OBJECTIVETo investigate the expressions of sphingosine-1-phosphate receptors 1-3 (S1P1- 3) in the corpus cavernosum of castrated male rats and its relationship with the NOS/NO/cGMP and RhoA/Rho kinase signaling pathways.

METHODSWe equally randomized 18 eight-week-old healthy male SD rats into a sham-operation control, a castration, and a testosterone replacement (TR) group and harvested the bilateral testes and epididymides from the rats in the latter two groups, followed by 4 weeks of subcutaneous injection of testosterone propionate at 3 mg per kilogram of the body weight per day for those in the TR group and that of plant oil for those in the control and castration groups. At the age of 12 weeks, we measured the serum testosterone (T) level and maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP) of the animals and determined the expressions of SlP1-3, eNOS, P-eNOS, ROCK1, and ROCK2 in the corpus cavernosum by Western blot and immunohistochemistry.

RESULTSThe serum T level was significantly decreased in the rats of the castration group as compared with those of the control and TR groups ([0.41 ± 0.04] vs [16.01 ± 1.02] and [15.84 ± 1.32] nmol/L, P < 0.01), with no statistically significant difference between the latter two groups. The ICPmax/MAP at 0 V, 3 V, and 5 V electric stimulation was remarkably lower in the rats of the castration group (0.088 ± 0.014, 0.323 ± 0.014, and 0.432 ± 0.012) than in those of the control group (0.155 ± 0.011, 0.711 ± 0. 010, and 0.819 ± 0.024) and TR group (0.153 ± 0.012, 0.696 ± 0.017, and 0.763 ± 0.027) (P < 0.01), with no significant difference between the latter two groups. With GAPDH as internal control, the animals of the castration group showed markedly reduced expressions of S1P1 ([49.99 ± 3.39]%), eNOS ([46.82 ± 3.81]%) , and P-eNOS ([45.42 ± 4.35]%) in comparison with those in the control group ([72.57 ± 3.06], [89.76 ± 3.98], and [82.53 ± 8.92] and TR group ([71.77 ± 4.43], [87.19 ± 4.23], and [79.82 ± 7.38]%) (P < 0.01) , while the expressions of S1P2, S1P3, ROCK1, and ROCK2 were significantly upregulated in the castration group ([82.35 ± 4.13], [61.03 ± 5.14], [74.50 ± 4.02], and [69.83 ± 5.75]%) as compared with those in the control group ([41.67 ± 1.68], [31.66 ± 2.67], [35.69 ± 5.56], and [39.85 ± 7.17]%) and TR group ([42.80 ± 3.87], [32.25 ± 4.22], 38.06 ± 5.21], and [42.36 ± 4.44]%) (P < 0.01).

CONCLUSIONAndrogen deficiency induces significant reduction of ICPmax/ MAP in male rats, which is possibly associated with the decline of S1P1 in the corpus cavernosum, inhibition of the eNOS/NO/cGMP signaling pathway, increased expressions of S1P2 and S1P3, and activation of the RhoA/Rho kinase signaling pathway.

Animals ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Orchiectomy ; Penis ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Lysosphingolipid ; metabolism ; Testosterone ; blood ; pharmacology ; rho-Associated Kinases ; metabolism

Objective To investigate the diagnostic value of laboratory evaluation of renal injury in early diagnosis index. Methods Eighty-six patients in in-patient and out-patient of the department of urology were tested in serum C (Cys C), serum creatinine (Cr) and serum UREA nitrogen (UREA). According to the diagnostics (sixth edition), we evaluated their values for diagnosis of renal injury by analyzing the characteristics of the work curve (ROC). Results According to the renal function indexes, 3 groups were divided such as group A (normal renal function control Ccr 80 mL/min) or the Cys C, Cr, and UREA in (0.47 ± 0.24) mg/L, (85 ± 14) μmol/L, and (4.55 ± 1.33) mmol/L, group B (a decrease in renal function reserves, and renal insufficiency 20 mL/min or less Ccr < 80 mL/min) of Cys C, Cr, and UREA in (1.59 ± 1.29) mg/L, (214 ± 173) μmol/L, and (11.04 ± 8.24) mmol/L and group C (renal failure and uremia Ccr < 20 mL/min) of Cys C, Cr, and UREA in (3.47 ± 0.75) mg/L, (669 ± 466) μmol/L, and (21.22 ± 13.10) mmol/L respectively. As reducing in Ccr, Cys C, Scr and UREA concentration increased and correlated with Ccr negatively. According to the mazimum Youden index, the critical values of Cys C, Cr, and UREA were 0.90 (mg/L), 115 (μmol/L) and 7.33 (mmol/L) separately, the sensitivity (%), specificity, positive predictive value (%), negative predictive value (%), diagnostic accuracy (%) and the area of the ROC curve (AUC) were Cys C (77.3%, 97.6%, 97.0%, 81.1%, 77.3%, 0.908), Cr (65.9%, 97.6%, 96.5%, 74.1%, 65.9%, 0.817), and UREA (63.6%, 97.6%, 96.4%, 72.8%, 63.6%, 0.785) respectively. The diagnostic accuracy showed that the Cys C > Cr > UREA. Their correspounding ROC curve (AUC) were in the area of 0.908, and 0.785 and 0817. Cys C had the highest AUC (0.908), and achieved a good diagnosis effect. Conclusion Renal damage laboratory indexes of in the early diagnosis, Cys C has a higher sensitivity and specificity to be worth of clinical promotion.

Objective To explore the epidemiology,etiology and prevention strategy of early respiratory infections (≤1 month) in lung transplantation recipients with donation after cardiac death donors.Method The clinical data of donors and recipients,particularly on early respiratory infections,were retrospectively analyzed in 17 lung transplantations.Result From Jan.2015 to Apr.2015,12 episodes of early respiratory infections (≤ 1 month) in 17 lung transplantation recipients occurred (12/17,70.6％).The organisms most frequently involved were bacteria:Pseudomonas aeruginosa (4/26,15.4％),Klebsiella pneumoniae (4/26,15.4％),Staphylococcus aureus (3/26,11.5％),and Acinetobacter baumannii (3/26,11.5％).Of 26 bacterial strains,3 were Methecillin resistant Staphlococcus aureus,3 were carbapenem resistant Acinetobacter baumanni,2 were carbapenem resistant Pseudomonas cepacia,2 were extended spectrum b-lactamase-producing Klebsiella pneumoniae,and one was carbapenem and quinolone resistant Pseudomonas aeruginosa.Conclusion The morbidity of early infections is high in lung transplantation recipients.In our experience,bacterial respiratory infections are most common in the early post-transplant period (≤ 1 month).Incidence of Aspergillus spp.and Cytomegalovirus pneumonia is lower than before lung transplantation,probably due to the spread of universal prophylaxis.

Objective To explore the value of combined use of laboratory indicators for diagnosis of early renal functional damage. Methods Eighty-six patients with various kidney diseases were enrolled in the Second Affiliated Hospital of Guangzhou Medical University. On admission,the serum Cystatin C(Cys C),creatinine(Cr), Urea,etc were determined. The value of using combined laboratory indicators in the diagnosis of renal functional damage was obtained through the analysis of the receiver operating characteristic curve(ROC curve);multiple variable indicators were grouped to establish multiple logistic regression models to be compared and evaluated. Results In the early and late renal injury groups(group B of 32 cases and group C of 12 cases),the serum levels of Cys C,Cr, and Urea were significantly higher than those in the normal renal function control group(group A of 42 cases),the elevation in level in group C being the most significant〔Cys C(mg/L):3.47±0.75 vs. 1.59±1.29,Cr(μmol/L):669±466 vs. 214±173,Urea(mmol/L):21.22±13.10 vs. 11.04±8.24,P<0.05 or P<0.01〕. The areas under the ROC curve(AUC)made by combinations of Cys C,Cr and Urea for the diagnosis of renal damage were 0.908,0.817 and 0.785. In the four different kinds of combination of the indicators,the AUC were sequentially arranged from large to small area as follows:Cys C+Cr+Urea=Cys C+Cr>Cys C+Urea>Cys C>Cr+Urea(0.920=0.920>0.911>0.908>0.809). In this sequence,the AUC made by Cys C+Cr+Urea and Cys C+Cr were equal,both 0.920, whose sensitivity was 75.0%,specificity 100.0%,positive predictive value 100.0%,negative predictive value 80.0%and diagnostic accuracy rate 87.5%. So,Cys C + Cr combination could be used to substitute Cys C + Cr + Urea, and the former clinical diagnostic effect was the best,much higher than that by using AUC whose curve was made by Cys C alone. Conclusion The value of using only one laboratory indicator for diagnosis of patients with early renal functional damage is not high,while applying Cys C+Cr combination can improve the diagnostic effect greatly,and its sensitivity and specificity are higher.

BACKGROUND: During xenogenic liver transplantation, major histocompatibility antigen can induce immunological rejection, and immunosuppressant can cause adverse effect on organism. Recently, treatment prior to transplantation induces immune tolerance, which is perspective for organ transplantation.OBJECTIVE: To investigate the correlation between thymus induction and immunological rejection during liver transplantation. METHODS: A total of 40 male SD rats of clean grade were selected as donors. Moreover, 30 male Wistar rats of clean grade and 10 male SD rats of clean grade were selected as recipients. The donor rats were divided into allogeneic gene transplantation, allotransplantation, cyclosporine, and thymus induction groups, with 10 rats in each group. The modified Kamada and improved two-cuff technique was used to establish a stable rat orthotopic liver transplantation model. The cyclosporine group was given cyclosporine (50 mg/kg) for 5 successive days. Thymus induction group was injected with major histocompatibility antigens (50 pL) for 5 successive days. Other groups were not given any interventions. Survival time of rats was recorded in each group. Pathological observation and mixed lymphocyte cultured were performed at days 3, 7, 14, 21, and 28 after transplantation. RESULTS AND CONCLUSION: Survival time was longer in the thymus induced group compared with other groups (> 60 days), damaged level was mild, local immunological rejection was reduced, and lymphocytes were decreased. The effect after liver transplantation was similar to allogeneic gene transplantation but superior to cyclosporine intervention (P < 0.05). This suggested that thymus induction relieved immunological rejection following liver transplantation.

BACKGROUND: Most patients who underwent liver transplantation would suffer acute rejection or transplanted liver failure resulted by chronic rejection, therefore, inducing specific immune tolerance via varied pathways is the ideal method to solve this problem. OBJECTIVE: To treat rat transplanted liver by injecting transforming growth factor β1 (TGF-β_1) plasmid, and to analyze the relationship between TGF-β1 and allograft rejection from gene level. METHODS: A total of 30 male, Wistar rats were served as allogenic liver donors, and 10 male, SD rats served as syngeneic donors Totally 40 male SD rats were served as liver recipients, and divided into 4 groups by order number table: ailogenic transplantation, syngeneic transplantation, ciclosporin, and ciclosporin plus TGF--β_1 groups. In each group, rat orthotopic liver transplantation model was established by modified Kamada and improved two-cuff technique. After modeling, rats were received cyclosporine 1-5 days in the cyclosporine group, or intraperitoneal injected ciclosporin for 1-5 days, combined with TGF-β_1 plasmid 0-2 days in the cyclosporine plus TGF-β_1 group. No intervention was performed in the other groups. The survival time of rats were recorded, and the pathological changes was detected at days 3, 7, 14, 21, and 28 after transplantation, then the mixed lymphocyte culture was performed. RESULTS AND CONCLUSION: The survival time of rats in syngeneic transplantation group and cyclosporine plus TGF-1,β_1 group was more than 60 days, which was obviously greater than that of allogenic transplantation and cyclosporine groups (P< 0.05). The histopathologic slide showed that there was moderate and severe acute rejection, with evident intrahepatic inflammatory cell infiltration in the allogenic transplantation and cyclosporine groups. Few rejections were observed in the syngeneic transplantatior group, which was close to the normal lever tissues. Mixed lymphocyte culture of the cyclosporine plus TGF-β_1 group was superior to the syngeneic transplantation group or cyclosporine group (P < 0.05). The results demonstrated that cyclosporine combined with local injection of TGF-β_1 plasmid can relieve post-transplant immune rejection.

BACKGROUND: A small number of mesenchymal stem cells (MSCs) present in bone marrow, which would gradually drop with age. OBJECTIVE: To verify the effect of adherent method on culture of bone marrow-derived MSCs. METHODS: Under anaesthesia, bone marrow cells were obtained from femur and tibia of rats, cultured by DMEM containing calf serum, placed in an incubator containing 5% CO_2 at 37 ℃. The culture medium was renewed after 24 hours, and remained periodical medium change with once per week. The weakly adherent cells were passaged. The cell morphology, growth curve, and the expression of cell-surface markers were identified by flow cytometry and immunocytochemical staining. RESULTS AND CONCLUSION: After 24 hours of culture, the cells could adhere to the walls with fusiform or triangle shapes, proliferated faster after 2-3 days, and presented whirlpool-like or clustering. The cells reached a logarithmic growth phase after 2 days, and into the late stationary phase after 12 days, which covered the bottle after 15 days. The cultured cells were positive to CD90 and CD54. The results verified that bone marrow-derived MSCs can be isolated by adherent method. This method is easy operation, and can maintain cell activity preferably.

Objective: To optimize the extraction technology of Panax notoginseng and Scrophulariae radix from Rupixiao granule.Methods: With the dry extract rate and transfer rates of ginsenoside Rg1, ginsenoside Rb1 and harpagoside as the comprehensive index, the orthogonal design was adopted to investigate the effects of the amount and concentration of ethanol, extracting duration and times on the extraction technology.The contents of ginsenoside Rg1, ginsenoside Rb1 and harpagoside were determined by HPLC.Results: The optimal extraction technology was extracted twice with 8-fold amount of 60% ethanol with 2 h per time.The transfer rate of ginsenoside Rg1, ginsenoside Rb1 and harpagoside was (79.4%±1.56%), (42.62%±0.68%) and (44.89%±0.58%)(n=3), respectively.The dry extract rate was (20.99%±0.411%).Conclusion: The optimized extraction technology is stable and feasible, which can be used for extracting Panax notoginseng and Scrophulariae radix from Rupixiao granule.

Objective To investigate the diagnostic value of VCA-IgA and multilayer enhanced spiral CT(MESCT) in the diagnosis of nasopharyngeal carcinoma.Methods Retrospective analysed the data on serum VCA-IgA test and MESCT of 385 patients with suspected nasopharyngeal disease,and the results were confirmed by surgery and pathology.Comparatively analysed the two methods in differential diagnosis of nasopharyngeal carcinoma and benign diseases.Finally,calculated the sensivity,speficity,positive predictive value,negative predictive value and accuracy in diagnosis of nasopharyngeal carcinoma.Results The positive rate of serum VCA-IgA,MESCT and combination of those two methods in naospharyngeal carcinoma were significantly higher than those in nasopharyngeal benign disease(P<0.05);combination of the methods significantly improve the sensitivity and accuracy in diagnosis of nasopharyngeal carcinoma,the specificity was lower than VCA-IgA,higher than MESCT alone.Conclusion Combination of serum VCA-IgA and MESCT were excellent strategy for diagnosis of nasopharyngeal carcinoma.

Objective To explore the correlation between resistins rs2161490and rs1423096 genotype with type 2 diabetes mellitus (T2DM)in Guangdong.Methods Collected 178 blood of newly diagnosed T2DM in the Second Affiliated Hospital of Guangzhou Medical University from January 2015 to November 2015 as the patient group and 192 blood of healthy physi-cal examination as the control group.Analysis of the two groups of gene distribution frequency was to reach the genetic equi-librium,comparative two gene loci frequencies of resistin rs2161490 and rs1423096 in case group and control group was sta-tistically significant,and compared the distribution frequency of rs2161490 locus T→C and rs1423096 locus A→G between the patient group and the control group.Then made a logistic regression analysis:analysing the risk two loci each genotype of resistin rs2161490 and rs1423096 to T2DM,adjust of the gender and age,and the changes of the risk of the two variables. Comparative blood lipids biochemical indexes between case group and the control group,mode the correlation analysis be-tween TG,CHOL,HDL-C and LDL-C levels of serum lipids in patients with rs2161490 and rs1423096 each genotypewere performed.Results The sample was consistent with Weinberg Hardy’s law of inheritance,which was representative of the population,comparing two gene loci frequency of resistin rs2161490 and rs1423096 of case group and control group:com-parinng CT,TT,CC of rs2161490 genotype,there was no statistically significant difference (P=0.834,>0.05),and com-parinng AA,AG,GG of rs2161490 genotype,there was no statistically significant difference (P=0.960,>0.05).Each gen-otypes with T2DM risk analysis,there was no statistically significant difference(P>0.05).Adjusting the risk change after the two variables,gender and age,there was no statistically significant difference (P>0.05);TG,CHOL,HDL-C and LDL-C in each of the genes expression levels correlation analysis,there was no statistically significant difference (P>0.05).Con-clusion Analysis results showed that the frequency of two loci all genotypes in the case group and control group were no statistical significance (P>0.05).The risk of two loci gene type of rs2161490 with rs1423096 and type 2 diabetes were be-fore and after the covariate adjustment had no statistical significance (P>0.05 ).Each genotype of rs2161490 with rs1423096 and lipid levels had no statistical significance (P>0.05).Thus infer that two genotypes is not risk for type 2 dia-betes genes in guangdong area.