Objective To investigate the mechanism of pulmonary fibrosis induced by paraquat (PQ),and the effect of Xuebijing injection in treatment of PQ poisoning.Methods Seventy-two male Wistar rats were randomly divided into control group,PQ poisoning group,and Xuebijing intervention group,with 24 rats in each group.Pulmonary fibrosis was induced by single garage at the dosage of 50 mg/kg of PQ,while 1 mL of distilled water was given by gavage in control group.Xuebijing injection at the dosage of 4 mL/kg were given intraperitoneally at 30 minutes after exposure to PQ in Xuebijing group,and it was repeated every 12 hours; same amount of physiological saline was given intraperitoneally in PQ group and control group.The experiment lasted for 14 days.Six rats in each group were sacrificed on 1,3,7,14 days,respectively,after insult,and 30 minutes after the last intervention.The lung tissues were harvested,the changes in pathology in lung tissue and the degree of pulmonary fibrosis were observed with optical microscope with hematoxylin-eosin (HE) staining and Masson stain.The ultrastructure changes in lung tissues were observed with transmission electron microscopic,and the content of hydroxyproline (HYP) in the lung tissue was determined by alkaline hydrolysis.The expression of mitochondrial fusion protein 2 (Mfn2) was determined by Western Blot.Results ① HE staining:in PQ group,inflammation was most marked on the 3rd day.On the 7th day,exudates in the alveoli started to be organized,and hypertrophic fibroblasts were seen to secrete slim collagen fibers,and fibrosis could be seen in alveoli.On the 14th day,intensive hyperplasia of fibroblasts could be observed,and the alveolar structure was destroyed and collapsed,with deposition of collagen deposited with formation of pulmonary fibrosis.At the same time,pathologic changes were milder in Xuebijing group than those in PQ group.② Masson staining:the degree of inflammation in alveoli and pulmonary fibrosis were less marked in Xuebijing group than those of PQ group on the 14th day.③ Under the transmission electron microscopy,it was found that the mitochondria of lung tissue cells was relatively less in number on the 14th day in PQ group,and the majority of them underwent degeneration,swelling and damage.Basement membrane became folded,alvcoli were collapsed,and fibrosis was obvious.These changes were less serious in Xuebijing group.④ Content of HYP (μg/g):contents of HYP in lung tissues on the 3rd day in PQ group and Xuebijing group were significantly higher than those in control group (743.3 ± 50.2,718.1 ± 34.0 vs.665.8± 6.6,both P＜0.05),it then increased gradually,but the contents of HYP in Xuebijing group were significantly lower on the 7th day and 14th day than those in PQ group (790.5 ± 23.8 vs.876.7 ± 42.0,812.9 ± 72.3 vs.931.3 ± 33.0,both P＜0.05).⑤ Expression of Mfn2:the expression of Mfn2 in control group was relatively lower.The expression of Mfn2 in PQ group was increased gradually under stress,but its rate was low.The expression of Mfn2 (A value) in Xuebijing group was significantly higher than that in PQ group on the 1st day (0.731 ±0.035 vs.0.618 ±0.029,P＜0.05),and it was elevated steadily,reaching the peak on the 7th day (0.732 ± 0.037 vs.0.669 ± 0.034,P＜0.05),but it was lower than that of PQ group on the 14th day (0.708 ± 0.034 vs.0.765 ± 0.041,P＜0.05).Conclusions Xuebijing reduces lung inflammatory reaction and pulmonary fibrosis as a result of PQ poisoning.The mechanism is that Xnebijing regulates and increases expression of Mfn2 in lung tissue.

Objective To study the effect of PcG member NSPc1 on proliferation of HeLa cells.Methods Using bioinfomatic analysis to design the siRNA sequence to knockdown NSPc1.Detecting the expression level of NSPc1 in HeLa cell line using semi-quantitative RT-PCR,Real-time PCR and Western blot after transfection of the designed siRNA.Transient transfecting pSUPER-NSPc1 into Hela cells and performing BrdU incorporation assay.Establishing NSPc1 stably knockdown cell line,comparing proliferation abilities with the control cells.Results(1)The designed siRNA did efficiently knockdown the expression of NSPc1;(2)Transient knockdown of NSPc1 could repress BrdU incorporation;(3) The established NSPc1-knockdown cell lines had a significantly lower proliferation rate than that of control cells.Conclusion The expression of NSPc1 is necessary for the normal proliferation of HeLa cells.The NSPc1 stably knockdown cell pool is a useful model for further study of pathway related to NSPc1.

Objective To compare the clinical effects of proximal femoral nail antirotation (PFNA) and dynamic hip screw (DHS) in the treatment of intertrochanteric fractures in elderly patients.And provide a reasonable basis for clinical treatment of elderly patients with intertrochanteric fractures.Methods The clinical and follow-up records of 58 elderly patients with intertrochanteric fractures treated by PFNA and DHS were retrospectively reviewed.For times of operation,blood loss in operation,length of incision,incidents of complication and Harris's hip functional standard score were compared and analyzed.Results PFNA group operation time[(59.61 ± 8.27)min],amount of bleeding[(234.51 ± 38.80)ml] were both better than the DHS group of[(83.54 ± 11.12)min and (446.57 ±54.01) ml] respectively.There were significant differences (t =9.80,18.10,all P ＜ 0.05).The satisfactory rate was 92.8％ in PFNA group,higher than the DHS group's 83.3 ％ (x2 =6.18,P ＜ 0.05).There was significant difference between two groups in the incident of complication,DHS group was higher.Conclusion PFNA internal fixation with a minimally invasive,fixed solid and recovery fast,is the better internal fixation in the treatment of elderly intertrochanteric fracture patients.

for the detection of their virulent abilities simultaneously.

Objective To investigate the correlation between the glial fibrillary acidic protein(GFAP),neuron-specific enolase(NSE),synaptoghysin (SYN),neurite outgrowth inhibitor-A(Nogo-A)expression and neurological outcome in tissue surrounding the infarct during the recovery after cerebral ischemia-reperfusion injury in rats.Methods A 2-hour middle cerebral artery occlusion(MCAO)and reperfusion model in rats was induced by the intraluminal suture method.The modified neurological severity score(mNSS)was performed at day 28,35,42,and 49.Immunohistochemistry was used to detect the expressions of GFAP,NSE,SYN,and Noga-A in tissue surrounding the infarct.Results The mNSS score decreased gradually over time after cerdnal ischemia-reperfusion injury in rats.Except day 35(5.11±0.737)vs.day 42 (4.54±0.519),and day 42 vs.day 49(4.29±0.488),there were significant differences at all other time points(all P<0.05).The numbers of GFAP positive cells deergased gradually form day 28 to day 49,in which,the numbers of GFAP positive cells at day 42(51.00±13.59)vs.day 49(44.38±11.94) were significantly less than those at day 28(69.00±15.10)(P<0.05).There were no significant differences in the numbers of NSE positive cells at all time points,but their integrated optical density(IOD)increased gradually.There were significant differences between day 28(6 218.57±1 864.25)and day 42(9 414.00±2 491.12) or day 49(12 522.50±3 106.99),and between day 35(7 343.40±1 533.35)and day 49(all P< 0.05).There were no significant differences at all other time points.The SYN express (IOD)increased gradually.and it was significantly lower at day 49(66 503.00±12 834.61)than that at day 28(43 905.14±13 208.59)(P<0.05).The numbers of Nogo-A positive cells decreased gradually,and they were significantly less at day 49(42.13±14.45) than those at day 28(59.57±15.25)(P<0.05).The GFAP expression was positively correlated with the mNSS scores(r=0.993,P=0.007).The NSE(r=-0.954,P=0.044)and SYN(r=-0.992,P=0.008) expression was negatively correlated with the mNSS scores.Conclusion The neurological outcome was associated with the downregulation of GFAP expression and the upregalation of NSE and SYN expression during the recovery after cerebral ischemia-reperfusion injury in rats.

OBJECTIVE:To establish a method for the simultaneous content determination of berberine hydrochloride,phello-dendrine hydrochloride and magnoflorine in Phellodendron amurense decoction piece,and to campare the contents of the 3 ingredi-ents in different grades of P. amurense decoction piece. METHODS:HPLC was performed on the column of Phenomenex Luna C18 with mobile phase of acetonitrile-0.05 mol/L KH2PO4 (gradient elution) at a flow rate of 1 ml/min,the detection wavelength was 280 nm,the column temperature was 30 ℃,and injection volume was 5 μl. RESULTS:The linear ranges were 0.387 0-7.740 μg for berberine hydrochloride(r=0.999 9),0.044 4-0.888 0 μg for phellodendrine hydrochloride(r=0.999 8)and 0.048 0-0.960 0 μg for magnoflorine(r=0.999 9);RSDs of precision, stability and reproducibility tests were lower than 3%, recoveries were 95.61%-103.22%(RSD=2.80%,n=6),96.18%-102.80%(RSD=1.84%,n=6) and 97.93%-102.78%(RSD=1.84%,n=6). CONCLUSIONS:The method is simple and accurate,and can be used for the contents determination of berberine hydrochloride, phellodendrine hydrochloride and magnoflonine in P. amurense. The contents of berbenine hydrochloride and phellodendrine hydro-chloride in the first-grade decation piece are higher than those in the second-grade decoction piece,and the content of magnoflorine in both decoction pieces shows no discernible differences.

Objective To investigate the effects of chronic hepatitis B virus (HBV) infection on human hepatic cytochrome P450 2C9 (CYP2C9).Methods Liver tissue samples and blood samples were obtained from 10 patients with chronic HBV infeetion and 10 healthy controls.CYP2C9 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.The activity of CYP2C9 was detected utilizing high performance liquid chromatography (HPLC).The expressions of CYP2C9 mRNA and protein were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blotting.The data were analyzed by t test.Results All the liver samples showed CYP2C9 wild-type (*1*1),while CYP2C9 (*2) and CYP2C9 (*3) were not detected.The maximum velocity (Vmax) of CYP2C9 in patients chronic HBV infection and healthy controls were (263.5±66.4) μmol/L and(284.6±85.9) μmol/L,respectively (t=0.614,P=0.5471).The expression of CYP2C9 mRNA in patients with chronic HBV infection (0.39±0.28) was significantly lower than that of healthy controls (0.65±0.13) (t=2.628,P=0.0171).Accordingly,the protein expression in patients with chronic HBV infection (0.26±0.13) was lower than that of healthy controls (0.60±0.19) (t=4.688,P=0.000 2).Conclusion The expressions of CYP2C9 mRNA and protein are decreased in chronic HBV infection which may down-regulate the enzyme activity.

Objective To explore the Monte Carlo calculation methods for the absolute dose calibration and output factor of 6 MV flattening-filter ( FF) and flattening-filter free ( FFF) photon beams based on TrueBeam accelerator .Methods The BEAMnrc code was used to model the LINAC head of FF and FFF modes.The BEAM_up covers the components from the target to the monitor chamber , and BEAM_down includes the structures beneath the chamber , the dose deposit to the monitor chamber contributed by the incidence electrons and scattered particles from the secondary collimators were calculated respectively .The incidence electron-induced dose at certain depths on the central axis were simulated by means of the DOSXYZnrc code .By means of dose calibration equation , the calibration factor for the standard field (10 cm ×10 cm) and the output factors for various fields (1 cm ×1 cm-40 cm ×40 cm) were computed respectively .Results For the 6 MV FF and FFF beams under the standard 10 cm ×10 cm field, 1 MU equals to 7.747 ×1013 ±3.099 ×1011 and 3.248 ×1013 ±1.624 ×1011 electrons to the target , respectively , which deposited 21.53 and 35.01 cGy to the monitor chamber of the virtual accelerator respectively .The difference between the simulated and calculated output factors were 0.72%±1.4%and 0.56%±0.78%for FF and FFF , respectively .Conclusions The model generated and measured output factors agree well , indicating the good accuracy of the dose calculation by the model , which would provides basis for further clinical dosimetric studies .