OBJECTIVESTo study the phenomena of hepatitis B virus (HBV) integration into the tissues of hilar cholangiocarcinoma (HCCA) and to identify the integration sites in the host genome.

METHODSTen fresh HCCA samples were collected from the tissues by surgical ablation, 1 normal hilar bile duct sample selected as control. Cellular DNA were extracted by Wizard SV Genomic DNA Purification System. PCR-derived assay (HBV-Alu-PCR) was employed to amplify the viral-host junctions which contain the HBV sequence and the adjacent cellular flanking sequences. The PCR products were purified and subjected to sequencing by ABI-3730XL Auto DNA Analyzer. The sequence analysis of viral-host junctions was performed by DNASIS MAX 3.0 bioinformatics software. The insertion sites between viral and cellular sequences were identified through homology comparison using NCBI BLAST and MapViewer search.

RESULTSIn 10 HCCA samples, 5 were demonstrated to have HBV integration fragments with total 6 inserted sites identified. Sequence analysis from viral-host junction showed that HBV X gene inserted into host genome at random distribution with truncated fragments. HBV integration recurrently targeted the unknown region in upstream of CXXC finger protein-1 (CpG-binding protein) gene (4 cases). p53 tumor suppressor gene was also found at the integration site.

CONCLUSIONSThere is high integration rate of HBV DNA into cellular genome of HCCA. HBV integration is found frequently into or close to cancer-related genes. The findings demonstrate that HBV infection might have association with the pathogenesis of HCCA.

Aged ; Base Sequence ; Bile Duct Neoplasms ; genetics ; virology ; Cholangiocarcinoma ; genetics ; virology ; DNA, Viral ; genetics ; Female ; Hepatitis B ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Virus Integration

OBJECTIVETo study the effect of Hepatitis B virus X (HBx) gene transfection on expression of human telomerase reverse transcriptase (hTERT) mRNA in human bile duct carcinoma cell lines QBC939 and to elucidate the significance of cis-activation of hTERT mRNA by HBx gene on the carcinogenesis of bile duct.

METHODSQBC939 were cultured in vitro and co-transfected with eukaryotic expression vector containing the HBx coding region and cloning vector containing enhanced green fluorescent protein (EGFP) coding sequence using liposome-mediated gene transduction technique. Thirty six hours after transfection, EGFP expression, the indicator of successful transfection in cells, was determined. Flow cytometry was applied to determine the transfection efficiency. Cells were harvested and total RNA was extracted with TRI(ZOL) Reagent. The expression of hTERT mRNA in QBC939 was assayed by Reverse Transcription Polymerase Chain Reaction. The expression of HBx protein in QBC939 was detected by immunocytochemistry staining and western blotting.

RESULTSThe transfection efficiency was 29.6% for both HBx expression vector and vector control group. The expression of hTERT mRNA was significantly increased when transfected with HBx expression vector than that transfected with OPTI-MEM medium and vector only. The expression of HBx protein could only be found in the cells when transfected with HBx expression vector by immunocytochemistry staining and western blotting.

CONCLUSIONHBx gene transfection may up-regulate the transcriptional expression of hTERT mRNA in bile duct carcinoma cells. The cis-activation of hTERT gene by HBx gene is primary mechanism for carcinogenesis of biliary epithelia after HBV infection.

Bile Duct Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; metabolism ; Trans-Activators ; genetics ; Transfection

OBJECTIVETo detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.

METHODSThe expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.

RESULTSThe positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.

CONCLUSIONhTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Telomerase ; analysis ; genetics

OBJECTIVETo explore causes of gluteal muscle contracture induced anterior knee pain and curative effect of arthroscopic release.

METHODSFrom March 2002 to August 2013,36 patients with gluteal muscle contracture induced anterior knee pain were treated, including 15 males, 21 females, aged from 9 to 40 years old with an average (18.7±7.2) years old; the courses of diseases ranged from 4 to 30 years. The clinical manifestations involved limited to symmelia, positive Ober sign, buttocks touch contracture belts, knee and patella slide to lateral when doing squat activities. All patients were performed gluteal muscle contracture release under arthroscopic. Postoperative complications were observed, Kujala scoring before and after operation was used for compare curative effect.

RESULTSAll patients were followed up with an average of 29 months. The incision were healed well, and no complications were occurred. Postoperative Kujala score were improved more than preoperative.

CONCLUSIONGluteal muscle contracture release could alleviate hypertension of lateral patella, and palys an important role in preventing patellofemoral arthritis.

Adolescent ; Adult ; Buttocks ; Child ; Contracture ; physiopathology ; Female ; Humans ; Knee ; Male ; Pain ; etiology

OBJECTIVETo evaluate the clinical outcome of arthroscopic repair method of meniscus injury with Fast-fix under local anesthesia.

METHODSFrom October 2005 to September 2012,106 patients with meniscus injuries admitted into our - hospital were studied, including 74 males and 32 females, ranging in age from 13 to 71 years old, averaged 27.6 years old. The duration of the disease ranged from 15 days to 5 years. The main clinical manifestations included knee joint pain after exercise, joint locking, pressing pain of knee joint and positive McMurray signs. The MRI showed meniscus tear or degeneration. Arthroscopic repairing surgeries were performed with Fast-fix under local anesthesia. Each patient was assessed with VAS pain evaluation and Lysholm knee-joint score system before and after operation. All the patients were followed up more than 1 year.

RESULTSOne hundred and 2 patients were followed up by recording subjective symptoms, clinical examinations and questions naires for an average of 2.6 years (ranged, 1.1 to 8 years), and 4 patients lost follow-up. All the 102 patients had no anesthetic complications. Ninety-six patients had normal subjective symptom and clinical examinations. Four patients had a mild ache with activities,2 patients had moderate pain after activities with joint space pressing pain. VAS pain evaluation and Lysholm knee-joint score after operation both were much better than that before operation.

CONCLUSIONonclusion: Local anesthesia can provide nice circumstances for surgeries. Arthroscopic repair using Fast-fix is an idea method for meniscus injury, especially for the posterior horn tear of medial meniscus, which is simple and convenient with less complications, and satisfactory results.

Adolescent ; Adult ; Aged ; Anesthesia, Local ; methods ; Arthroscopy ; methods ; Female ; Humans ; Knee Injuries ; surgery ; Magnetic Resonance Imaging ; Male ; Menisci, Tibial ; surgery ; Middle Aged ; Tibial Meniscus Injuries ; Visual Analog Scale ; Young Adult

OBJECTIVETo evaluate the application value of percutaneous endoscopic gastrostomy (PEG) for home nutrition support in patients with stroke and post-traumatic brain.

METHODSWe retrospectively analyzed the clinical data of the 16 patients with stroke and post-traumatic brain, including cerebral infarction (n = 9), cerebral hemorrhage ( n = 5), subarachnoid hemorrhage (n = 1), and cerebral trauma (n = 1). All these patients underwent PEG in our hospital because they were not able to be orally fed.

RESULTSThe weight, triceps skinfold thickness, mid-upper arm muscle circumference, and serum albumin, hemoglobin, and lympholeukocyte cell counts 30, 60, and 120 days after hospital discharge were significantly higher than those at hospital discharge (P <0.05 or P <0.01). The NIHSS scores 30, 60, and 120 days after hospital discharge were 14.0 +/- 1.3, 14.0 +/- 1.1, and 3.0 +/- 1.2, respectively, which were significantly lower than 16.0 +/- 1.2 at hospital discharge (all P <0.05). Complications included gastric contents retention (n = 1), backstreaming (n = 1), aspirated pneumonia (n = 1), and intra-cushion syndrome (n = 1).

CONCLUSIONSPEG for home nutrition support is useful for the treatment of patients with stroke and post-traumatic brain with dysphagia and malnutrition after long-term coma. It can help to avoid the deterioration of nutritional status and improve the quality of life.

Adult ; Aged ; Aged, 80 and over ; Cerebral Hemorrhage ; physiopathology ; Cerebral Infarction ; physiopathology ; Enteral Nutrition ; methods ; Female ; Gastroscopy ; adverse effects ; methods ; Gastrostomy ; adverse effects ; methods ; Hemoglobins ; metabolism ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Serum Albumin ; metabolism ; Stroke ; physiopathology ; Subarachnoid Hemorrhage ; physiopathology

OBJECTIVETo explore the effectiveness of hamstring tendon graft embedding reconstruction of medial patellofemoral ligament for the treatment of recurrent dislocation of the patella.

METHODSFrom March 2008 to June 2013,67 patients with recurrent dislocation of patella were treated, including 28 males, 39 females with an average age of 22 years ranging from 10 to 42 years old. The clinical manifestations involved knee joint instability,"giving way", sense of patellar dislocation, anterior knee pain. All 67 patients underwent arthroscopic hamstring tendon graft embedding reconstruction of the medial patellofemoral ligament. The curative effect were evaluated by preoperative and postoperative Lysholm score and Q angle changes.

RESULTSAll cases were followed up after operation and the mean follow-up time was(27.5±13.4) months(4 to 69 months). Postoperative incision were healed well and no patellar dislocation or subluxation occurred. Lysholm score improved from preoperative 76.35±2.86 to 82.71±2.29 postoperatively; Q angle decreased from preoperative (18.75±2.33)° to postoperative(13.28±1.75)°.

CONCLUSIONThe method for the reconstruction of medial patellofemoral ligament can provide enough tension, patellar stability.imoroving the function of knee ioint.

Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Patellar Dislocation ; surgery ; Patellar Ligament ; surgery ; Patellofemoral Joint ; surgery ; Reconstructive Surgical Procedures ; methods ; Recurrence ; Tendons ; transplantation

OBJECTIVETo detect the expression of HBV X gene (HBx mRNA) in extrahepatic biliary tract carcinomas and the adjacent non-cancerous tissues, and to analyzed the relationship between HBV infection and incidence of biliary tract carcinomas, thereby to elucidate the possible role of HBx in the carcinogenesis of biliary tract.

METHODSThe plasmid pSPX46 was digested by appropriate restriction enzyme. HBx fragment was obtained through gel extraction kit. The digoxigenin-labeled DNA probes for HBx mRNA were prepared by a random prime technique. The expression of HBx mRNA was detected in formalin-fixed- paraffin-embedded specimens from 71 cases of biliary tract carcinomas and 39 specimens of non-cancerous tissues adjacent to cancer by in situ hybridization. The correlations between HBx mRNA expression and clinicopathological parameters were statistically analysed in 71 cases of biliary duct carcinomas.

RESULTSForty-three of 71 malignant specimens had detectable HBx mRNA expression with a positive rate being 61%. Only 7 of 39 specimens of non-cancerous tissues adjacent to cancer had weak HBx mRNA expression, with a positive rate being 18%, and all these positive signals were found in the hyperplastic biliary epithelium. No significant correlation was found between HBx mRNA expression and clinicopathological parameters, but a strong positive correlation was found between HBx mRNA and protein expression.

CONCLUSIONThere is a high frequency of HBx mRNA expression in extrahepatic biliary tract carcinomas. HBV infection and its gene integration might play a role to certain extent in the development of biliary tract carcinomas.

Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Bile Ducts, Extrahepatic ; pathology ; Biliary Tract Neoplasms ; complications ; virology ; Female ; Hepatitis B ; complications ; virology ; Hepatitis B virus ; genetics ; Humans ; In Situ Hybridization ; Male ; Middle Aged ; Molecular Sequence Data ; RNA, Messenger ; genetics ; Trans-Activators ; genetics

OBJECTIVETo evaluate the effects of arthroscopic debridement for acute gouty arthritis of the ankle.

METHODSForty-one patients with acute gouty arthritis of the ankle were treated under arthroscopy from January 2010 to June 2012. All the patients were male, age in ranging from 28 to 69 years with an average of 43 years. Eighteen patients were in the left ankles and 23 in the right ankles; 12 cases were firstly attack and 29 cases were recurrent attack. Course of disease was from 2 weeks to 30 months. The American Orthopedic Foot and Ankle Society (AOFAS) Ankle-Hindfoot Scale score was used to evaluate the clinical effects. Number of acute attacks of gouty arthritis were observed.

RESULTSAll the patients were followed up at least 12 months. The mean AOFAS Ankle-Hindfoot Scale score increased from 58.44 +/- 9.45 preoperatively to 86.15 +/- 7.36, 83.41 +/- 9.22, 84.10 +/- 8.22 postoperatively at 6, 12, months and the last follow-up respectively. Swelling of the ankle were improved significantly, pain was relieved and the mean number of acute attacks of gouty arthritis decreased significantly.

CONCLUSIONArthroscopy is helpful for the diagnosis of acute gouty arthritis of the ankle and improvement of clinical symptoms and ankle function.

Adult ; Aged ; Ankle Joint ; physiopathology ; surgery ; Arthritis, Gouty ; physiopathology ; surgery ; Arthroscopy ; Debridement ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

Humans ; Liver Diseases ; drug therapy