Objective To investigate the expression of Muc1,Muc3A and Muc4 in cultured intrahepatic bile duct(IBD)tissues from different hepatic segments after cold preservation.Methods The IBD tissues of SD rats were obtained by collagen perfusion combining mechanical separation and then were divided into large and small IBD.The 2 parts of IBD were seeded in rat tail collagen gel and were cultured for 48 hours,then the IBD tissues from 10 rats were stored in UW solution at 4℃ for 1 hour(group I,n=5)and 12 hours(group Ⅱ,n=5),respectively,and the IBD tissues from the rest 5 rats were cultured in incubator at 37℃ for 24 hours (control group,n=5).The expressions of Muc1,Muc3A and Muc4 were detected by RT-PCR and Western blot.All data were analysed via ANOVA or LSD test.Results The expressions of Muc1,Muc3 A and Muc4 were detected both in large and small IBD tissues.The mRNA expressions of Muc1,Muc3A and Muc4 were decreased in large IBD as time passed by,which were 0.95±0.14,0.26±0.04 and 0.24±0.06 in group Ⅰ,0.18±0.03,0.14±0.04 and 0.22±0.07 in group Ⅱ,1.00±0.20,1.00±0.09 and 1.00±0.21 in control group,with significant difference among the 3 groups(F=8.8,57.1,10.8,P＜0.05).The mRNA expressions of Muc1 and Muc3A in group Ⅱ were significantly lower than in group Ⅰ(P＜0.05).The protein expressions of Muc1 and Mue3A in large IBD were also decreased as time passed by,which were 0.82±0.13,0.73±0.10 in group Ⅰ,0.56±0.11,0.33±0.04 in group Ⅱ,1.05±0.41,1.06±0.38 in control group,with significant difference among the 3 groups(F=3.9,12.6,P＜0.05).The protein expression of Muc1 of group Ⅱ was significantly lower than in control group(P＜0.05),and the protein expression of Muc4 in group Ⅱ was significantly lower than in group Ⅰ(P＜0.05).The mRNA expressions of Muc3A in small IBD were increased as time passed by,which were 0.15±0.04 in group Ⅰ,0.19±0.05 in group Ⅱ and 0.06±0.03 in control group.Conclusion Decreases of Muc1,Muc3A and Muc4 in IBD after long time cold preservation may weaken the selfprotection of biliary epithelium and case sever injury to bile duct.

Objective To compare the effects of different doses of amino acids and fat emulsions in parenteral nutrition on the incidence of complications and prognosis in very low birth weight infants (VLBWI). Methods The clinical data of 328 VLBWI who received nutrition support therapy for at least 5 days starting in 72 h after birth during January 2005 to December 2014 , were retrospectively analyzed. According to the dosage in parenteral nutrition, patients were divided into low-dose group and high-dose group. The incidence of complications and prognosis between two groups were compared. Results There were 204 cases in low-dose group and 124 cases in high-dose group. Compared with the low-dose group, the incidence of complications was lower in high-dose group during hospitalization and the incidence of intracranial hemorrhage was reduced most;the incidence of developmental retardation was lower at discharge;the overall incidence of metabolic complications of parenteral nutrition was higher, among which the incidence of high blood glucose, electrolyte disturbance and cholestasis were increased and the incidence of hypoglycemia was lower, and the differences were all statistically signiifcant (P??0 . 05 ). Conclutsions VLBWI can tolerate early aggressive parenteral nutrition which can reduce the incidence of extrauterine growth retardation and premature complications.

Objective To know the apolipoprotein E(APOE)genotypes frequency of Chinese old people in the rural area. Methods 2000 Chinese aged 65 years or older from four sites in China were enrolled in this study in 2004-2008.Two sites were from the Sichuan province in southwestern China,and another two sites were from the Shandong province in eastern China.The finger blood samples on filter paper were collected from all the investigated people in the end of the interview.The genotype for apolipoprotein E(gene symbol,APOE)was determined by eluting DNA from a dried blood spot,followed by HhaI digestion of amplified products.Results Through statistical analysis,APOE genotypes frequency of Chinese people aged 65 years or older in the rural area,as for ?/?2,?2/?3,?2/?4,?3/?3,?3/?4,?4/?4,they were 1.1%,13.55%,2.05%,68.80%,13.35% and 1.15% respectively.4 carriers and no 4 carriers were 16.55% and 83.45%.Conclusion The distribution of APOE 4 genotype is more widely in Chinese people aged 65 years or older in the rural area.

Objective:To learn the effect of personality in the course of cognitive decline.Methods:106 aged people from two districts were selected randomly.Arithmetic,digit span,picture filling,block patterns,digit symbol,visual attention span,spatial reasoning were used to learn the cognitive ability,“Kartaier's 16 personality factors test”were used to explore the personality of aged people.Results:16 personality factors test shows that old people are more deferential and self-assured.The relation between reasoning and arithmetic,rule-consciousness and arithmetic,abstractedness and digit span,reasoning and block design,dominance and digit symbol,sensitivity and special reasoning,apprehension and visual attention span,openness to change and visual attention span reach the statistical significance.Conclusion:Age is the most important reason to the cognitive impairment in aged people, education can prevent the cognitive impairment in a large degree.Personality can affect the cognitive ability although different personality factor affect different cognitive fields.

Objective To analyse the risk factors and prognosis of new on-set postoperative atrial fibrillation in different age and gender groups.Methods The study is a retrospective analysis in 516 patients from a single center.All cases were divided into 5 groups by ages as:＜ 40 years (n =61),40-49 years (n =97),50-59 years (n =115),60-75 years (n =140),and ＞ 75 years(n =103).We retrospectively analysed the difference in the occurrence-and recurrence-rate of POAF,risk factors,feature of coronary artery in different age and gender groups.Results There were no statistics differences in the same age group.The major risk factors were smoking,family history of coronary artery disease and hyperlipaemia in age ＜40 years group and 40 ～ 49 years group.while the major risk factors were hypertension and diabetes in the 60-75 years group and age ＞ 75 years group.The percentage of smoking patients decreased along with adding age.There was obviously statistical difference in the recurrence-rate of POAF across all groups during 24 follow-up months (P ＜ 0.05).The number of coronary lesion was increased with age.There were more ostial lesions in the 60-75 years group and age ＞ 75 years group.Conclusion There are different risk factors and characteristics among POAF patients in different age and gender groups.It is of great significance for us that comprehensive clinical intervention measures are taken to prevent recurrence of AF.

Objective To observe the effects of short hairpin RNA (shRNA) targeting Her2 on its gene expression when the shRNA was stably transfected into human ovarian cell lines, SKOV3 and SKOV3. ipl, which have different extent of malignancy and investigate the changes of the biological characters of the two cell lines after the stable transfection. Methods The plasmids expressing shRNA targeting Her2 gene were transfected into SKOV3 and SKOV3. ipl cells. The stably transfected cells were gained by antibiotic screening. The expression of Her2 before and after the transfection was detected by RT- PCR and western blot. The transwell experiment was used to observe the invasion ability of the cancer cells before and after the transfection, and the parent and the transfected cells were injected into nude mice to observe the tumor growth. Results After the stable transfection with Her2 shRNA, mRNA and protein levels of Her2 gene in SKOV3 and SKOV3. ipl cells were remarkably reduced. The expression of mRNA were (68.0±3. 1) %, (40. 8±2. 0) %, (99. 9±1.3) %, (42. 4±2. 5) %. The expression of protein were (72. 1±3.4) %, (36. 4±1.5) %, (98.2±1.7) %, (40. 7±2. 1) %. The invasion ability into basilar membrane of the transfected cells was greatly reduced compared with the parent cells. The invasion cell numbers were 7.6±1.1, 1.8±0. 8, 36. 2±9.7, 15.7±7. 2. The growth rate of the planted tumors was lower in transfected groups than that of the parent groups. Conclusions (1) The expression of Her2 gene in SKOV3 and SKOV3. ipl cells was remarkably reduced by RNA interference targeting Her2. (2) The biological characters of SKOV3 and SKOV3. ipl cells are changed when the expression of Her2 gene is reduced by RNA interference.

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

OBJECTIVETo investigate the role of endogenous hydrogen sulfide (H2S) in erectile dysfunction (ED) induced by androgen deficiency.

METHODSWe randomly divided 30 eight-week-old healthy male SD rats into six groups: 2-week control (A), 4-week control (B), 2-week castration (C), 4-week castration (D), 2-week castration + androgen replacement (E), and 4-week castration + androgen replacement (F), those in groups E and F subcutaneously injected with testosterone propionate (TP) at the physiological dose of 3 mg/kg per day after castration, while those in the other groups with isodose oil instead. At 2 and 4 weeks after operation, we determined the level of serum testosterone (T) , intracavernous pressure (ICP) , mean carotid arterial pressure (MAP) of the rats, measured the concentration of H2S in the plasma and corpus cavernosum tissue, and detected the expressions of cystathionine-P3-synthase (CBS) and cystathionine-gamma-lyase (CSE) by immunohistochemistry and Western blot.

RESULTSThe serum T level was significantly lower in group C ([0.63 +/- 0.15] nmol/L) than in A ( [ 16.55 +/- 4.17] nmol/L) and E ( [ 18.99 +/- 4.62] nmol/L) (P <0.05), as well as in group D ([0.70 +/-0.22] nmol/L) than in B ([15.44 +/-5.18] nmol/L) and F ([20.99 +/-6.41] nmol/L) (P <0. 05) , and so were ICP/MAP after 5 and 7 V electrical stimulation of the pelvic ganglia (P <0. 05) , H2 S concentration (P <0.05), and the expressions of CBS and CSE (P <0.05). The expressions of CBS and CSE proteins were also significantly decreased in group C as compared with D (P <0.05).

CONCLUSIONThe reduced expressions of CBS and CSE may inhibit the H2 S signaling pathway, which might be one of the mechanisms underlying androgen deficiency-induced ED in rats.

Androgens ; deficiency ; Animals ; Cystathionine beta-Synthase ; metabolism ; Cystathionine gamma-Lyase ; metabolism ; Erectile Dysfunction ; metabolism ; Hydrogen Sulfide ; metabolism ; Male ; Orchiectomy ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.
Apoptosis ; Cell Proliferation ; Dependovirus ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; Serpins ; metabolism ; Signal Transduction ; Transfection

The study is aimed to distinguish morphological characteristics of Dalbergiae Lignum collected from crude drug's markets and establish a identification methods and the quality standard for Dalbergiae Lignum. The macroscopic and microscopic features of Dalbergiae Lignum from crude drug's market were observed, analyzed and compared according to Hongmu specification issued by the People's Republic of China in 2000, and by the characteristics recorded in domestic monograph of Mucai Shibie (wood identification). The redwood of Dalbergiae Lignum cut into small pieces as medicinal material are dry heart wood of mahogany (trees from Dalbergia sp.), which characteristics of the small pieces as crude drug are different. There are differences in macroscopic and microscopic features about texture of wood and color, odor, taste, transverse section, radial section, tangential section. The results can provide basis for identification, application and improment of the quality standard of Dalbergiae Lignum as medicinal material.
China ; Dalbergia ; anatomy & histology ; chemistry ; classification ; Herbal Medicine ; economics ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Xylem ; anatomy & histology ; chemistry