Objective:To investigate the specific interference of OX40 gene expression induced by RNAi technique in 293T cell lines transfected with rat OX40 gene.Methods:293T cells were transfected with recombined plasmid pEGFP-N1-GFP/OX40,and the positive cell clones were selected by fluorescence protein observation and RT-PCR.One specific dicer siRNA targeted to OX40 mRNA was designed and synthesized,which shared no homology with exons of known human gene.Quantitative real-time PCR was performed to measure the inhibitory rate of target gene expression by comparing OX40 mRNA concentrations before and after siRNA transfection.Results:10 nmol/L siRNA-OX40 elicited the highest level of gene silence in 293T cells which was transfected with siRNA after 48 h (68.3?8.7)%);The time of maximal inhibitory effect was at 48-72 h [(61.7?8.4)%,(39.6?5.6)%].Conclusion:The exogenous OX40 expression can be significantly inhibited by treatment with specific siRNA in a dose and time -dependent manner in 293T cells,which may provide a useful profile for further investigation of inhibition of OX40 protein,and a promising control approach for preventing immune reaction.

Objective To summarize the clinical experience of liver transplantation in the treatment of hepatocellular carcinoma(HCC).Methods From June 2004 to March 2007,70 consecutive HCC patients underwent liver transplantation in our hospital,including classic orthotopic liver transplantation in 41 cases,and piggyback liver transplantation in 29 cases.All data of patients were retrospectively analyzed.Results All liver transplantation were successfully conducted.The average warm ischemia duration was 4.5 minutes,and cold ischemia duration was 8 hours.There were 3 cases of postoperative deaths.Surgical complications were intra-abdominal hemorrhage in 2 cases,and biliary anastomotic stricture in 4 cases.Sixty-seven cases were followed up for 12-33(average 21) months,10 cases had recurrence of liver cancer after transplatation and 1 dead.Conclusions Liver transplatation can used on selected patients with HCC.Successful liver transplantation relies on good quality of liver graft,and idealized technique of vascular and bile duct reconstruction are key factors of liver transplantation.Proper postoperative management can effectively reduce the complications of operation.

Objective To observe the changes of blood sugar following use of recombinant human growth hormone (rhGH) in patients with severe burn. Methods A total of 140 patients with severe burn but with no history of diabetes were randomly and equally divided into Group A ( 18-44 years old) and Group B ( ≥45 years old) according to WHO criteria. Then, two groups were randomly divided Group B0 (treated with normal saline), 35 patients per group. Change of blood sugar and amount of insuline for control of blood sugar were observed and analyzed. Results Of 140 patients in four groups,128 patients accomplished the test, with no statistical difference upon general conditions between groups (P ＞0.05 ). Within four weeks after test, the amount of insuline used for control of blood sugar was (2908.0 ± 153.5) IU and (724.1 ± 31.1 ) IU respectively in Groups A1 and A0, (3725.9 ± 88.4) IU and (801.8 ±22.2) IU respectively in Groups B1 and BO. The results showed that the patients needing insulin to control blood sugar in Group B were more than that in Group A ( P ＜ 0.01 ). The time for blood sugar recovering to normal range in patients with high blood sugar with no use of rhGH in Group A1 and Group B1 was (5.21 ±0.64) d and (5.03 ± 1.01) d respectively (P＞0.05). Conclusions The blood sugar of patients with severe burn will rise after use of rhGH. The age of the patients is positively correlated with changes of blood sugar. Standard use of rhGH does not induce diabetes.

Objective To compare the air permeability,water absorption and water locking properties of two different foam dressings,thus to provide theoretical and experimental evidence to alternative optimization for acute and chronic wound.Methods Five Mepilex foam dressings(group 1) and PermaFoam Comfort dressings(group 2) each was selected.Simulated wound exudation was made by NaCl and CaCl·H2O.The water-absorbing rate of dressings at post immersion 24 h (PIH),the water-absorbing speed of dressings at post immersion 1,5,10,20 min,the diffusion diameter of exudation dripped on the surface of dressings for 5 min,the beaker filled with exudation was sealed tightly by dressing for 24 h,and the weight was gotten before and after 24 h.Statistical analysis was performed.Results (1) The water-absorbing rate:the group 1(616±19)% was significantly higher than (313±13)% of the group 2 (t=29.137,P<0.01);(2) The water-absorbing speed:the group 1 (119.68±2.59)g·s-1·m-2,(24.39±0.62)g·s-1·m-2,(12.33±0.29)g·s-1·m-2,(12.33±0.29)g·s-1·m-2 were significantly higher than those of the group 2[(65.85±4.37)g·s-1·m-2,(13.82±1.03)g·s-1·m-2,(7.16±0.41)g·s-1·m-2,(3.66±0.12)g·s-1·m-2,t=23.704,t=19.708,22.947,31.764,all P<0.01];(3) The water holding capacity:the group 1 (5.66±0.15)cm was significantly higher than (2.2±0.12)cm of the group 2,(t=39.089,P<0.01);(4) The air permeability:there was no statistically significant difference between the two groups(t=0.189,P>0.05).Conclusion The Mepilex foam dressing is more suitable for the early stage of acute wound with large exudation in short time,while the PermaFoam Comfort dressings is better for chronic wound or the later period of acute with less exudation in a relative slow seepage velocity.

Objective To investigate the effect of blockading OX40-OX40L co-stimulatory signaling on the survival time of liver allograft in rat.Methods siRNA-expression vectors were constructed to targeting OX40.3～5 minutes before DA to Lewis orthotopic liver transplantation was performed,5×109 pfu of targeting OX40 siRNA plasmid DNA were diluted in 5 ml of phosphate buffered saline(PBS)and inlected intravenously into recipient Lewis rat over a period of 10 seconds.Serum IL-2 and IFN-γ levels were assayed by ELISA,and mix lymphocyte response(MLR)were tested by 3H-thymidine.Results The survival time of recipients in siRNA treatment group(74.0±9.3)was significantly longer than that in control group[(7.3±0.5)days].In experiment group,the inflammatory cell infihration and liver tissue structure destruction were very slight.The concentration of serum IL-2 was much lower in siRNA treatment group[(46±8.4)pg/ml]than that in control group[(286.5±14.6)pg/ml].Meanwhile,the concentration of serum IFN-γ was much lower in siRNA treatment group [(202.7±14.6)pg/ml]than that in control group[(1682.7±87.9)pg/ml].Conclusion Administration of OX40-siRNA can blockade OX40-OX40L co-stimulatory signaling pathway.hence inhibit the rejection of liver allograft.

Objective To establish a human HepG2 cell growth model under the low oxygen environment induced by cobalt chloride in order to observe the impacts of human HepG2 cell proliferation,cellular cycles and apoptosis,namely cellular growth conditions,under the low oxygen environment induced by cobalt chloride with different concentrations and to study the HepG2 cell growth mechanism under the low oxygen environment induced by cobalt chloride.Methods The human HepG2 cells in the logarithmic phase were randomized grouping as control group and CoCl2 experimental group with different concentrations (50 μm/L,100 μm/L,150 μm/L and 200 μm/L).① HepG2 cell proliferation was tested by MTT assay to calculate cell's suppression rate and draw HepG2 cell growth inhibition curves.② The move ability of HepG2 cells was observed by scratch test.③ The cellular apoptosis and periodic changes were detected using the flow cytometer Annexin-V FITC/PI double-staining and PI single staining methods.④HepG2 cell's Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Bcl-2 protein expression were detected by Western Blot.Results ① The test results obtained via MTT assay showed that CoCl2 suppressed the human HepG2 cell proliferation within a certain amount of time and concentration and presented a time-dose dependent relation.② Scratch damage trial suggested that the cobalt chloride suppressed the HepG2 cell migration and wound repair capacity and presented a concentration dependent relation.③ Flow cytometer' s test results revealed that the apoptosis rates (％) in control group and experimental group with different concentrations (50 μm/L,100 μm/L,150 μm/L and 200 μm/L) were 3.42,7.74,13.07,20.56,28.53 and 44.45 (P ＜0.05),respectively.The apoptosis rate of the experimental group was significantly increased compared with the control group,as well as showing a concentration dependency.The results of cellular cycles revealed that the cobalt chloride significantly suppressed the HepG2 cell's periodic changes along with increases of concentration,as well as blocked the cell cycle staying in phase G1,thereby suppressing cell proliferation.④Western Blot test:Compared with the control group,the Bcl-2 protein expression was significantly decreased in the experimental group after treatment of cobalt chloride with different concentrations.Conclusion Within a certain range,CoC12-indiuced low oxygen environment can suppress the human HepG2 cell proliferation and healing migration capacity,induce apoptosis and present a time-dose dependent relation.The mechanism is likely associated with decreases of Bcl-2 protein expression.

ured with different techniques.Standardized application of somatropins will not induce diabetes.

OBJECTIVETo evaluate the down stream involvement of the bile duct in hepatolithiasis.

METHODSMechanical damage to bile duct epithelia and long standing cholangitis as result of hepatolithiasis play an important role in the carcinogenesis of bile duct epithelia and stricture of the intra- and extra-hepatic bile duct. Macromorphological and microscopic changes in bile duct mucosa of 100 consecutive patients with hepatolithiasis were investigated using intra- or post-operative cholangioscopy. Biopsy specimens of lesions obtained during cholangioscopy were studied with immunohistochemical staining and flow cytometry to determine proliferative activity and DNA content. Five cases of well-proven cholangiocarcinoma were simultaneously studied as controls.

RESULTSOf the 100 patients, those with chronic cholangitis accounted for 86% (86/100), proliferative lesions 11% (11/100), adenomatous polyps 1% (1/100), and adenocarcinoma 2% (2/100). The obvious mucosal lesion associated with hepatolithiasis was located down-stream of the bile duct, predominantly in the hilar region, e.g. orifices of the right/left hepatic duct and common hepatic duct (73% mucosa lesions in the hilar region). The intensity of cancer embryonic antigen stain and the proliferative cell nuclear antigen index increased with the development of bile duct lesions. Aneuploid DNA presented mainly in the high degree malignant adenocarcinomas (> 80% of cases).

CONCLUSIONSThe obvious mucosal lesions associated with hepatolithiasis were located down-stream of the bile duct, predominantly in the hilar region (73% of mucosal lesions). The proliferative activity of examined bile duct mucosa lesions increased with the development of pathological deterioration, which may contribute to the development of hilar bile duct stricture and hilar cholangiocarcinoma.

Adult ; Aged ; Bile Ducts ; pathology ; Carcinoembryonic Antigen ; analysis ; Cholangiocarcinoma ; etiology ; Humans ; Lithiasis ; complications ; pathology ; Liver Diseases ; complications ; pathology ; Middle Aged ; Proliferating Cell Nuclear Antigen ; analysis

Objective:To investigate the feasibility of universal newborn hearing screening in countryside in order to provide reliable evidence in launching this program all over the countryside of China. Method:Subjects were 12 638 infants who were born in 9 counties from Jan 2004 to Dec 2005. TEOAE was used for the fast hearing screening. Infants were screened on the 2-7 days after the birth. The re-screening was conducted in 4-6 weeks if failed in the initial screening,and follow-up were provided continually if they also failed in the re-screening. Result; Ten thouand eight hundred and forty-five of 12 638(85. 8%) were screened including 9 963(91. 9%) normal newborns and 882(8. 1%) newborns with high-risk. Seven thouand four hundred and fifty (68. 7%) newborns passed the initial screening, and 3 395 (31. 3%) people failed. One thouand seven hundred and ninty-three (14. 2%) infants were refused to be screened.Only 2 536 (74. 7%) were re-screened on time, and 859(25. 3%) did not receive re-screening. One hundred and twenty were failed in the re-screening or first screening, and 79 (65. 8%)of them received diagnostic assessment. Among the infants received diagnostic assessment, 6(7.6%)ca-ses were found to have profound hearing loss in both ears, 9(11. 4%)cases were found to be severe hearing loss(7 in both ears and 2 in single ear) , 11(13. 9%)cases were found to be moderate hearing loss (5 in both ear and 6 in single ear), 26 (32. 9%) were found to have slight hearing loss (11 in both ear and 15 in single ears), and 27 (34.2%) were normal. Fifty-two infants were diagnosed as hearing loss with a prevalence of congenital hearing loss(in binaural and monaural) of 0. 5%(52/10845)and a prevalence of bilateral hearing loss of 0. 3%(29/10845). A prevalence of congenital hearing loss was 0. 2% (22/9 963) in well infants and 3. 4% (30/882) in high risk infants. Among the 13 cases of children with severe and profound hearing loss in both ears children, 8(61. 5%)cases were fitted with hearing aids and 1 (7. 7%) case was implanted with cochlear implants. Conclusion:It is necessary and feasible to conduct hearing screening program in the rural area. However, the suitable model to perform the program in the countryside needs to be set up as soon as possible in order to get more poor infants to participate into the hearing screening program for free and increase the screening rate.

OBJECTIVE: To investigate the feasibility of universal newborn hearing screening in countryside in order to provide reliable evidence in launching this program all over the countryside of China. METHOD: Subjects were 12,638 infants who were born in 9 counties from Jan 2004 to Dec 2005. TEOAE was used for the fast hearing screening. Infants were screened on the 2-7 days after the birth. The re-screening was conducted in 4-6 weeks if failed in the initial screening, and follow-up were provided continually if they also failed in the re-screening. RESULT: Ten thousand eight hundred and forty-five of 12,638 (85.8%) were screened including 9,963 (91.9%) normal newborns and 882 (8.1%) newborns with high-risk. Seven thousand four hundred and fifty (68.7%) newborns passed the initial screening, and 3,395 (31.3%) people failed. One thousand seven hundred and ninety-three (14.2%) infants were refused to be screened. Only 2,536 (74.7%) were re-screened on time, and 859 (25.3%) did not receive re-screening. One hundred and twenty were failed in the re-screening or first screening, and 79 (65.8%) of them received diagnostic assessment. Among the infants received diagnostic assessment, 6 (7.6%) cases were found to have profound hearing loss in both ears, 9 (11.4%) cases were found to be severe hearing loss (7 in both ears and 2 in single ear), 11 (13.9%) cases were found to be moderate hearing loss (5 in both ear and 6 in single ear), 26 (32.9%) were found to have slight hearing loss (11 in both ear and 15 in single ears), and 27 (34.2%) were normal. Fifty-two infants were diagnosed as hearing loss with a prevalence of congenital hearing loss (in binaural and monaural) of 0.5% (52/10845) and a prevalence of bilateral hearing loss of 0.3% (29/10845). A prevalence of congenital hearing loss was 0.2% (22/9,963) in well infants and 3.4% (30/882) in high risk infants. Among the 13 cases of children with severe and profound hearing loss in both ears children, 8 (61.5%) cases were fitted with hearing aids and 1 (7.7%) case was implanted with cochlear implants. CONCLUSION It is necessary and feasible to conduct hearing screening program in the rural area. However, the suitable model to perform the program in the countryside needs to be set up as soon as possible in order to get more poor infants to participate into the hearing screening program for free and increase the screening rate.
China ; epidemiology ; Feasibility Studies ; Hearing Loss ; diagnosis ; epidemiology ; Hearing Tests ; Humans ; Infant, Newborn ; Neonatal Screening ; Otoacoustic Emissions, Spontaneous ; Prevalence ; Rural Population