Objective To investigate the effects of DNMT1 on neuropathic pain behavior and neuropathic pain modulation.Methods Thirty-two male SD rats, weighing 200-250 g, were randomly assigned into 4 groups (n =8 each):sham operation group (group S),chronic constrictive injury group (group CCI),CCI+ DNMT1-siRNA group (group CDS),CCI+ control-siRNA group (group CCS).Group CDS were intrathcally injected of DNMT1-siRNA (2 μg/10 μl),and group CCS were intrathcally injected of control-siRNA 7,8,9 days after operation.Mechanical withdrawal threshold (MWT)and thermal withdrawal latency (TWL)were measured before operation and on day 3,5,7,9,12,14 after operation.The rats were then sacrificed and L4-L6 segments of the spinal cord were removed for determination of SOCS1,p-ERK,p-CREB expression using Western blot on day 14.Results Compared with group S,MWT and TWL in group CCI and CCS were significantly decreased on day 3,5,7,9,12,14 after operation (P <0.05).Compared with group CCS,MWT and TWL in group CDS were significantly increased on day 9,12,14 after operation (P < 0.05 ). Compared with group S and CDS,SOCS1 was significantly downregulated,p-ERK and p-CREB were significantly upregulated in group CCI and CCS (P <0.05 ).Conclusion Intrathcal injection of DN-MT1-siRNA significantly relieves neuropathic pain by upregulating SOCS1,downregulating p-ERK and p-CREB in rats spinal cords.

Objective: To explore the effect of different processing methods on the content changes of ferulic acid and total flavonoids in wheat bran and its processed products by HPLC and UV spectrophotometry.Methods: Ferulic acid was determined by HPLC with SHIMADZU C18 (250×4.6 mm,5 μm) as the analytical column, the mobile phase was composed of acetonitrile-0.1% phosphoric acid solution (16∶84), the flow rate was 1.0 ml·min-1, the detection wavelength was 320 nm and the column temperature was 35℃.Total flavonoids was determined by UV spectrophotometry at the wavelength of 258 nm(±2 nm).Results: The content of ferulic acid in the crude bran, the toasted to brown bran, the toasted to dark brown bran, the over toasted bran and the honey toasted bran was 2.07, 2.33, 1.68, 1.11 and 0.83 μg·g-1, and total flavonoids was 5.96, 7.15, 9.73, 13.61 and 9.42 mg·g-1, respectively.Conclusion: The higher the toasted processing degree is, the lower content of ferulic acid and the higher content of total flavonoids are in wheat bran.

Objective To explore the anti-tumor effects of Egr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy in mice bearing H22 hepatocarcinoma and its mechanism. Methods The recombinant plasmid pcDNAEgr-IFNγ mixed with liposome was injected into tumor. 48 h later, 370 kBq 125I-UdR was injected into tumor. The tumor growth rates at different times were observed. After 3 d gene-radionuclide therapy, the concentration of IFNγ in cytoplasm of H22 cells and cytotoxic activities of splenic CTL of the mice in different groups were examined. Results The tumor growth rates of pcDNAEgr-IFNγ +125 I-UdR group were obviously lower than those of control group, 125I-UdR group and pcDNAEgr-1 +125I-UdR group 6-15 d after gene-radionuclide therapy. IFNγ protein was found in cytoplasm of H22 cells in PcDNAEgr-1FNγ+125I-UdR group after 3 d gene-radionuclide therapy. Cytotoxic activity of splenic CTL in pcDNAEgr-IFN7 + 125I-UdR group was significantly higher than that in the other groups (P<0.01). Conclusions The anti-tumor effects in vivo of pcDNAEgr-IFNγ gene therapy combined with 125I-UdR radionuclide therapy are better than those of 125I-UdR therapy.

OBJECTIVETo perform a meta-analysis on clinical outcomes of minimally invasive percutaneous plate osteosynthesis (MIPPO) or open reduction and internal fixation (ORIF) for distal tibial fractures in adults.

METHODSPubmed database (from 1968 to March 2014), Cochrane library and CNKI database (from 1998 to March 2014) were searched. Case-control study on minimally invasive percutaneous plate osteosynthesis (MIPPO) or open reduction and internal fixation (ORIF) for distal tibial fractures in adults were chosen,and postoperative infection, operative time, blood loss, fracture nonunion rate, delayed union,fracture malunion rate were seen as evaluation index for meta analysis. The system review was performed using the method recommended by the Cochrane Collaboration.

RESULTSTotally 5 studies (366 patients) were enrolled. Meta-analysis showed that there were significant meaning in postoperative infection between MIPPO and ORIF [OR = 0.23,95% CI (0.06,0.92), P = 0.04]; fracture nonunion rate in MIPPO was lower than in ORIF group [OR = 0.16, 95% CI (0.03,0.76), P = 0.02]; operative time in MIPPO was shorter than in ORIF group, and had significant difference [MD = -14.42, 95% CI (-27.79, -1.05), P < 0.05]; blood loss in MIPPO was less than in ORIF group [MD= -87.17,95%CI (-99.20, -75.15), P < 0.05]; there was no obviously meaning in delayed union between two groups.

CONCLUSIONFor distal tibial fractures in adults, MIPPO has, advantages of short operative time, less blood loss, lower incidence of infection and fracture non-uniom, but with high fracture malunion rate. MIPPO for distal tibial fractures in adults is better than ORIF, and the best treatment should choose according to patient's condition.

Bone Plates ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Minimally Invasive Surgical Procedures ; methods ; Operative Time ; Tibial Fractures ; surgery

Objective To observe the expression level of macrophage stimulating protein (MSP) in acute on-chronic liver.failure (ACLF) patients,and to explore the clinical significance and correlation with different immune regulatory factors.Methods The double antigen sandwich enzyme-linked immunosorbent assay method was used to detect MSP in the peripheral blood of 45 patients who were diagnosed with ACLF and 32 cases of chronic hepatitis B (CHB).The MSP levels were compared among ACLF patients with different outcomes,and the MSP level of healthy person was used as control.Meanwhile,liver function and hepatitis B virus (HBV) load were detected,and the expressions of peripheral blood CD4+ interferon (IFN)γ+ (helper T cell 1 Th1),CD4+ interleukin (IL)-4+ (helper T cell 2,Th2),CD4+IL-17+ (helper T cell 17,Th17) and CD4+ CD25+ Foxp3+ (regular T cell,Treg) were measured by flow cytometry.The comparison of means between two samples was done by t test,and oneway ANOVA and linear correlation analysis were also used.Results The serum MSP levels in ACLF patients,CHB group and healthy control were (1.65±0.46) ng/mL,(1.43±0.32) ng/mL and (1.23±0.21) ng/mL,respectively.The serum MSP level in ACLF patients was significantly higher than both CHB patients (t=2.163,P=0.035) and healthy control (t=4.032,P=0.01).The MSP level in ACLF survival group was statistically higher compared with ACLF death group ([2.29 ± 0.42] ng/mL vs [1.42±0.17] ng/mL,t=1.973,P=0.042).Th2,Th17 cells in ACLF group,CHB group and healthy control group were (1.51±0.27) ％ and (1.94±1.02)％,(0.42±0.08)％ and (0.55±0.36)％,(0.23±0.19) ％ and (0.26±0.19) ％,respectively,which were all significantly different (F=7.759 and 37.229,respectively;both P＜0.01).The MSP level was positively associated with the number of Th2 (r=0.386,P=0.032) and Th17 (r=0.644,P=0.000),and the ratio of Th17/Treg (r =0.605,P=0.000);while it was negatively associated with the number of Th1 (r=-0.212),Treg (r=-0.262) and the ratio of Th1/Th2 (r=-0.394) (all P＞0.05).Conclusion MSP is involved in the progress of ACLF,and it may be associated with clinical outcomes and cellular immune imbalance of ACLF patients.

Objective:To establish the quality standard for Huayu pills. Methods:TLC was adopted to identify salvia miltiorrhi-za, rhubarb charcoal and panax notoginseng; the contents of tanshinoneⅡA , chrysophanol and physcion in Huayu pills were deter-mined by RP-HPLC with Wondasil C18 (250 mm × 4. 6 mm,5μm) as the analytical column, the mobile phase was composed of metha-nol-0. 1% phosphoric acid solution (85 ∶15), the flow rate was 1. 0 ml·min-1,the detection wavelength was 254 nm, the column temperature was 35℃, and injection volume was 10 μl. Results:The TLC spots were clear and well-separated without any negative in-terference; the calibration curve of tanshinoneⅡA , chrysophanol and physcion was linear over the range of 0. 212-2. 12 μg ( r =0. 999 9),0. 159-1. 59 μg(r=0. 999 9) and 0. 072 6-0. 726 μg(r=0. 999 9), the average recovery was 99. 55 %, 99. 56% and 100. 60%, and RSD was 1. 57%,2. 26% and 2. 28%(n=6), respectively. Conclusion: The method is fast and accurate with good specificity, and can be used for the quality control of Huayu pills.

Objective:To establish the quality standard for Qingchang mixture.Methods:TLC was adopted to identify angelica sinensis,paeoniae radix and phellodendri cortex.HPLC was used to determine the content of ferulic acid on a Wondasil C18 (250 mm×4.6 mm,5 μm) analytical column,the mobile phase was composed of acetonitrile-0.1% phosphoric acid solution(17∶83),the flow rate was 1.0 ml·min-1, and the detection wavelength was 316 nm and the column temperature was 30℃.Results:The TLC spots were clear and well-separated without any negative interference.The calibration curve of ferulic acid was in good linearity over the range of 39.6-316.8 μg·ml-1(r=0.999 9),the average recovery was 98.75% and RSD was 1.21%(n=9).Conclusion:The method is fast and accurate with good specificity,which can be used for the quality control of Qingchang mixture.

Objective To Investigate the influence of high-intensity pulsed electromagnetic field (HIPEMF) on neural differentiation of neonatal rats neural stem cells in vitro.Methods Neural stem cells (NSCs) were isolated from the subventricular zone of 3-day-old neonatal rats and cultured with serum-free condition medium for 14 days.All the NSCs were then randomly classified into an experimental group which received the stimulation of HIPEMF (0.1 Hz,4 T,8 pulses) and a control group which received no special intervention.Differentiation of the culture was induced by addition of 10％ fetal bovine serum on the first day after intervention,the morphological changes of cells were observed under the microscope at different time points.The NSCs adhered to the wall and differentiated for seven days,the immunofluorescence was employed to observed and calculate the ratio of differentiated cells with astrocyte marker GFAP or neuronal markers TUJ1.RT-PCR and western blotting were used to measure the expression levels of the differentiated cells based on gene and protein levels,respectively.Results Immunofluorescence staining showed the number of TUJI positive cells in the experimental group(33.4％ ± 5.1％)was significantly more than the control group (26.5％ ± 7.0％),while the number of GFAP positive cells was decreased(23.9％ ± 5.0％) as compared with the control group(36.2％ ± 2.2％).RT-PCR showed that the TUJ1 mRNA expression levels in the experimental group was (1.682 ± 0.086) times of the control group.Western blot showed that the expression of TUJ1 (0.729 ±0.061) in the experimental group was higher than in the control group (0.590 ± 0.157),while the expression of GFAP in the experimental group (0.566 ± 0.056) was less than in the control group(1.034 ± 0.051).Conclusions HIPEMF facilitates differentiation of neural stem cells to neurons,at the cost of reducing astrocytic differentiation.

The present study was aimed at the comparison of the pharmacokinetics of pure chlorogenic acid and extract of Solanum lyratum Thunb. The animals were allocated to two groups, and were administered chlorogenic acid or extract of S. lyratum Thunb. at a dose of 50.0 mg/kg orally. Blood samples were collected up to 8 h post-dosing. Plasma chlorogenic acid analyses were performed using an HPLC method with UV detector. The pharmacokinetic parameters were evaluated using non-compartmental assessment. Significant differences existed in the two groups for AUC0-t, AUC0-∞ and CLz/F. The reliable HPLC method was successfully applied to the determination of chlorogenic acid in rat plasma at dosage of 50.0 mg/kg.

Objective To investigate the differential diagnostic value of computed tomography (CT) combined with serum tumor markers in borderline ovarian tumors (BOT) and benign epithelial ovarian tumors (BET).Methods The CT data in 28 patients with BOT and 41 patients with BET,both confirmed by surgery and pathological,were analyzed retrospectively.Their preoperative serum carbohydrate antigen 125 (CA125),human epididymis secretory protein 4 (HE4) and carcinoembryonic antigen (CEA) detection results were collected.The CT images features and serum tumor markers levels were compared between the two groups.Results The difference in the appearance rate of tumor solid composition,thick septum and wall nodule between the two groups had statistical significance (x2 =25.135,5.240,5.066,P＜0.05).The serum CA125 level had statistical difference between the two groups (Z=3.202,P＜0.05),while serum HE4 and CEA levels had no statistically significant difference between the two groups(Z=0.330,1.122,P＞0.05).The optimal critical value,sensitivity and specificity of serum CA125 level in differential diagnosis of two kinds of tumor was 42.45 U/mL,53.6％ and 85.4％.The overall diagnostic rate of solid composition and thick septum for diagnosing the two kinds of tumor was 78.5 ％.The overall diagnostic rate of solid composition,thick septum and CA125 level for diagnosing the two kinds of tumor was 81.2％.Conclusion The appearance of solid composition,thick septum and serum CA125 level increase in epithelial ovarian tumor may help to identify BOT and BET.