Objective To identify the isolated rat bone marrow derived mesenchymal stem cells(MSCs),and evaluate the efficiency of adenoviral vector expressing human urokinase type plasminogen activator(uPA) in transfection of rat MSCs and its effect on proliferation of MSCs. Methods MSCs were isolated and purified by pasted wall purification,and were identified by immunicytochemistry.The transfection efficiency of uPA was detected by fluorescent microscopy,the expression of uPA in MSCs was detected by Western blotting,and the proliferation of MSCs was evaluated by MTT. Results The harvested MSCs exhibited the typical appearance of MSCs,and it was revealed by immunohistochemistry that the expression of MSCs markers CD29 and CD90 was positive,while that of CD34 and CD45 was negative.A tendency of increase in expression of green fluorescent protein(GFP) was observed with increase of multiplicity of infection(MOI).After transfection with AduPA for 72 h,the transfection efficiency reached(94.0?1.5)% at MOI of 80,and positive GFP cells could still be observed even after 7 d.The transfected uPA had no effect on the proliferation of MSCs. Conclusion MSCs are favourable genetic vectors to express uPA,and can be used for treatment of liver fibrosis.

Objective The infection of hepatitis B virus(HBV),which is a hepatotropic DNA virus,can cause acute or chronic viral hepatitis and further may develop liver fibrosis,cirrhosis and even hepatocellular carcinoma. In recent years,with the establishment of HBV transfection cell and animal models,the anti -hepatitis B virus effect of ingredients extracted from many traditional Chinese medicine has been proved.Therefore,chemical composition extracted from the resourceful library of traditional Chinese medicine,with high efficiency and low toxicity,might be the direction for developing anti -hepatitis B virus medicine.

Restriction site amplification polymorphism (RSAP) markers were employed to access the genetic diversity and relationship of 120 lilyturf germplasms from different geographical origins. Sixteen RSAP primer pairs generated 326 polymorphic bands, of which 318 (97.55%) were polymorphic. The value of polymorphism information content (PIC) ranged from 0.87 to 0.95 with an average of 0.92. These results indicated there was abundant genetic diversity among samples. The results of data analysis on 20 population showed that the value of percentage of polymorphic locus (PPL), Nei's gene diversity (H) and Shannon's information index (I) were 19.94%-85.58%, 0.082 6-0.210 7, 0.120 6-0.328 1 respectively. The most abundant genetic diversity was found in the O. japonicus population from Zhejiang and the least in the Liriope minor population. The genetic distance among 20 population was 0.024 6-0.286 8, of which the minimum genetic distance was 0.024 6 between population I and population 13 while the maximum 0.286 8 between population 5 and population 15. Coefficient of genetic differentiation among natural populations was 0.115 3 (Gst). And the gene differentiation contributed to 43.07% of the total genetic variation among populations and to 56.93% within populations. The total gene flow (Nm) was 0.660 9. UPMGA clustering analysis was basically similar to of the principle coordinate analysis (PCA). The 120 samples were classified into four major groups, which were basically corresponded with the genetic relationships based on morphological traits. The results of UPMGA and PCA were also consistent with geographical origins.
Amplified Fragment Length Polymorphism Analysis ; China ; Genetic Variation ; Liriope Plant ; classification ; genetics ; Phylogeny ; Polymorphism, Restriction Fragment Length

Drought stress exerts a considerable effect on growth, physiology and secondary metabolisms of the medicinal plants. It could inhabit the growth of the medicinal plants but promote secretion of secondary metabolites. Other researches indicated that the medicinal plants could depend on the ABA signaling pathway and secreting osmotic substances to resist the drought stress and reduce the damage by it. The article concludes the changes in growth, physiology, secondary metabolisms and response mechanisms of medicinal plants to drought stress that provides a theoretical basis for exploring the relationship between medicinal plants and drought stress.
Abscisic Acid ; metabolism ; Droughts ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Signal Transduction ; Water ; metabolism

With inlet temperature, specific gravity, feeding speed as independent variables, the comprehensive evaluating indexes of content of schisandrin and arctiin as dependent variable, the experimental data were fitted to a second order polynomial equation. Based on establishing the mathematical relationship between the comprehensive evaluating indexes and respective variables, Box-Benhnken central composite test and response surface analysis method was employed to optimize the spray drying technology of Biqiu granules ethanol extract. The optimal drying parameter was as follows: the inlet temperature was 175 degrees C, the specific gravity was 1.10, feeding speed was 32 r x min(-1). Under these conditions, the comprehensive evaluating indexes of spraying dry processes was 92.68, which was close to the model prediction. The spraying dry technology of Biqiu granules ethanol extract optimized by response surface methodology was accurate and feasible, which provided theoretical experiment basis for the industrialization production.
Chemistry, Pharmaceutical ; methods ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ethanol

First with the qualified rate of granules as the evaluation index, significant influencing factors were firstly screened by Plackett-Burman design. Then, with the qualified rate and moisture content as the evaluation indexes, significant factors that affect one-step pelletization technology were further optimized by Box-Behnken design; experimental data were imitated by multiple regression and second-order polynomial equation; and response surface method was used for predictive analysis of optimal technology. The best conditions were as follows: inlet air temperature of 85 degrees C, sample introduction speed of 33 r x min(-1), density of concrete 1. 10. One-step pelletization technology of Biqiu granules by Plackett-Burman design and Box-Behnken response surface methodology was stable and feasible with good predictability, which provided reliable basis for the industrialized production of Biqiu granules.
Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Temperature

OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.

METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.

RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).

CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.

Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the postoperative complications and safety of pancreaticogastrostomy (PG) versus pancreaticojejunostomy (PJ) after pancreaticoduodenectomy (PD).Methods Medline,EMBASE,Science Direct,Springer link,CBM,Cnki,Wan fang and VIP database were retrieved by computer search between 1st January 2004 and 31st March 2014 to collect all the RCT articles on pancreaticojejunostomy versus pancreaticogastrostomy after pancreaticoduodenectomy.The quality of the included trials was studied by assessing the inclusive and exclusive criteria (the PRISMA statement) by 2 researchers independently,then the data were extracted and analyzed using the RevMan 5.2.10 software.Results A total of 6 prospective randomized controlled trials which involved 976 patents were included in the study.There were significant differences between PG and PJ in terms of postoperative pancreatic fistula (RR:0.51 ; 95 ％ CI:0.37-0.70 ; P ＜ 0.0001),intra-abdominal fluid collection (RR:0.55 ; 95 ％ CI:0.34-0.89; P =0.01),and postoperative biliary fistula (RR:0.14; 95％ CI:0.03-0.59; P =0.0008).There was no significant difference in postoperative complications,mortality,delayed gastric emptying,postoperative hemorrhage,reoperation and length of hospital stay (P ＞ 0.05).Conclusions Pancreaticogastrostomy after pancreaticoduodenectomy is superior to pancreaticojejunostomy in safety and practicability.However,large,multicenter prospective randomized controlled trials are still needed to confirm the findings of this meta-anlaysis.

Objective To study the method of gray-scale blood flow imaging and image processing in condition of high frequency ultrasound,and the implementation of the system.Methods On the base of research of scattered signals of red blood cells in high frequency ultrasound,20 MHz ultrasound mechanical and linear scanning probe was used to transmit a number of pulses on a scan line.Pulse-echo subtraction method was used to obtain the blood flow information.At the end,simulated blood vascular was used to conduct flow imaging,and the obtained images were analyzed.Results Experiment results showed that clear blood flow images were obtained using this system.The noise from perivascular tissue could be filtered and the signals from blood flow could be enhanced after image processing.Conclusions In the detection of superficial blood vessel,blood flow signals can be obtained even using single pulse emitting via high frequency ultrasound.The blood flow imaging system can be implemented after image processing.