[Objective]To investigate the best internal fixation methods of upper segment complicated femoral fractures(USCFF).[Method]Fifty-two cases(54 limbs) with USCFF were treated.In all patients open fracture reduction with use of internal fixation were done.Intramedullary interlocking nails(IIN) were used to treat 32 cases(34 limbs) of adult's USCFF and 130? plate were used to treat 20 cases of children's USCFF.Thirty-six limbs of them were closed fractures,and 18 limbs were opene.Measures of auto-ilium transplant(5 limbs),homolegous allograft bone transplant(10 limbs)were also taken.[Result]Fifty-two patients were followed-up from 9 to 40 months with an average of 16 months.Infection,fracture nonunions,malunion and femoral head necrosis complications were not found.The average period of union of adult and children fractures was respectively 6.8 months and 6.5 months.The long term effect was evaluated according to Ma Yuanzhang's evaluation standard,94.4 percent showed excellent function of joints and limbs.[Conclusion]Appropriate selection of internal fixation according to the age of patients and satisfactory fracture reduction are key points to improve outcome of USCFF.IIN in treating fiacture of adult and 130? plate in treating fracture of children are more ideal selections.

Objective To investigate the rate of vaginal colonization of Candida species in patients with diabetes,and to discuss the relationship between prevalence of vulvovaginal candidiasis(VVC) and diabetes mellitus(DM).Methods Genital tract examination and fungal cultures of discharge taken among 144 patients with DM(DM group) and 150 healthy subjects(control group) were performed.Chrom Agar Candida mediums were used to study the isolates of 64 strains cultured in 144 participants.Results The isolated rates of Candida species were 44.4%(64/144) in subjects with DM and 18%(27/150)in healthy subjects,the isolated rate of Candida species in DM group was prominently higher than that in control group(P

Objective To explore the effects of different doses of contrast agent (CA) on types of time-signal intensity curves (TICs) and semi-quantitative parameters after dynamic contrast-enhanced MRI (DCE-MRI) on Walker 256 murine breast tumor model.Methods A total of 12 rats burdened Walker256 breast cancer models were established and divided into 3 groups randomly, 4 in each group.Routine MR and DCE-MRI scans of the rats using Bruker Pharmascan 7T MR scanner were performed.Doses for the 3 groups were 0.2, 0.3, and 0.5 mmol/kg, respectively.MR data, TICs types and semi-quantitative parameters from each different dose group were statistically studied and compared to observe the differences.Results Tumors were enhanced significantly after injection.The types of TICs in all tumors were the Ⅲ pattern which was not influenced by CA doses.Semi-quantitative parameters of first enhancement (Efirst), maximal enhancement (Emax), washout enhancement (Ewash), and signal enhancement ratio (SER) showed statistical differences among the three dose groups (P＜0.05).Semi-quantitative parameters of time to peak (Tpeak) and washout velocity (Vwash) showed no statistical differences among the three dose groups (P＞0.05).Mean signal intensity of each group was highly negatively linear correlated with scan times after the peak (r=-0.972, P=0.000;r=-0.971, P=0.000;r=-0.989, P=0.000).The washout slope (slopewash) showed no statistical differences among the three groups (P＞0.05).Conclusions Injection doses of CA didn't change the TIC type, Tpeak, Vwash, and Slopewash.These parameters are comparable among different medical centers and can be considered as prior parameters to monitor the efficacy of neoadjuvant chemotherapy.

Aim To develop a sensitive, specific and accurate method for the pharmacokinetic study of QO-58 ( a novel M channel opener ) in rats after intragas-tric ( ig) and intravenous ( iv) administration. Meth-ods QO-58 was administered at the doses of 25,50, 100 mg · kg-1 ( ig ) and at single dose of 100 mg · kg-1(iv), respectively. Blood samples were obtained at intervals after each administration. Plasma samples were deproteinized with acetonitrile after addition of in-ternal standard, and detected by RP-HPLC. The main parameters of pharmacokinetics were calculated by DAS2. 1. 1 software. Results The calibration curve in plasma was linear over the range of 0. 1 ~160 mg · L-1 in rat plasma, and the limit of detection ( LOD) was 0. 1 mg · L-1 . The intra-day and inter-day RSD was less than 20%. The recovery of QO-58 in rat plas-ma was 89. 56% ~101. 38%. The concentration-time curves of QO-58 in rat palsma were consistent with the two-compartment model after both oral and intravenous administration. The main pharmacokinetic parameters for QO-58 following oral administration with three doses (25, 50, 100 mg· kg-1 ) in rat were as follows:Cmax (mg·L-1):8.25,16.29,18.27;T12β(h): 8.24, 5. 01, 5. 92; AUC0-∞ ( g · min · L-1 ):261. 94, 189. 57,90. 65. Conclusion The developed method is simple and specific, and is suitable for preclinical pharmacokinetic studies of QO-58 .

Objective To elucidate the effects of eicosapentaenoic acid(EPA)on the metabolic abnormalities and renal pathologic changes during the early stage of diabetic nephropathy in KKAy/Ta mice.Methods Sixteen KKAy/Ta mice(7 weeks of age)were randomly divided into two groups(n=8).Since 12 weeks of age,EPA group were injected with EPA at 1 g?kg-1?d-1 intraperitoneally for 8 weeks,and the control group were injected with 0.9% saline.The levels of serum fatty acids were detected at 20 weeks of age by gas chromatography.The phenotypic characterizations were measured at 12,16 and 20 weeks of age.Renal morphological examinations were performed after 8 weeks of treatment.Results Serum EPA levels in KKAy/Ta mice treated with EPA [(125.8?15.5)?g/mL] were significantly higher than those in control group [(69.2?7.8)?g/mL] at 20 weeks of age(P

ced glutathione in MHD patients appears to be associated with an improvement of oxidative stress.

Objective To investigate the effects of parathyroid hormone(PTH)on microinflammatory and nutritional status in maintenance hemodialysis(MHD)patients.Methods Ninety-eight MHD patients were selected,who hod undergone hemodialysis for at least three months before the study and were in a stable clinical status without signs of infection or disease activity.The serum level of intact PTH was measured by electrochemiluminescence immunoassay(ECLIA),while the serum levels of interleukin(IL)-1β,IL-6,IL-8 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).The levels of C-reactive protein(CRP),albumin(Alb),pre-albumin(PA),hemoglobin(Hb)and lipids were measured.Body measurement and modified quantitative subjective global assessment(MQSGA)was done simultaneously.Correlation analysis between serum PTH level and the parameters for inflammation and nutrition Was performed.Results The serum levels of intact PTH in MHD patients[(353.46±102.41)ng/L]were significantly higher than those in the control people[(57.45±5.76)ng/L,P<0.01],and the serum levels of IL-1β,IL-6,IL-8,TNF-α and CRP were significantly higher in MHD patients than those in the control people(P<0.01 or <0.05).Relative body weight(RBW),triceps skin fold thickness(TSF),mid-arm circumference(MAC)and mid-arm muscle circumference(MAMC)in MHD patients decreased significantly(P<0.05 or <0.01),while the score of MQSGA increased markedly(P<0.01).The levels of intact PTH showed significantly positive correlations with the levels of CRP,IL-1β, IL-6,TNF-α, lipoprotein(a) [Lp(a)] , serum phosphorus and ages of MHD(P<0.05 or <0.01 ).The levels of intact PTH showed significantly negative correlations with RBW, MAC, MAMC, Alb, Hb and total cholesterol(TC) in MHD patients (P<0.01 or <0.05) . And there was also significantly positive correlation between PTH and MQSGA in MHD patients (P<0.05). Conclusion PTH is probably involved in the presence and the progression of malnutrition-inflammation-atherosclerosis syndrome in MHD patients.

Objective To elucidate whether down-regulation of homo sapiens eukaryotic translation elongation factor 1 alpha 2 (EEF1A2) expression induces apoptosis in pancreatic cancer cells and its possible mechanisms. Methods Two siRNAs targeting human EEF1A2 were synthesized and the siRNA/liposome complexes were transfected into the pancreatic cancer cell line BxPC-3. RTPCR and Western blot were used to analyze the change of EEF1A2 expression and the apoptosis rate of BxPC-3 cells was studied using Annexin-V/PI assay. To identify the mechanisms involved, the apoptosis associated proteins such as caspase-3, caspase-8, caspase-9, PARP, cytochrome C and Bid were detected by Western blotting. Results Both EEF1A2-targeting siRNAs reduced the EEF1A2expression, and the No. 2 siRNA inhibited EEF1A2 expression to less than 25 % in mRNA and protein levels. Down-regulation of EEF1A2 expression in BxPC-3 cells enhanced cell apoptosis (15.28% ±3.65%) at a greater level than negative siRNA-expressing cells (10. 11% ± 3. 05%) or mock cells (9.41 % ±4.14 %). Furthermore, reduction of EEF1A2 activated the pro-caspase-8, pro-caspase-3,pro-caspase-9,PARP and Bid to their active forms, and increased the expression of cytochrome C.Conclusions These data suggest that EEF1A2 down-regulation could significantly induce apoptosis of pancreatic cancer cell line BxPC-3, which is likely mediated by the death receptor and mitochondrial apoptotic pathways.

Objective To explore the effect of down-regulated methionine synthase reductase (MTRR) gene on the apoptosis and autophagy pathway, and offer a possible approach for the MTRR to reverse the multi-resistant ovarian cancer. Methods (1) The experiment was divided into 3 groups, SKOV3/DDP-MTRRi (down-regulated MTRR group), SKOV3/DDP-NC (negative control group), and SKOV3/DDP (blank control group). Different concentration of cisplatin (0, 1, 2, and 4 μg/ml) treated on 3 groups cells. The apoptosis rate was measured by flow cytometry (FCM). Autophagy was detected by immunofluorescence. Autophagy microtubule associated protein light chain 3β(LC3B) and p62 were detected by western blot. The formation of autophagosome of cells was observed by transmission electron microscope. (2) Detection of autophagy and apoptosis of SKOV3/DDP-MTRRi induced by rapamycin. The experiment was divided into 4 groups included rapamycin group (5 nmol/L rapamycin), rapamycin+cisplatin group (5 nmol/L rapamycin+4μg/ml cisplatin), cisplatin group (4μg/ml cisplatin) and blank control group. LC3B and p62 protein were detected by western blot. The survival rate cells were detected by methyl thiazolyl tetrazolium (MTT) method. The apoptosis rate was measured by FCM. (3) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4 μg/ml) after 48 hours, then detecting the protein expression of caspase, Bcl-2 family in apoptosis pathway and the key proteins in phosphatidylinositol-3 kinase (PI3K)/protein kinase B (Akt) autophagy pathways by western blot, getting the time when the proteins′expression changed. Results (1) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC, and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4 μg/ml) after 48 hours, apoptosis and autophagy of 3 groups of cells were gradually increased with the increased concentration of cisplatin. The apoptosis rate of SKOV3/DDP-MTRRi cells [(26.2 ± 1.4)%] were significantly increased compared with the SKOV3/DDP-NC cells or SKOV3/DDP cells [(14.8 ± 2.4)%, (14.2 ± 2.4)%;all P<0.05] at 2μg/ml cisplatin. Immunofluorescence tests revealed that the aggregates of LC3B in SKOV3/DDP-MTRRi cells were more than that of SKOV3/DDP-NC cells and SKOV3/DDP cells. The expression of LC3B of SKOV3/DDP-MTRRi cells was lower than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The expression of p62 of SKOV3/DDP-MTRRi cells was higher than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The structure of chloroplast was integrity and autophagosome was dispersing in plastids of SKOV3/DDP-NC cells and SKOV3/DDP cells. Organelles disappear and vacuoles increased obviously in SKOV3/DDP-MTRRi cells, no autophagosome was observed. (2) The expression of LC3B of rapamycin+cisplatin group was higher than those of other 3 group cells (1.72±0.08,1.43±0.04, 1.37±0.11, and 1.11 ± 0.09;P<0.05). The expression of p62 of rapamycin + cisplatin group was significant decreased (0.58 ± 0.10,0.94 ± 0.12, 1.21 ± 0.11, and 1.57 ± 0.10; P<0.05). The survival rate of rapamycin + cisplatin group was higher than that of cisplatin group [(0.78±0.03)%vs (0.62±0.03)%;P=0.018], the apoptosis rate was significant decreased in rapamycin+cisplatin group [(59.0 ± 3.9)% vs (40.4 ± 3.0)%, P=0.019]. (3) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC, and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4μg/ml) after 48 hours, the expression of Bax in 3 groups cell were not evidently changed (P=0.661). The expression of Bcl-2 was significantly decreased in SKOV3/DDP-MTRRi cells (P=0.030). The expression of caspase-3, caspase-7, caspase-9, and poly (ADP-ribose) polymerase (PARP) were not evidently changed (P>0.05), but cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, and cleaved PARP were significantly increased in SKOV3/DDP-MTRRi cells (P<0.05). For the autophagy pathway, the expression of phosphorylated Akt (p-Akt) and phosphorylated mammalian target of rapamycin (p-mTOR) were significantly increased (P<0.05), but Akt and mTOR had no significant variation. The expression of phosphatase and tensin homologue deleted on chromosome ten (PTEN) was significantly decreased (P<0.05). Conclusions MTRR silencing significantly increase cisplatin-induced apoptosis and reduce the autophagy induced by cisplatin in SKOV3/DDP cells. Down-regulation of MTRR enhanced the chemosensitivity of cisplatin-resistant ovarian cancer cells may be by activating caspase and Bcl-2 apoptosis family and inhibiting the PI3K/Akt autophagy pathway.

Objective To study the biological effects of down-regulatingof methionine synthase reductase (MTRR) gene on cisplatin resistant ovarian cancer SKOV3/DDP cell in vitro and in vivo. Methods (1) Establishing the cell line of MTRR down-regulated. Four short hairpin RNA (shRNA) for MTRR gene (U6-GFP-Neo-homo-1106, U6-GFP-Neo-homo-1931, U6-GFP-Neo-homo-419, U6-GFP-Neo-homo-1460) were designed respectively. Western blot was used to detect the interference efficiency and selected the most efficient shRNA. The MTRR 1106 was selected as the best silencing effect of interference fragment and then therecombinant plasmid vector pSicoR-1106 was constructed and transfected into SKOV3/DDP cells.The stably transfected cells was obtained by screening of flow cytometry(FCM).Fluorescence quantitative reverse transcription (RT)-PCR and western blot were used to detect the expression of MTRR mRNA and protein. (2) Study in vitro: recombinant plasmid expression vector pSicoR-1106, pSicoR-NC and packaging plasmid were respectively transfected into 293T cell. SKOV3/DDP cells were transfected by viral supernatant. The experiment was divided into three groups, namely SKOV3/DDP-MTRRi (down-regulated MTRR group), SKOV3/DDP-NC (negative control group), and the SKOV3/DDP (blank control group). The cell growth curves and half maximal inhibitory concentration (IC50) of cisplatin were made by methyl thiazolyl tetrazolium (MTT) method. Three groups cells were treated with different concentration of cisplatin (0, 1, 2 and 4 μg/ml). The clonogenicity efficiency was observed by clony formation test. The cell cycles were measured by FCM . (3) Study in vivo: three groups cells were subcutaneously inoculated into the nude mice to develop a tumor model. Mice were injected intraperitoneally with cisplatin at 2.5 mg/kg (once every 2 days, in 21 rounds), then the tumor growth was observed. The expression of MTRR and proliferation-related Ki-67 antigen by immunohistochemistry in xenograft tumors were measured. Results (1) Results showed that U6-GFP-Neo-homo-1106 was the best shRNA with interference effect to MTRR. The recombinant plasmid pSicoR-1106 was constructed and transfected into SKOV3/DDP. The MTRR mRNA and protein were down-regulated after transfected. This result showed that MTRR down-regulated SKOV3/DDP cell line was constructed successfully. (2) The cell growth curves showed that the growth of SKOV3/DDP-MTRRi cells were significantly decreased compared with that in the SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The IC50 of SKOV3/DDP-MTRRi, SKOV3/DDP-NC and SKOV3/DDP were 4.01, 7.90, and 8.91 μg/ml, respectively. The IC50 of SKOV3/DDP-MTRRi was significantly lower than that in control cell groups (P<0.05).Clony formation tests showed that the clony numbers of varied concentration of cisplatin of SKOV3/DDP-MTRRi were significantly less than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). FCM showed that when the cisplatin concentration rose to 4 μg/ml, the G0/G1 phase cell ratio in SKOV3/DDP-MTRRi cells group was (72.8±5.0)%, which was significantly higher than those in the SKOV3/DDP-NC cells group and SKOV3/DDP cells group [(64.4±2.5)%and (64.3±3.0)%], respectively (all P<0.05).(3) Six weeks after nude mice intraperitoneal injection with cisplatin, the tumor volume of SKOV3/DDP-MTRRi, SKOV3/DDP-NC and SKOV3/DDP were respectively (97 ± 32), (168 ± 45), and (173 ± 32) mm3, the tumor weight were (0.36±0.17), (1.08±0.17), and (1.11±0.20) g, in which tumor volume and weight of SKOV3/DDP-MTRRi were significantly less than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (all P<0.05). In three groups tumor tissue, positive rates of MTRR were respectively 2/8, 5/8, and 7/8, the positive rates of Ki-67 were respectively1/8, 6/8, and 7/8, in which SKOV3/DDP-MTRRi was significantly lower those SKOV3/DDP-NC cells and SKOV3/DDP cells (all P<0.05). Conclusion The growth and cisplatin resistance of ovarian cancer cells could be decreased by down-expressing of MTRR gene in vitro and in vivo.