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

Acute Disease ; Adolescent ; Asthma ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; RNA, Viral ; genetics ; Respiratory Syncytial Viruses ; genetics ; Respiratory Tract Infections ; immunology ; virology ; Reverse Transcriptase Polymerase Chain Reaction

Objective Little research has been done on the risk factor analysis of BK virus(BKV) infection in renal transplant recipients in Chinese population.The article aimed to investigate BKV infection and analyze its risk factors in renal transplant recipients in China.Methods Renal transplant recipients who had received the detection of BKV DNA in urine and blood samples in Nanjing General Hospital from June 2015 to July 2016 were selected, while the patients with uremia hemodialysis and healthy living donors were included as control group.According to the detection results of BKV DNA in urine and blood samples, renal transplant recipients were divided into BKV DNA positive group(n=89, positive urine or blood and urine BKV DNA) and BKV DNA negative group(n=359, negative blood and urine BKV DNA).Analysis was made on BKV infection in renal transplant recipients in order to investigate the effects of factors including clinical condition, postoperative complications and immunosuppressive regimen on BKV infection.Results The positive rate of BKV DNA in urine samples of renal transplant recipients was 19.9%, which was higher than those of patients with dialysis and healthy living donors(6.3% and 4.2% respectively, P<0.001).Multivariate logistic regression analysis showed BKV infection was associated with pulmonary infection(OR[95%CI], 3.468[1.227-9.802];P=0.019) , acute rejection (OR[95%CI], 2.645[1.142-6.127];P=0.023), and FK506 (OR[95%CI], 2.408[1.104-5.254];P=0.027).Conclusion The incidence of BKV infection in renal transplant recipients increases significantly.Pulmonary infection, acute rejection and FK506-based immunosuppressive regimen are risk factors leading to BKV infection.

Background Studies showed that macophenolic acid (MPA)down-regulates and inhibits the expression and secretion of tissue growth factor and inflammatory factor,and further impacts the proliferation and inflammation process.Pterygium is an inflammatory and proliferative lesion.Whether MPA has an inhibitory effect on pterygium is unclear.Objective This study was to investigate the antifibrotic effects of macophenolic acid on pterygium fibroblasts(PFBs) in vitro and discuss its mechanism.Methods Pterygium tissue was obtained from pterygium patient during the surgery.PFBs were cultured using explants and identified with vimentin immunohistochemisty.0,0.125,0.250,0.500,1.000 μmol/L MPA were added into the culture medium,respectively,and the cells were cultured in the medium without MPA as the control group.MTT colorimetry was used to find the optimization effective concentration of MPA and evaluate their inhibitory effect on PFBs,and BrdU fluorescence staining was used to assess the growth statue of PFBs.Expressions of nuclear factor-κB(NF-κB),p65 and inhibitor of NF-κB-α(IκB-α) in the cells were detected by Western blot.Results The cells was spindle in shape 3 days after cultured and showed the vortex and radial arrangement with the positive response to vimentin.With the increase of MPA,the proliferative value of PFBs (A560)showed gradually decline,with a significant difference among the five groups (F =42.874,P＜0.01).In addition,the proliferative value of PFBs (A560) significantly lowed as the prolong of MPA active time(F=26.038,P＜0.01).BrdU fluorescence staining showed a significant decrease of DNA synthesis of PFBs with the elevation of MPA dose among the five groups(F=175.279,P＜0.05),and the A560of PFBs DNA synthesis in different concentrations of MPA groups was lower than that of the control group (all at P＜0.05).No apoptotic and necrotic cell was found after MPA action by DAPI staining.The expression level of p65 in the PFBs was 0.886±0.072 and 1.542±0.124 in the MPA group and the control group,indicating a declined value in the MPA group(P＜0.05).However,the expression value of IκB-α in the cytoplasm PFBs was significantly higher in the MPA group compared with the control group(2.141 ±0.305 vs.1.559±0.267) (P＜0.05).Conclusions MPA has an inhibitory effect on the growth of PFBs,which probably is related to the arresting of NF-κB pathway.

Objective To analysis and determine the possibility of the Citellus undulatus infected with Yersinia pestis surviving the winter in an experimental study, and to provide scientific experimental basis for the study on the mechanism of Yersinia pestis preservation. Method In 2006,09 to 2007,04 and 2007,09 to 2008,04 in Xinjiang Wusu-Gurtu natural foci of plague, under natural conditions, the over the winter process of Citellus undulatus carrying the plague bacteria was simulated, and 178 Citellus undulatus were infected with Yersinia pestis (1×107 Bacteria/mouse) using artificial injection method. One hundred seventy-eight Citellus undulatus infected with Yersinia pestis were kept into a construction of the black (1-5 ℃) basement (2 meters under the ground) in the plague focus. In doing so, these Citellus undulatuses almost simultaneously stepped into hibernation. After waking up from hibernation in following year in April, the survived mice carrying the plague bacteria were observed. Results Sixty-eight mice survived among the 178 infected with Yersinia pestis after 6 months of hibernation (through October to the following year in April), and the remaining 110 were all dead without pulling through the hibernation period. The survival rate was 38.2% (68/178). The organ culture of Yersinia pestis of the 110 dead mice(Citellus undnlatus) were tested, 67 were negative(-), 43 positive(+), with a positive rate of 39.1%(43/110). Among the rats with positive plague bacteria, the congestive pulmonary edema and the pathological changes of the hemorrhagic inflammation of the heart, liver, spleen, kidney and injection site could be seen clearly; the plague-free mice were not found to have any pathological changes. The survived 68 mice over the winter were autopsied and observed after being fed up for 20 days. No any pathological changes were found among these mice, and culturing of Yersinia pestis of the heart, liver, spleen, lungs and the tissue of injection site of these mice were all negative (-). Conclusions Citellus undulatus can carry Yersinia pestis during hibernation, but some fail to carry the bacteria through the entire process of hibernation persistently. Yersinia pestis was negative in the survived mice at the end of hibernation. The results showed that Citellus undulatus can not carry Yersinia pestis over the winter.

Objective Sirolimus ( SRL) is a potent immunosuppressive drug used to prevent acute allograft rejection after re-nal transplantation.Nevertheless, the occurrence of proteinuria has recently been recognized among patients treated with SRL-based therapy.The aim of this study was to investigate the therapeutic effect of tripterygium wilfordii hook F ( T II) on proteinuria caused by SRL in renal transplant recipients who were treated by trilogy immunosuppressive therapy of sirolimus combined with mycophenolate and hormone. Methods 52 recipients were divided into 2 groups randomly:TⅡgroup (n=27) and valsartan group (n=25).The TⅡgroup was administered 1 mg/kg/d, and the valsartan group 80-160 mg/d for consecutive 12 months.Based on primary trilogy immu-nosuppressive therapy of sirolimus combined with mycophenolate and hormone, the dosage of sirolimus was adjusted according to the target concentration 6-10 ng/ml( ELASA approach) and mycophenolate was administered 750 mg twice per day, adjusting dosage ac-cording to the mycophenolate AUC 0-12 level(35-45 mg· h/L).The evaluation of therapeutic effect includes: complete remission, proteinuria decreased by>50%; partial remission, proteinuria decreased by 20% to 50%; ineffective, proteinuria decreased by<20%. Results During the 12 month follow-up, the total effective rates in the TⅡgroup and the valsartan group were 95.2%and 86.7%respectively, in which the TⅡ group decreased more significantly (P<0.01).The total cholesterol level and triglyceride level in TⅡgroup were obviously lower than those in valsartan group(P<0.01). The total cholesterol level and triglyceride level in valsartan group increased ([6.60±0.2]mmol/L vs [7.11±1.13]mmol/L, [2.47± 1.48]mmol/L vs [2.49±0.32] mmol/L).The serum protein level in TⅡ group was obviously higher than that in valsartan group ([41.1±1.2]g/L vs [37.9±4.2]g/L, P<0.05).At 3 month, 6 month and 12 month follow-up, the average serum creatinine levels in TⅡgroup were obviously lower than those in valsartan group ([1.5±0.4]mg/dl vs [1.6±0.3]mg/dl, P<0.05), ([1.3±0.3]mg/dl vs [1.8±0.5]mg/dl, P<0.05), ([1.1±0.4]mg/dl vs [2.1±0.5]mg/dl, P<0.05).The incidence rate of adverse reaction in valsartan group was obviously greater compared with TⅡgroup( P<0.05) . Conclusion Both tripterygium wilfordii multiglycosides and valsar-tan can reduce proteinuria caused by SRL in renal transplant patients,while the application of TⅡhas more remarkable effect.

BACKGROUND: Both Allergic rhinitis and bronchial asthma are allergic diseases of respiratory tract. They frequently coexist in the same patients.OBJECTIVE: To study the incidence of allergic rhinitis in patients with bronchial asthma in Nanjing, and to analyze the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.DESIGN: The questionnaire survey.SETTING: Department of Otorhinolaryngology-Head and Neck Surgery and Department of Respiratory, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.PARTICIPANTS: A total of 134 patients with bronchial asthma including 73 males and 61 females were selected form Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from February 2001 to April 2005. Their ages ranged from 3 to 72 years. Severity of asthma of all patients was classified based on Global Initiative for Asthma (GINA). The diagnostic criteria of allergic rhinitis were established referred to the guideline of Haikou Meeting (November, 1997). All patients provided the confirmed consent.METHODS: The questionnaire survey of 134 bronchial asthma patients from the city of Nanjing was conducted. Their age, classification, severity, treatment status, and the family history of all 134 patients were investigated in details. Differences of age, age at onset, and course of disease between rhinitis and asthma of all patients were examined by two independent t tests. The correlation between clinical characteristics of allergic rhinitis and bronchial asthma was analyzed by Spearman rank correlation.MAIN OUTCOME MEASURES : Incidence of allergic rhinitis in patients with bronchial asthma and the correlation of clinical characteristic between allergic rhinitis and bronchial asthma.RESULTS：Of all 134 bronchial asthma patients，82(61．2％,82/134)were accompanied with allergic rhinitis．There were 45 males and 37 females．Of 82 patients with allergic rhinitis accompanying with bronchial asthma，severity was classified into grade Ⅰ (mild pause,n=56)，grade Ⅱ (mild persistence，n=21)，grade Ⅲ (moderate persistence，n=4)，and grade Ⅳ (severe persistence，n=1)．In addition,allergic rhinitis was classified into periodicity (n=65) and persistence(n=17)．According to the classification and seventy system recommended in ARIA,63 patients had mild symptoms and 19 had moderate and severe symptoms．Severity of allergic rhinitis was positively related to that of bronchial asthma(r=0.689,P＜0．01)．CONCLUSION: The morbidity of allergic rhinitis combining with bronchial asthma is high in Nanjing, China. While, their classification and severity are highly coherent.

BACKGROUND:Chitosan is a kind of natural biomaterial and is characterized by great biocompatibility, progressive degeneration and absorption and excellent mechanical property; however, whether it may become an ideal cytoskeleton in the engineering of cartilage tissue or not should be researched further.OBJECTIVE: To observe the hydrophilicity and adsorptivity to human nasal septum chondrocytes and the effect of its function of a novel scaffold made by [poly (dl-lactide-co-glycolide)] (PLGA)/chitosan nonwoven cloth embedded with lecithin (LEC) and poly-L-lysine (PLYS).DESIGN: Blank control study.SETTING: Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from October 2005 to June 2006. Chitosan nonwoven cloth was provided by Hainan Xinlong Company. The mainly technical parameters were detailed as the follows: degree of deacetylation ≥ 90% and relative molecular weight 2-5 × 105. PLGA/chitosan nonwoven cloth scaffold was made in Department of Otolaryngology-Hean and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA and High Polymer Institute of Sun Yat-sen University. The mainly technical parameters were detailed as the follows: mole ratio between monome lactide and glycolide 75:25, porosity 82%-86%, pore diamater 100-300 μm, shear strength 48 MPa, depth 1.5 mm and completely degenerated duration 14-18 weeks. Human nasal septum chondrocytes were the septal cartilage of nose which was derived from operated patients with deflection of nasal septum under sterile condition.METHODS: PLGA/chitosan nonwoven cloth was sheared into pieces with the size of 1.5 cm × 1.5 cm, dipped in 0.01 volume fraction of LEC anhydrous alcohol and 1 g/L PLYS for 6 hours, dealt with ultraviolet radiation after dehydration for 1hour, dipped in 0.75 volume fraction of ethanol for 24 hours, washed with Hanks solution, and incubated for 24 hours. After operations mentioned above, two kinds of novel scaffolds containing various components were obtained, and they were simple scaffold and scaffold embedded with LEC and PLYS. Cells derived from the third generation of human nasal septum chondrocytes were used to make suspension. In addition, cell suspension was grown on those two scaffolds to determine the degree of hydrophilicity through observing diffused degree of cell suspension. Whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not were observed under phase contrast microscope so as to determine adsorptivity between cells and scaffolds; meanwhile, growth of cells and production of matrix were also observed.MAIN OUTCOME MEASURES: ① Hydrophilicity of those two kinds of scaffolds (diffused degree of cell suspension of human nasal septum chondrocytes on scaffold) and adsorptivity to cells (whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not); ② growth of cells and production of matrix.RESULTS: ① When simple scaffold was put in cell suspension of human nasal septum chondrocytes, cell suspension showing like balls attached to the surface of scaffold, and then, scattered into space of scaffold gradually. During the period of culture, phase contrast microscope indicated that masses of cells attached to the surface of fiber of scaffold.When the petri dish was shaken, cell groups drifted irregularly. The adherent rate was (21±3.7)%. With the cultured time passing by, matrix was not produced. ② When scaffold embedded with LEC and PLYS was put in cell suspension of human nasal septum chondrocytes, cell suspension scattered into space of scaffold rapidly. The adherent rate was (89±5.6)%, which was higher than that of single scaffold group. This suggested that scaffold showed a strong hydrophilicity.Phase contrast microscope indicated that chondrocytes as the form of monome or community were distributed between scaffold and fiber averagely and attached to the surface of fiber of scaffold. When the petri dish was shaken, cell groups did not drift irregularly, and only a few of cells were scarred at the bottom of petri dish. This suggested that scaffold had a strong adsorptivity to cells. One week after culture, matrix showing like cobweb was produced among fibers of scaffold.With the cultured time passing by, matrix was produced abundantly.CONCLUSION: The novel scaffold of PLGA/chitosan nonwoven cloth embedded with LEC and PLYS is characterized by an excellent hydrophilicity and adsorptivity to human nasal septum chondrocytes; meanwhile, it also can secrete matrix.

AIM To develop an HPLC method for the simultaneous content determination of five constituents in Chairen Capsules (Bupleuri Radix,Ziziphi spinosae Semen,Chuanxiong Rhizoma,etc.).METHODS The analysis of 80％ methanol extract of this drug was performed on a 30 ℃ thermostatic Merck Purospher STAR C1s column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 237,290 and 335 nm.RESULTS Spinosin,ferulic acid,cinnamaldehyde,liquiritin and ammonium glycyrrhizate showed good linear relationships within the ranges of 0.009 26-0.092 6 μg (R2 =0.999 6),0.056 2-0.562 μg (R2 =0.999 8),0.130-1.30 μg (R2 =0.999 8),0.275-2.75 μg (R2 =0.999 5) and 0.568-5.68 μg (R2 =0.999 2),whose average recoveries were 100.73％,100.62％,100.02％,100.41％ and 99.96％ ％ with the RSDs of 2.20％,1.37％,1.52％,1.30％ and 0.80％,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Chairen Capsules.

Objective BK virus-associated nephropathy ( BKVAN) after kidney transplantation is a key factor that influence the prognosis of transplant kidney .To our knowledge , it is believed to be associated with immune suppression .We observed the cura-tive effect and influencing factorsof anti-rejection scheme that Leflunomide was administered instead of Mycophenolate Mofetil ( MMF) on transplant kidney BKVAN .. Methods This study included 15 kidney transplant recipients with pathologically confirmed BKVAN in Nanjing General Hospital of Nanjing Military Region form March 2007 to March 2013 .Leflunomide was administered instead of Myco-phenolate Mofetil ( MMF) .Serum creatinine level , renal allograft loss rate and side effects of leflunomide were monitored after medica-tion switch.The patients were divided into two groups , which were renal allograft loss group and renal allograft survival group , for fur-ther analyses . The differences between each groups in clinical characteristics as well as histochemical features of the transplanted kidneys were analyzed to determine the cause of renal allograft loss in patients with BKVAN . Results Six patients experienced renal al-lograft loss after switching to leflunomide and needed hemodialysis , and 9 patients had stable renal allograft function , renal allograft loss rate was 40.0%.Hyperuricemia occurred in 8 patients in the period before the medication switch and in 5 patients after the switch;a decrease in blood white cell orplateletcount was found in 2 patients during both periods;an increase in Alanine aminotransferase ( ALT) level occurred in one patient after the medication switch .There were no statistically significant differences in any of the above parame-ters before and after the medication switch.Compared to allograft survival group, serum creatinine level[(1.80 ±0.53)mg/dL vs (2.74 ±0.58)mg/dL, P=0.007], the number of B lymphocytes [(206.44 ±144.96) vs (439.67 ±267.77), P=0.047] and CD68[(588.44 ±271.80) vs (944.67 ±259.32), P=0.025] in renal allograft tissue were significantly higherin the allograft loss group. ConclusionLeflunomide is a safe and effective medication for BKVAN .Patients with significantly increased serum creatinine level might have a poorer prognosis .Significantly increased B lymphocytes and CD 68 cells in renal allograft tissue might indicate a poor prognosis.