the experiment, includes: DCM, ASG-Ⅳ-L (10 mg/kg), ASG-Ⅳ-M (30 mg/kg), ASG-Ⅳ-H (60 mg/kg)groups. After give dugs 4 weeks, the phosphokinase isoenzyme (CK-MB) and LDH level were tested, the cardiac hypertrophy was evaluated by HW/BW and LVW/BW. Activity of Na+-K+-ATPase and Ca2+-ATPase were determined in left ventricular tissues by ATPase ELISA Assay Kit. The content of FFA was measured and myocardial pathological examination was performed. Results Compared with the control group, blood and urine glucose were higher than experimental animal in diabetic model group, were significantly increased (P<0.05). LDH and Phosphokinase isoenzyme (CK-MB) level were significantly increased, the HWI and LVWI ratio were enhanced in DCM group (P<0.05). ASG-Ⅳ could reduce the ratio of HWI and LVWI, decrease the level of CK-MB and LDH, improve the pathomorphological changing of DCM rat model (P<0.05). Moreover, compared with DCM group, ASG-Ⅳ could restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduced the content of FFA (P<0.05). Conclusion ASG-Ⅳ plays therapeutic effect on diabetic cardiomyopathy might via restore the Na+-K+-ATPase and Ca2+-ATPase activity, reduce the content of FFA, protect the myocardial energy metabolism in DCM.

Objective To study the therapeutic window of rapamycin(RPM) concentration in primary recipients of renal transplantation. Methods An open label, multi center study was performed. One hundred primary renal allograft recipients with cadaveric donors were enrolled from 4 transplantation centers in China. The immunosuppressive regimen was triple therapy,i.e.RPM combined with CsA and steroid. A loading dose of RPM 6 mg/d was administered within 48 hours after transplantation, then a maintaining dose of 2 mg/d was administered. The whole blood concentration of RPM was measured by HPLC method. Results The whole blood concentration of RPM in this group was (6.65?2.75)ng/ml, the 10th and 90th percentile for RPM concentration was 3 2 ng/ml and 10 26 ng/ml,respectively.9 5%(8/84)patients suffered from acute rejection during the 6 month period after transplantation in this study, and the concentration of RPM in these was lower than that in non rejection patients(P=0.001). Hyperlipidemia and liver dysfunction were the most frequently adverse events, and RPM concentration was significantly associated with the concentration of triglyceride. Conclusions 4～8 ng/ml is a suitable level for RPM concentration. Regular drug monitoring and reasonable dose modulation may increase the validity and security of RPM.

To investigate the antitumor effect of herpes simplex virus thymidine kinase (HSV-TK) gene/ganciclovir (GCV) system on human bladder cancer cells (T24), a retroviral vector with the gene (pLXSN-TK) was transduced into the packaging cell line PA317. A nude mouse model with human T24 was established to examine the in vivo efficacy. The animals were randomly assigned to two treatment groups and two control groups. Treatment I and Treatment II were given in situ injection of virus suspension and PA317/TK respectively, followed by treatment with GCV for 14 days. Control I and Control II were given in situ injection of same volume of normal saline and PA317/TK respectively, followed by treatment with GCV and with normaly physiologic saline respectively for 14 days. The weight and the volume of tumor were measured. HSV-TK mRNA expression was determined by hybridization in situ. Cell apoptosis was evaluated by flow cytometry (FCM) and termininal deoxynucleofidyl transferase-mediated dUTP nick end labelling (TUNEL) technique. The results showed: (1) In vivo, the retrovirus transferred HSV-TK gene can be transduced into human bladder cancer cell T24. The tumors in T24 mice with TK gene transduced were much smaller than those in other groups. (2) After treatment with HSV-TK/GCV, the phenomenon of bladder ceancer cell apoptosis was more conspicuous as compared with that of other groups. Therefore, HSV-TK/GCV system can suppress the growth of T24 in vivo and may relate to "bystander effect". It could be a valuable therapy for human bladder cancer.
Animals ; Antiviral Agents ; therapeutic use ; Ganciclovir ; therapeutic use ; Gene Transfer Techniques ; Genetic Therapy ; Herpesvirus 1, Human ; enzymology ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Thymidine Kinase ; genetics ; Tumor Cells, Cultured ; Urinary Bladder Neoplasms ; genetics ; pathology ; therapy

Objective:To explore the precise layered and tension-reducing sutures for skin pigmented mole surgery to promote tissue healing and reduce scar hyperplasia.Methods:From January 2019 to December 2021, the First Department of Surgery of the Civil Aviation General Hospital and Tenth Department of the Plastic Surgery Hospital of the Chinese Academy of Medical Sciences treated 56 patients with skin pigmented moles aged 18-52 years, with an average age of 26 years, including 30 males and 26 females. All patients in this group underwent surgical resection of skin pigmented moles, which reached the subcutaneous fat layer. The dermis and subcutaneous tissue under the skin incision were precisely buried and guided suture by using the middle common hole equal-chord and equal-arc buried guide suture with scale marks on both ends of the needle tip.Results:The incision width of skin tissue defect in this group of patients was less than 30 mm. After the suturing was completed, the tension between the tissues on both sides of the incision and the close-fitting of each layer of tissue on both sides of the incision without dead space were realized immediately. 55 cases achieved primary incision healing. After two years of follow-up observation, there was no scar hyperplasia, and the effect was satisfactory. In only one case, local incision was red and swollen due to suture reaction, and a small amount of scar hyperplasia appeared later.Conclusions:This submerged guided suture method is an effective surgical technique for reducing skin incision scars, and it is more suitable for small incisions with a skin incision length of less than 10 mm, which is difficult to achieve layered suture of the deep tissue of the incision with ordinary suture needles.

Objective:To compare the ability of deep convolutional neural network-crop (DCNN-C) and deep convolutional neural network-whole (DCNN-W), 2 artificial intelligence systems based on different training methods to dignose early gastric cancer (EGC) diagnosis under magnifying image-enhanced endoscopy (M-IEE).Methods:The images and video clips of EGC and non-cancerous lesions under M-IEE under narrow band imaging or blue laser imaging mode were retrospectively collected in the Endoscopy Center of Renmin Hospital of Wuhan University, for the training set and test set for DCNN-C and DCNN-W. The ability of DCNN-C and DCNN-W in EGC identity in image test set were compared. The ability of DCNN-C, DCNN-W and 3 senior endoscopists (average performance) in EGC identity in video test set were also compared. Paired Chi-squared test and Chi-squared test were used for statistical analysis. Inter-observer agreement was expressed as Cohen′s Kappa statistical coefficient (Kappa value).Results:In the image test set, the accuracy, sensitivity, specificity and positive predictive value of DCNN-C in EGC diagnosis were 94.97%(1 133/1 193), 97.12% (202/208), 94.52% (931/985), and 78.91%(202/256), respectively, which were higher than those of DCNN-W(86.84%, 1 036/1 193; 92.79%, 193/208; 85.58%, 843/985 and 57.61%, 193/335), and the differences were statistically significant ( χ2=4.82, 4.63, 61.04 and 29.69, P=0.028, =0.035, <0.001 and <0.001). In the video test set, the accuracy, specificity and positive predictive value of senior endoscopists in EGC diagnosis were 67.67%, 60.42%, and 53.37%, respectively, which were lower than those of DCNN-C (93.00%, 92.19% and 87.18%), and the differences were statistically significant ( χ2=20.83, 16.41 and 11.61, P<0.001, <0.001 and =0.001). The accuracy, specificity and positive predictive value of DCNN-C in EGC diagnosis were higher than those of DCNN-W (79.00%, 70.31% and 64.15%, respectively), and the differences were statistically significant ( χ2=7.04, 8.45 and 6.18, P=0.007, 0.003 and 0.013). There were no significant differences in accuracy, specificity and positive predictive value between senior endoscopists and DCNN-W in EGC diagnosis (all P>0.05). The sensitivity of senior endoscopists, DCNN-W and DCNN-C in EGC diagnosis were 80.56%, 94.44%, and 94.44%, respectively, and the differences were not statistically significant (all P>0.05). The results of the agreement analysis showed that the agreement between senior endoscopists and the gold standard was fair to moderate (Kappa=0.259, 0.532, 0.329), the agreement between DCNN-W and the gold standard was moderate (Kappa=0.587), and the agreement between DCNN-C and the gold standard was very high (Kappa=0.851). Conclusion:When the training set is the same, the ability of DCNN-C in EGC diagnosis is better than that of DCNN-W and senior endoscopists, and the diagnostic level of DCNN-W is equivalent to that of senior endoscopists.