Purpose:To investigate the variant expression of human belomerase reverse transcriptase (hTERT) and NF ?B p65 protein and their relationships in gastric carcinogenesis.Methods:A total of 41 primary gastric cancers, 15 cases dysplasia, 23 cases intestinal metaplasia and 10 cases normal gastric mucosa were taken into this study. Expression of hTERTmRNT, hTERT protein and p65 protein were determined by in situ hybridization and immunohistochemistry. Results:①Among normal mucosa intestinal metaplasia dysplasia carcinoma,p65 expression were 0?34.18%(8/23)?53.33%(8/15)?60.98%(25/41),hTERTmRNA 10.00%(1/10)?39.13%(9.23)?66.67%(10/15)?85.37%(35/41) and hTERT protein 0?30.43%(7/23)?60.00%(9/15)?78 05%(32/41),respectively. All the three parameters were significantly increased in dysplasia and carcinoma, compared to normal mucosa, while the expression level were also significantly higher in carcinoma than in intestinal metaplasia ( P

[Objective] To investigate the onset of steroid-induced necrosis of femoral head. [Methods] A retrospective study was carried out in 162 cases of kidney disease treated with steroid. Among them, 62 cases (Group A) was complicated with steroid-induced femoral head necrosis (FHN) and the others without the complication are in Group B. [Resuits] In 62 cases complicated with steroid-induced FHN, patients in the age of 20-30 accounted 93.5%; female accounted 54.8%; those with high weight index accounted 80.6% and patients complicated with blood-stasis syndrome accounted 90%; 51.6% suffered from lupus glomerulonephritis; 96.8% was given large dose of steroid (over 1 rag' kg~(-1)?d~(-1)) continuously over 2 months; 83.9% was given steroid in a total dosage of over 5000mg within one year; 54.9% has been given dexamethasone and 83.9% medicated intravenously; 67.7% has not given blood-activating stasis-removing herbal drugs (BSHD) combined with or without anticoagulation drugs (ACD). Ninety percent suffered from necrosis of bilateral hip joints and 67.7% from bilateral hip-joint disease; 68.8% suffered from FHN at the stage Ⅲ and Ⅳ. The above indexes differed from those without steroid-induced FHN. [Conclusion] The incidence of steroid -induced FHN in kidney disease is high in the female and in the cases of lupus glomerulonephritis: those who are in obesity, complicated with blood-stasis syndrome, given large-dose steroid (over 1mg?kg~(-1)?d~(-1)) continuously over 2 months or in a short term, or medicated with dexamethasone, have the high risk of steroid -induced FHN; bilateral hip-joint disease is in the majority. The combination of BSHD and ACD can decrease the incidence of steroid -induced FHN in kidney disease.

Objective To investigate the effect of tuftsin and its inhibitor on pancreas microcirculation in acute pancreatitis(AP). Methods Sprague Dawley (SD) rats were randomly divided into five groups. Murine AP model was produced by retrograde injection of 4% sodium taurocholate into pancreatic duct, tuftsin or its inhibitor was injected at a dosage of 75 ?g/kg. At the time of 0,3, 6, 12 h, pancreas was harvested for pathology of microthrombus. esults Microthrombus in control group was not different with that in tuftsin group; At the time of 3、6 and 12 h microthrombus in other 3 groups significantly increased than control group and tuftsin group; With time, microthrombus in AP group、AP+ tuftsin group and AP+inhibitor group increased steadily and statistically significant; Tuftsin inhibitor significantly decreased microthrombus at the time of 6、12 h.Conclusions In acute pancreatitis tuftsin deteriorated pancreas microcirculation, which could be partially reversed by the administration of tuftsin inhibitor.

Objective To establish a cell model expressing the Langerhans cell-specific C type lectin receptor Langerin in vitro.Methods The cDNA of Langerin was obtained by PCR and cloned into a eukauotic green fluorescent protein (EGFP) expression vector pEGFP-C 1 with EGFP located in the N terminal region of the Langerin gene.Then,the recombinant plasmid was transfected into a human embryonic kidney carcinoma cell line HEK293T.Subsequently,laser confocal microscopy was performed to observe the expression of EGFP-Langerin fusion protein,and flow cytometry to measure the expression of Langerin.Laser confocal microscopy was also conducted to visualize the recognition and endocytosis of dust mite antigen (nDer p 2) by Langerin.Results PCR and Western blot confirmed the successful transfection of HEK293T cells with the recombinant plasmid as well as the expression of Langerin in the transfected cells.As flow cytometry revealed,the expression level of Langerin in transfected HEK293T cells was increased by 43％ compared with untransfected cells.Laser confocal microscopy showed that green fluorescein-labeled Langerin was successfully expressed,and could bind to and endocytose the red fluorescein-labeled antigen nDer p 2.Conclusions The fusion protein EGFP-Langerin expressed in HEK293T cells showed the distribution characteristic of cell surface receptors,and could bind to and endocytose allergens.

Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells (BMECs) co-cultured with umbilical cord mesenchymal stem cells (UC-MSCs) in serum-free culture mediuum.Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact.In the direct contact culture groups, UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶1, 1∶1, 1∶2, 1∶3, 1∶4, 1∶5, and 1:10, respectively.In the indirect contact culture group, the supernatant of UC-MSCs was used as the conditioned medium to re-suspend BMECs.In the control groups, UC-MSCs and BMECs were cultured alone.The cell growth status in each group was observed at 0, 4, 8, 12, 24, 36, 48, 60, 72 h after culture, and cell proliferation was detected by cell counting kit-8 (CCK-8) assay.Results At 48 h, the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups (P<0.05).Meanwhile, the optical density in the direct contact group at a concentration ratio of 1∶2 reached the peak, which was extremely significantly higher compared with the control groups (P<0.01) and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group (P<0.05).Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs, and the co-culture by cell-to-cell contact has a better effect.The optimal concentration ratio of UC-MSCs to BMECs is 1∶2, and the optimal culture time is 48 h.

Objective To optimize extraction technology of Fufang-Fuqing-Xiji.Methods Orthogonal test combined with Plackett-Burman experimental design was adopted to study the influence factors of water extraction in Fufang-Fuqing-Xiji. The contents of chlorogenic acid, paeoniflorin and caffeic acid in Fufang-Fuqing-Xiji were determined by HPLC gradient elution method at the same time. A multi-index comprehensive score method was applied to analyze the data and to screen the optimum extraction technology. Results The best extraction conditions of Fufang-Fuqing-Xiji were as following: threaded 80 mesh sieve, soaked 2 h, decocted 3 times with 12 times the amount of water for 1h each time.Conclusions This optimized extraction technology was rational and stable, which could be used for the extraction ofFufang-Fuqing-Xiji.

Objectives:To detect telomerase activity in various tumor tissues by PCR-PAGE and PCR-MPH.Methods:We used telomerase PCR-MPH，an assay in which a digoxigenin-labeled telomerase repeat-specific probe was hybridized to the PCR product.After revelation with an anti-digoxin-AKP conjugates,the amount of hybridized probe was determined by optical reading.Telomerase activity was analyzed by PAGE.Results:Telomerase activity was determined by PCR-MPH accurately and specifically in various tumor tissues,its sensitivity was one hundred times higher than that of PCR-PAGE.Conclusion:PCR-MPH is ideal for analyzing telomerase activity in various tumor tissues.It is simple,easy and suitable for clinical application.

Melanin is a kind of biopolymer, composed of a complex quinine/indole-quinone derived mixture which is produced in melanocytes from tyrosine. More than 100 genes related to melanin deposition have been found in mouse. Slc24a5 (solute carrier family 24, member 5) is a novel gene first cloned in zebrafish. And it has been confirmed that the Slc24a5 is involved in controlling the melanin deposition in zebrafish. Here the full length of the chicken slc24a5 mRNA sequence, its expression profile and a discussion of its relationship to melanin deposition in chicken were reported. Chicken Slc254a5 has a CDS of 1 269bp, predicting a protein of 423 amino acids which is about 80 amino acids shorter than in human and mouse. It has 9 exons and 8 introns and spans more than 1lkb of genome sequence. The quantitative real-time PCR confirmed that the chicken Slc24a5 was highly expressed in the eye of White Leghorn and Beijing Fatty Chickens, and in the eye, skin and muscle of Silky. The relative expression in Silky eye is more than two times that of White Leghorn. The relative expression in Silky skin is about 70 times that of White Leghorn and about 15 times that of Beijing Fatty Chickens. The relative expression in Silky muscle is about 15 times that of White Leghorn and about 3 times that of Beijing Fatty Chickens. And the expression result is in accordance with the melanocyte and melanin deposition in these tissues.

Objective To investigate etiology and clinical features of acute pancreatitis (AP) in young patients. Methods In total, 423 patients with AP admitted to Renmin Hospital of Wuhan University, Wuhan, Hubei province during January 2006 to December 2009 were retrospectively analyzed, 55 cases aged 34 years or less as young group and another 55 cases aged more than 34 years as middle- and elderly-age group. Their etiology, clinical features and illness scores based on some standard criteria were assessed. Results Proportion of varied etiologies of AP in young group was different from that in middle- and elderly-age one (x2 = 17. 107, P = 0. 009) , mainly as biliary and idiopathic pancreatitis in young group and as biliary and alcoholic pancreatitis in middle- and elderly-age one. Incidence of diet-related and idiopathic pancreatitis were higher in young group than that in middle- and elderly-age one ( x2 =4. 853, P = 0. 028 and x2=4. 274, P = 0. 039 ). Scores of the acute physiology and chronic health evaluation II ( APACHE II ) , Ranson criteria and the bedside index for severity in acute pancreatitis ( BISAP) were all lower in young group than those in middle- and elderly-age one (t = 5. 381 , P = 0. 000; t = 4. 388 , P = 0. 000 and t=3. 083, P = 0. 003 ) . There was no statistically significant difference in scores of computed tomography severity index ( CTSI) between young group and middle- and elderly-age one (t = ±0. 750, P > 0. 05 ) . Recurrence rate of AP was lower in young group than that in middle- and elderly-age one ( x2 =4. 251, P = 0. 039). Cholecystitis was more complicated in young group, pregnancy was more complicated in young group (x2 =4. 151 , P =0. 042) , and elevated blood glucose was more complicated in middle- and elderly-age one (x2 = 13. 285 , P = 0. 000 ) . Conclusions Etiology of AP varies in young patients with more dietary factors, occurrence of systemic complications and risks for death are lower in young group than those in middle- and elderly-age one, and there was no statistically significant difference in local complications and severity of illness between the two groups.

Objective To explore the difference of the level of Th1/Th2 balance and change of CD4+ CD45R cell after treated by laparoseopy-assisted rectal cancer surgery and conventional open rectal cancer surgery, so to compare the effect of the two procedures on immunity function of the patients. Methods CD4+ IFN-γ+ T cell (Thl cell), CD4+ IL-4 + T cell( Th2 cell)and the ratio of Th1/Th2 were detected by flow cytometery ; The levels of IFN-γand IL-4 were measured by ELISA. The changes of CD4 + CD45RA+ T cell and CD4+ CD45RO+ T cell were detected in the different procedures so that the effect of the two procedures on eytoimmunity and immune balance were analyzed. Results Compared with baseline levels, the level of Th1 cells was decreased [ (4.51±1.52 ) %, (7.26 ±2.59) % vs. ( 12.06 ±1.82 ) % ] (P < 0.05 ) ; The level of Th2 cells was increased [ (6.70 ± 2.41 ) %, (6.70 ± 2.41 ) % vs. (4.47 ± 1.90) % ] ( P < 0.05 ) ; The level of IFN-γ was decreased [ (57.15± 23.64) ng/L, (72.70 ± 27. 31 ) ng/L vs. ( 123.47 ± 32. 52) ng/L ] ( P < 0. 01 ) ; The level of IL-4 was increased [(55.55±7.29)ng/L,(57.56 ±7.13)ng/L vs. [(41.87±5.83)ng/L] (P<0.05) on 2nd and 7th day after conventional open reeatal cancer surgery; The level of Thl, Th2, IFN-γ and IL-4 in the laparoscopy-assisted rectal cancer surgery was not different ( P > 0.05 ). At 7 day postoperatively, the level of the CD4+ CD45 RA+ T cells was in-creased (35.11 ±7.82) and the CD4+ CD45RO+T cells was decreased(61.21±9.13) compared with surgery be-fore(31.11±6.72) and (68.11 ±11.42) respectively (P< 0.05). Conclusions The laparoscopy-assisted rec-tal cancer surgery has less influence on immune balance, thus protect systemic cell immunity.