The 5'-flanking region of the human ?-fetoprotein(AFP) gene contains transcriptional regulatory sequences(TRSs) which up-regulate AFP gene expression with cell-specific enhancer activity in hepatoma cells. A couple of primers(Primerl: 5'-TGCAAGCTTATGATTCCCAAATATC-3'; Primer 2: 5'-GTCGAATTCGTGGCCTGGA TAAAGCTGAGT-3') were designed for synthesis and purification according to the known sequences of 5'-flanking region. AFPTRSs of 416 base pairs were amplified from human chromosome DNA by PCR, The identification of the AFPTRSs was confirmed to be consistent with reported sequences. The AFPTRSs can be applied to regulating the specific expression of cytokines in hepatoma cells.

In this study, the RT-PCR method was employed to detect the expression of AFP in mRNA level in tissue samples form 52 patients suffered from hepatocellular carcinoma (HCC) . The results revealed that the positive rate of AFPmRNA was 76.9% in the HCC tumor tissues and 69.4% in the paratumortissues from the HCC patients with severe cirrhosis . Meanwhile, in HCC patients without cirrhosis, the positive rate reached 50% in tumor tissues, but no AFPmRNA expression was found in related paratumor tissues. The study suggested that the AFP protein was specially expressed by hepatoma cells and mutating hepatocytes. The relationships between AFPmRNA and tumoor size, capsule status and tumor metastasis were also demonstrated.

Objective To investigate the expression of nerve growth factor receptors, P75 neurotrophin receptor (P75~(NTR)) and tyrosine kinase A (TrkA), and to explore their role in the pathogenesis of psoriasis vulgaris. Methods In situ hydridization and immunohistochemical technique were used respectively to detect the expression of P75~(NTH) and TrkA mRNA and proteins in lesional and non-lesional skins of 38 patients with psoriasis vulgaris (21 cases in progressive stage, 17 cases in static stage), and in normal skins of 10 healthy controls. Results In the case of expression of P75~(NTR) and TrkA mRNA and proteins, patients had significantly higher level than the normal control (P

To clone the full length cDNA of the tumor rejection gene MAGE-1 from hepatocellular carcinoma(HCC) tissues. This MAGE-1 gene and the tumor rejection antigen encoded by it may be useful in subsequent studies aiming at exploring new strategies for the immunotherapy for HCC. Methods: The full length MAGE-1 cDNA was amplified by RT-PCR method using a pair of primers designed according to the encoding sequence of MAGE-1 gene. The PCR products were then digested by restriction endonucleases and inserted into the plasmid PUC19. After primary selection of the recombinants by endonuclease digestion, the sequences of the inserted gene fragments were confirmed by DNA sequence analysis. Results; Using the same pair of primers, we obtained three clones of different MAGE genes, which were a full length MAGE-1 gene, a 750 bp fragment of MAGE-3 gene and a gene highly homologous to MAGE-6 and MAGE-12 but not identical to any reported MAGE genes. Conclusion: These data suggested that some MAGE genes are expressed in heptocellular carcinoma probably including some unknown genes, which might introduce potential new targets for immune attacks.

AIM: To investigate the influence and mechanisms of unsaturated fatty acid on PAI-1 expression in HepG-2 cells. METHODS: PAI-1 activity and mRNA expression were determined by colorimetric assay and RT-PCR, respectively. Two types of CAT reporter gene plasmid containing different deletions in PAI-1 promoter were constructed and transfected into HepG-2 cells, respectively. The transcriptional activity of PAI-1 was demonstrated by the CAT's expression. RESULTS: The mRNA and activity of PAI-1 significantly increased in the groups of oleic acid and linoleic acid compared with the control. When co-transfected with PPAR?-pSG5, the level of PAI-1 transcription was significantly increased. In the HepG-2 cells transfected with NF-?B-like sequence-deletion-pCAT construct linoleic acid, the PAI-1 transcriptional activity increased, and no significant change was observed when transfected with VLDL/fatty acid response element-deletion-pCAT construct. CONCLUSIONS: Unsaturated fatty acids induce PAI-1 activity and mRNA expression in HepG-2 cells. PPAR? may be one of transcription factors playing a role in the regulation of PAI-1 gene expression. The VLDL/fatty acid response element in the PAI-1 promoter may play an important role in the regulation, but not the NF-?B-like sequence.

OBJECTIVETo investigate therapeutic effects of vacuum sealing drainage (VSD) in the treatment of soft tissue defect combined with tendon and bone exposure.

METHODSFrom October 2007 to February 2011, 397 patients (412 feet) with open ankle fracture and dislocation combined with soft tissue defected were treated by VSD. There were 301 males and 96 females with an average age of 36 years (ranging age from 20 to 73 years). According to AO classification, 74 feet were type I, 211 feet were type II, 108 feet were type III and 19 feet were type IV. The mean time from injury to operation was 5.6 h ( 2 to 12 h). The mean treatment time of was 10 months (4 to 19 months).

RESULTSOne hundred and forty-one patients were primarily healed, 97 patients were sutured at stage II. Split-thick skin grafting was performed at stage II was performed in 103 patients; free flap transplantation was performed in 25 patients. Three of the 34 patients with infection were removed steel plate; Eviscerate flap coverage wound was performed in 14 patients caused by the first metatarsal bone exposure; Toe amputation were performed in 22 cases caused by toes necrosis. Tarsometatarasl joints perforators' surgery was performed in 10 patients with forefeet necrosis. Thirty hundred and six patients were followed up from 3 to 20 months (averaged 10 months). The wounds healed well.

CONCLUSIONVSD for soft tissue defects caused by ankle injury is a simple and effective method, but can not replace debridement and transfer flap.

Adult ; Aged ; Ankle Fractures ; Debridement ; Drainage ; methods ; Female ; Humans ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Skin Transplantation ; Treatment Outcome ; Vacuum ; Young Adult

Objective To investigate the efficacy of different radical surgical procedures for the treatment of hilar cholangiocarcinoma.Methods The clinical data of 207 patients with hilar cholangiocarcinoma who were treated at the Southwest Hospital from June 2007 to June 2012 were retrospectively analyzed.Local resection or hemihepatectomy combined with caudate lobectomy was applied to patients with Bismuth type Ⅰ hilar cholangiocarcinoma; dumbbell type radical resection was applied to patients with Bismuth type Ⅱ hilar cholangiocarcinoma or some patients with type Ⅲ a,Ⅲ b and Ⅳ hilar cholangiocarcinoma; hemihepatectomy or extended hemihepatectomy combined with caudate lobectomy was applied to patients with Bismuth type Ⅲ a,Ⅲ b and Ⅳ hilar cholangiocarcinoma.The patients were followed up every 3 months postoperatively till December 2012.All data were analyzed using the chi-square test or Fisher exact probability test,the survival curve was drawn by Kaplan-Meier method,and the survival was analyzed using the Log-rank test.Results Of the 207 patients,124 received radical resection,including 14 received local resection,23 received dumbbell type resection,87 received lobectomy + caudate lobectomy,49 received palliative resection; 34 received biliary drainage.Four patients died perioperatively.The incidences of complications of dumbbell type radical resection,left hemihepatectomy + caudate lobectomy,right hemihepatectomy + caudate lobectomy were 21.7％ (5/23),46.6％ (27/58) and 48.3％ (14/29),respectively.The incidence of complications after dumbbell type radical resection was significantly lower than left hemihepatectomy + caudate lobectomy and right hemihepatectomy + caudate lobectomy (x2 =4.42,3.90,P ＜ 0.05).One hundred and seventy patients were followed up.The median survival time of the 112 patients who received radical radical resection was 26.5 months,and the 1-,3-,5-year survival rates were 75.9％ (85/112),42.9％ (24/56) and 28.9％ (11/38),respectively.The median survival time of the 38 patients who received palliative resection was 8.5 months,and the 1-,3-year survival rates were 31.6％ (12/38) and 0.The survival time of 20 patients who received biliary drainage was 4.0 months,and the l-year survival rate was 0.The survival rate of patients who received radical resection was significantly higher than those who received palliative resection (x2=65.32,P ＜ 0.05).There was a significant difference in the survival rate between patients who received surgical treatment and those who received biliary drainage (x2=99.97,P ＜ 0.05).Of the 112 patients who received radical resection,the median survival time of 10 patients who received local resection of tumor was 47.0 months,the 1-year survival rate was 10/10,and 4 patients survived at the end of the follow-up; the median survival time of 23 patients who received dumbbell type radical resection was 32.0 months,and the 1-,3-year survival rates were 95.7％ (22/23) and 7/15,and the survival time of 6 patients was longer than 5 years; the median survival time of 54 patients who received left hemihepatectomy or extended left hemihepatectomy + caudate lobectomy was 27.6 months,and the 1-,3-year survival rates were 42.1％ (24/57) and 38.7％ (12/32),and the survival time of 9 patients was longer than 5 years,3 patients survived at the end of the follow-up ; the median survival time of 25 patients who received right hemihepatectomy or extended right hemihepatectomy + caudate lobectomy was 28.3 months,and the 1-,3-year survival rates were 45.8％ (11/24) and 6/15,and the survival time of 6 patients was longer than 5 years,2 patients survived at the end of follow-up.The median survival time of 35 patients (patients with Bismuth type Ⅰ,Ⅱ hilar cholangiocarcinoma and Bismuth Ⅲ a and Ⅲ b hilar cholangiocarcinoma which did not invade the secondary bile duct) who received hemihepatectomy + caudate lobectomy was 32.0 months,and the 1-,3-,5-year survival rates were 91.4％ (32/35),45.8％ (11/24) and 5/16,which were not different from the survival rate of patients who received dumbbell type radical resection (x2 =0.17,P ＞ 0.05).The 5-year survival rate of patients with lymph node metastasis was 4/19,which was significantly lower than 30.4％ (7/23) of patients without lymph node metastasis (x2 =23.40,P ＜ 0.05).Conclusion Joint lobectomy and standardized lymph node dissection could help to improve the efficacy of surgical treatment for patients with hilar cholangiocarcinoma.

Adult ; Creatine Kinase, MB Form ; metabolism ; Drug Therapy, Combination ; Electrocardiography ; Female ; Heart ; drug effects ; physiopathology ; Humans ; Hypoglycemic Agents ; therapeutic use ; Insecticides ; poisoning ; Insulin ; therapeutic use ; Isoflavones ; therapeutic use ; L-Lactate Dehydrogenase ; metabolism ; Male ; Myocarditis ; chemically induced ; drug therapy ; Myocardium ; enzymology ; pathology ; Organophosphate Poisoning ; Treatment Outcome ; Vasodilator Agents ; therapeutic use

In protection against microbes, an organism recognizes the pathogen associated molecular pattern (PAMP) on microbes by pattern recognition receptor (PRR). Toll-like receptor is called innate immunity. A family of cell membrane receptor was found in recent years that can mediate innate immune responses through the activation of a series of immune-related genes. In phylogenesis, it is highly conservative. However, its functions are getting more diversified with the complication of the immune functions of organisms.
Animals ; Humans ; Immunity ; Membrane Glycoproteins ; immunology ; Phylogeny ; Receptors, Cell Surface ; immunology ; Toll-Like Receptors

Objective To observe the microstructural changes of olfactory mucosa in rat model with acute rhinosinusitis leading to olfactory dysfunction,and to provide foundation for further exploration of corresponding mechanism.Methods On the basis of prior successfully established rat model of acute rhinosinusitis through inoculation with Streptococcus pneumoniae and with the help of merocel strips,one hundred healthy SD rats were randomly divided into experimental group (80) and control group (20).After inoculation,every 20 rats in the experimental groups were sacrificed in first week,second week,third week and fourth week respectively; and all rats in the control group were sacrificed in first week after the inoculation.Before the rats were sacrificed,the method called “buffed food pellet test,BFPT” was adopted,which was advanced by professor Nathan,to measure the rats’ olfaction,and the time of every rat spending in finding out the food pellet was recorded and analyzed.BFPT showed that the rats in experimental group spent (402.9±9.3),(453.7 ±7.3),(351.9±8.9),(278.7 ±8.1) s respectively in searching the food pellet,which were more than the rats in the control group[ ( 178.3 ±6.6) s].Then the olfactory mueosa was collected under anatomic microscope from all the rats to make frozen section and detect the changes of mature olfactory receptor neurons (ORN) and olfactory ensheathing cells (OEC) by immunofluorescence technique.Results The reduction of ORN in various degrees could be detected in the tissue samples of olfactory mucosa among all the rats in experimental group,with a tendency to become thinner in the thickness of epithelial lamina during the inflammation developing course.This kind of pathology was most marked in the second week and it gradually developed into the stage showing the lesion being the feeblest in the forth week following the beginning of modeling.Although the number of olfactory ensheathing cells appeared reduction in the first week following the beginning of modeling as well,it came to increase from the second week before olfactory receptor neurons and almost completely recovered to normal in the fourth week.In addition,some olfactory ensheathing cells could be detected in the tissue samples of olfactory mucosa among all the rats in experimental group.Conclusions Both mature olfactory sensory neurons and olfactory ensheathing cells appeared to reduction when sinonasal mucosa taken place acute rhinosinusitis.But the number of olfactory ensheathing cells increased faster than olfactory sensory neurons.In addition,some olfactory ensheathing cells could be detected in the olfactory epithelium.