Objective To search for the methods preventing nephrotoxic injury of amikacin,Methods Case-control research was used in this study. There were 50 normal children in control group The urine routine, the ?2-microglobulin (?2 -M), mosmol and THP in urine and blood, the AIb, rGT and NAG in urine, the renal function and serum concentration of amikacin were determined respectively.The 43 patients with serious illness childten in study group, were divided into 2 groups (Group 1 and group 2 ). Group 1 (23 cases) was treated only with amikacin for 7 days, and group 2 (20 cases) was treated with vitaminC, vitamin E and amikacin for 7 days. Before treatment, the 3rd and 7th day during the treatment, all the items mentioncd above were examined in gtoup 1 and 2.Results The incidences of nephrotoxic injury of amikacin are 87 per cent (20/23)and 55 per cent (11/20) respectively in group 1 and 2. There is significant difference (P

Objective To investigate the effects of sirolimus and 3-methyladenine (3-MA) on the autophagy in,as well as matrix matalloproteinase (MMP)-1 and MMP-3 secretion by,human skin fibroblasts (HSFs).Methods HSFs were isolated from the circumcised foreskin of a healthy male,and subjected to primary culture.After 3 to 10 passages,HSFs were incubated with sirolimus of 20,50,100,250 nmol/L and 3-MA of 0.5,2.0,5.0,10.0 mmol/L respectively for 4 hours followed by the observation of autophagy and detection of MMP-1 and MMP-3 levels in the supernatant by enzyme linked immunosorbent assay (ELISA).Those HSFs remaining untreated or treated with dimethyl sulfoxide served as the control.Results The percentage of autophagy-positive cells was 59.075％ ± 6.884％,76.350％ ± 5.226％,85.063％ ± 6.002％,86.288％ ± 5.558％ and 96.825％ ± 1.500％ respectively in HSFs treated with sirolimus of 0,20,50,100 and 250 nmol/L; significant differences were observed between the 5 groups (P ＜ 0.01 ) but not between the cells treated with sirolimus of 50 and 100 nmol/L (P ＞ 0.05).After being treated with 3-MA of 0,0.5,2.0,5.0 and 10.0 mmol/L,the percentage of autophagy-positive cells in HSFs was 63.037％ ± 5.876％,34.425％ ± 5.183％,19.700％ ± 3.028％,12.900％ ± 3.334％ and 7.775％ ± 2.293％ respectively with a significant difference between these groups (all P ＜ 0.01 ).Elevated levels of MMP-1 and MMP-3 were observed in the supernatant of HSFs treated with sirolimus of 250 nmol/L and 3-MA of 10.0 mmol/L (all P ＜ 0.05).Conclusions The autophagy in HSFs can be upregulated by sirolimus,but downregulated by 3-MA.For the secretion of MMPs by HSFs,3-MA and high concentrations of sirolimus exert a promotive effect,and the effect of 3-MA is in a concentration-related manner,but the influences of sirolimus at lower concentrations remain unclear.

Objective To study the X-ray signs of forearm and leg in skeletal fluorosis and its diagnostic value,aim at finding the easy examination parts.Methods One thousand four hundred and forty subjects were examined using developed shield,darkroom and other portable dedicated device combined with a small X-ray machine.A total of 384 cases were diagnosed skeletal fluorosis.All patients were divided into different groups and the time,degree and range of X-ray to the forearm and calf elbow,knee,and long bone were compared.Results The X-ray change in the forearm elbow was earlier than that of the leg knee,and trabecular bone change was the earliest indicator,197 cases and 157 cases,respectively,and the difference was statistically significant (x2 =28.006,P ＜ 0.01).Membrane ossification of forearm backbone was earlier than that of the leg,and most of them were degree Ⅰ photos,213 cases and 126 cases respectively.The difference was statistically significant (x2 =17.626,P ＜ 0.01).The direction of the interosseous membrane ossification was from the forearm radius to the ulna,then to the fibula and tibia,and was accompanied by changes in the aggravation of forearm.A variety of indicators were observed,especially the membrane ossification in bone and joint trabecular bone and the long bone was the most active,and the forearm was more sensitive,obviously than that of the calf.Conclusion In the X-ray screening or detection of endemic fluorosis,the forearm radiography is a simple,economical,and effective diagnostic method.

Objective To observe the changes of autophagy in human skin fibroblast (HSF) model for photoaging. Methods HSF model for photoaging was established through repeated exposure to ultraviolet B (UVB). Those HSFs receiving no irradiation served as the control. The degree of aging was evaluated by p-galactosidase assay, and autophagy level was detected. Results After repeated exposure to UVB, most pho-toaged HSFs were deformed and distorted, and some of them even died. The percentage of P-galactosidase-positive cells was 50.60% ± 5.04% and 14.58% ± 2.69%, respectively in photoaged and control HSFs (P< 0.01). Significant difference was also observed in the proportion of cytophagosome-positive cells between photoaged and control HSFs (14.91% ± 4.59% vs 68.45% ± 8.25%, P < 0.01). Conclusion The HSF model for photoaging shows obviously abnormal appearance and stagnant growth with increased degree of senescence and decreased autophagy compared with normal control HSFs.

Objective To evaluate the safety and feasibility and clinical effectiveness of living relative donor kidney transplantation(LDKT)and summarize its clinical experience. Methods The clinical data of 30 cases of LDKT were retrospectively analyzed.Except for 2 cases being donated by spouse,the others were donated by blood relative donors.6 cases shared two haplotypes,and 22 cases shared one haplotype,and one case 4 mismatched,and 1 fully mismatched.All donors underwent open nephroectomy,in which 7 cases donated right kidneys and 23 donated left kidneys.In 30 cases of recipients,1 case received cadaver donor kidney transplantation and lost her allograft because of superacute rejection.Triple-combined immunosuppressive protocols consisted of calcineurin inhibitor (CNI),mycophenolate mofetil(MMF)or azathioprine(AZa)and steroid. Results All donors'hospital stay was 7 to 10 days postoperatively without any surgical complications. All donors kept their normal kidney function within 3 to 6 months'follow-up.Except for 1 case of death because of lung in fection,29 cases of recipients survived,in which 28 cases kept their normal function kidney within 1 to 4 years of follow-up and 1 case occurred chronic allograft nephropathy after one year.Except one case of DGF,29 cases of recipients retained their normal kidney function in 3 to 5 days postoperatively.Rejection episodes occurred in 4 cases,of which 3 cases were reversed by methylprednisone and 1 case by antithymocyte globulin(ATG)and Tacrolimus.Pneumonia developed in 3 cases,of which 2 cases were cured and 1 case failed.Hematoma was found around allograft in 1 case and wag surgi cally removed.Urinary leakage was happened in 2 cases of recepients and were cured by conservative treatment. Conclusions LDKT is safe and feasible with good long-term results and more advantages such as optimal HLA matches and less ischemia time and lower acute rejection,low-dose immunosup pressants.

AIM: To study the effects of 1.5 MAC halothane and sevoflurane on ischemic myocardium. METHODS: The isolated rat heart were perfused with halothane and sevoflurane and HR, LVEDP, LVDP, +d p /d t , -d p /d t , coronary flow (CF), the myocardial ATP content and Ca 2+ -ATPase activity were determined before and 10 min and 25 min after ischemia. In the meantime, LVP was recorded during 25 min ischemia. RESULTS: 1.5MAC sevoflurane significantly increased CF in normal isolated rat hearts. Both halothane and sevoflurane depressed myocardial contractile function, increased normal myocardial energy storage. After 10 min ischemia, the decrease of myocardial ATP content were slowed down by halothane and sevoflurane, especially halothane. During 25 min of ischemia, the onset time of contracture was significantly delayed, and the contracture intensity was alleviated by halothane, but not sevoflurane. CONCLUSION: Halothane has better protective effect on ischemic myocardium than sevoflurane through preventing the decrease of myocardial ATP content and Ca 2+ -ATPase activity during ischemia.

[Objective] To observe the effects of autophagy induced by different doses of ultraviolet B (UVB) irradiation on the apeptosis in human skin fibroblasts.[Methods] Skin fibroblasts were isolated from the circumcision specimen of a 23-year-old healthy male,and subjected to a primary culture.After 3 to 10 passages,the cells were collected and applied in the following experiment.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation of some fibroblasts treated with different concentrations (0,0.5,2.0,5.0and 10.0 mmol/L) of 3-methyladenine (3-MA).To qualitatively and quantitatively detect the autophagy in fibroblasts treated with different concentrations of 3-MA and in fibroblasts treated with 3-MA of 0.5 mmol/Lfollowing UVB irradiation,monodansylcadaverine (MDC) staining was carried out,and immunofluorescence was used to detect the expression of microtubule-associated protein 1 light chain (LC3).Some fibroblasts were classified into 8 groups to remain untreated,be irradiated with UVB of 30,50 and 100 mJ/cm2 alone,treated with 3-MA of 0.5 mmol/L alone,or treated with 0.5 mmol/L 3-MA following irradiation with UVB of 30,50 and 100 mJ/cm2,respectively,then,cell apoptosis was qualitatively detected by Hoechst and propidium iodide (PI)staining,and quantitatively detected by flow cytometry with annexin V and PI.[Results] The percentage of autophagic cells was (63.037 ± 5.876) ％ in fibroblasts treated with starvation condition,significantly decreased to (34.425 ± 5.183) ％ in fibroblasts treated with 3-MA of 0.5 mmol/L.The expression of LC3 showed a gradually increasing trend from untreated fibroblasts,to fibroblasts irradiated with UVB of 30,50 and 100 mJ/cm2,while the increase was attenuated by the 4-hour treatment with 3-MA immediately after the irradiation.Compared with the other concentrations,the 3-MA of 0.5 mmol/L showed the least influence on the viability of fibroblasts.The addition of 3-MA of 0.5 mmol/L increased the percentage of cells both positive for Hoechst and PI staining in fibroblasts irradiated with UVB of 50 mJ/cm2,but decreased that in fibroblasts irradiated with UVB of 100 mJ/cm2.Similarly,the percentage of middle and late apoptotic cells was significantly higher in fibroblasts irradiated with UVB of 50 mJ/cm2 followed by treatment with 3-MA of 0.5 mmol/L than in those irradiated with UVB of 50 mJ/cm2alone ((10.933 ± 0.839) ％ vs.(7.267 ± 0.473) ％,t =5.20,P＜ 0.05),but lower in fibroblasts irradiated with UVB of 100 mJ/cm2 followed by treatment with 3-MA of 0.5 mmol/L than in those irradiated with UVB of 100 mJ/cm2alone ( (7.100 ± 0.781 ) ％ vs.( 1 0.133 ± 0.681 ) ％,t =6.29,P ＜ 0.05 ).[Conclusion]s The irradiation with UVB of 50 mJ/cm2 may protect fibroblasts by inducing autophagy and suppressing apoptosis,while the high level of autophagy induced by UVB of 100 mJ/cm2 may lead to autophagic cell death in fibroblasts.

Objective To introduce a new technique for urethral coverage in Snodgrass hypospadias repair,and to evaluate its effectiveness and complications.Methods From April 2003 to February 2006, this new procedure was performed in 289 children with hypospadias aged 3 months to 12 years (mean age,2. 4 years).The native meatus of urethra was identified subcoronal in 78 cases,penile/shaft in 136,penoscrotal in 36 and scrotal in 16;and 23 cases had undergoneⅡstage operation and re-operation.The overlapping coverage with bilateral shaft based vascularized dartos pedicle was done in the new urethra by Snodgrass hy- pospadias repair in these children.Results All the cases were followed for 3 months to 2 years.Postoper- atively,urinary fistulas developed in 32 cases (11%).Of them,11 were cured spontaneously within 4 weeks. The incidence of actual urinary fistula was 7% (21/289).Of the 21 fistulas which were not cured,11 (5%) occurred in 214 cases of distal hypospadias;and 10 (13%) in 75 cases of proximal hypospadias,Ⅱstage and re-operation.No dehiscence and diverticulum was found.Combined with mucosal collar technique,the ventral skin of the penis was sewn on the midline.During the follow-up,excellent cosmetic results with normal-ap- pearing circumcised penis were achieved in most patients.Conclusions Bilateral shaft based vascularized dartos pedicle urethral coverage procedure is a reliable and effective method for preventing urethral cutaneous fistulas and dehiscence.This method can reconstruct a satisfactory cosmetic appearance of the penis.

Objective To evaluate effects of sirolimus(a classic autophagy inducer)and starved culture on autophagy of a human cutaneous squamous cell carcinoma cell line A431. Methods Cultured A431 cells and HeLa (a human cervical carcinoma cell line)cells were both classified into 5 groups to be treated with DMEM alone(control group), 0.1%dimethyl sulfoxide alone(DMSO group), 20 nmol/L sirolimus(20?nmol/L sirolimus group), 80 nmol/L sirolimus(80?nmol/L sirolimus group), and Earle′s balanced salt solution(EBSS group)respectively. After 4?hour treatment, Western blot analysis was performed to measure the expressions of autophagy?related markers microtubule?associated protein 1 light chain 3A/3B (LC3A/B) and recombinant gamma?aminobutyric acid receptor associated protein(GABARAP), and acridine orange staining to determine autophagy levels in these cells. Results As Western blot analysis showed, the ratio of LC3A/B?Ⅱto LC3A/B?Ⅰin A431 cells was similar between the control group and DMSO group(P > 0.05), but significantly higher in the 20?nmol/L sirolimus group, 80?nmol/L sirolimus group and EBSS group than in the control group(all P < 0.05). Western blot results from HeLa cells were similar to those from A431 cells. Bivariate correlation analysis revealed that the protein expression of GABARAP was positively correlated with that of LC3A/B ?Ⅰ in both HeLa cells(r = 0.869, 95% CI: 0.807 - 0.999, P = 0.051)and A431 cells(r = 0.837, 95% CI: -0.173 - 0.989, P = 0.037), but negatively correlated with that of LC3A/B?Ⅱ in both HeLa cells(r = -0.742, 95% CI: -0.982 - 0.406, P = 0.042)and A431 cells(r = - 0.684, 95% CI: -0.977 - 0.500, P = 0.047). Acridine orange staining showed that the percentages of autophagosome?positive A431 cells and HeLa cells were significantly increased in both the 80?nmol/L sirolimus group(23.750% ± 0.260% and 33.307% ± 0.715% respectively)and EBSS group(32.450% ± 0.488% and 66.097% ± 1.141% respectively) compared with the control group(15.987% ± 0.242% and 14.117% ± 0.295%, respectively, all P < 0.05). Conclusion The classic autophagy inducer sirolimus and starved culture can upregulate the autophagy level of A431 cells, and GABARAP may be highly correlated with LC3A/B.

Objective To study the proliferation of CRTH2 (CD4+ CD294+ Th2)cells promoted by pneumococcal vaccine through antigen presentation of dendritic cells (DCs),so as to provide a new approach for amplification and sorting of Th2 cells.Methods CDs induced from peripheral blood mononuclear cells were cocultured with T lym-phocytes after loading pneumococcal vaccine antigen,mixed lymphocyte reaction (MLR)was detected by cell count-ing kit-8(CCK8),DCs and CRTH2 cells were analyzed by flow cytometry.Results Pneumococcal vaccine could promote the maturation of DCs,together with TNF-a,it was adjuvant for maturation of DCs.Pneumococcal vaccine antigen-loaded DCs could increase the rate of subsets of CRTH2 cells on day 5([0.93±0.10]%)compared with day 1([0.70±0.02]%),and absolute number also increased (both P <0.05).Conclusion Amplification of CRTH2 cells can be greatly promoted by pneumococcal vaccine antigen-loaded DCs,which might be one of the effective way to induce amplification of Th2 cells.