Objective To investigate the effect of angiotenisn ⅡI (Ang Ⅱ) on RIN-m β-cell,and to explore the mechanism of β-cell function impairment caused by Ang Ⅱ.Methods RIN-m cells were cultured with various concentrations of AngⅡ (0.1,1,10,100 nmol/L).After incubation for 24 hours,the basal(3.3 mmol/L) and glucose-stimulated(16.7 mmol/L) insulin secretion(GSIS)were detected by radioimmunoassay,mRNA and protein expressions of uncoupling protein 2(UCP2)were determined by RT-PCR and Western blot,respectively.The intracellular ATP content was measured by luciferase bioluminescence.The mitochondrial membrane potential and cellular Ca~(2+) concentration were detected by flow cytometry.Results (1) Various concentrations of Ang Ⅱ had no significant influence on the basal insulin secrection of RIN-m cell(F=0.644,P = 0.634).Except for 0.1 nmol/L AngⅡ,the other concentrations of Ang Ⅱ markedly reduced GSIS of RIN-m cells(F= 118.528,P = 0.000).(2) Compared with the control group,Ang Ⅱ significantly increased mRNA and protein expression of UCP2(F= 1 370,P = 0.000;F=675.175,P = 0.000).(3)Except for 0.1 nmol/L Ang Ⅱ,the other concentrations of Ang Ⅱ significantly decreased the mitochondrial membrane potential,cellular ATP content,and cellular Ca2+ concentration of RIN-m cell(F=4.035,P=0.008;F=3.353,P = 0.013;F=5.867,P = 0.001).Conclusion Ang Ⅱ impairs GSIS of p-cell,the mechanism of impairment may be interpreted that Ang Ⅱ can increase the expression of UCP2,furthermore,it can reduce mitochondrial membrane potential,decrease the content of cellular ATP and the concentration of cellular Ca~(2+),can finally impair the function of β-cell.

Objective： To clone rat cardiac myosin α he avy chain cDNA fragment encoding aa736-960 and construct its recombinant retrov irus vector(RV). Methods: The 681 bp target gene was amplified f rom heart tissue of young rats with RT-PCR, fusion gene of huIL-2/myosin was c onstructed by splicing with huIL-2 cDNA using ligation methods and its RV was constructed. RT-PCR and immunohistochemical assay were used to iden tify the expression of myosin protein in transfected cell. Results: The determination of nucleotide sequence showed that the nucleotide and ami no acid sequence of gene clone was the same as reported, its openin g reading frame was correct, the digesting result of pLNC-huIL-2-myosin was i dentical with the predicted. NIH3T3 cell was transfected with recombinant RV, and G418-resistant NIH3T3 cell was established.RT-PCR analysis indicated tha t mRNA of pLNC-huIL-2-myosin was present in cell transfected with RV. The im munohistochemical assay also showed that the myosin protein expression was highe r in the cell transfected with constructed RV. Conclusion: Rat cardiac myosin α heavy chain cDNA has been cloned and its RV has also been cons tructed and expressed in NIH3T3 successfully, it will contribute to research of prevention and treatment of heart immune injury by cardiac myosin gene transfer to induce specific tolerance.

OBJECTIVETo study the occurrence, management and prognosis of fatal pulmonary embolism in patients underwent coronary intervention in our department.

METHODSeven patients had fatal pulmonary embolism after coronary intervention in six years, we analysis each patient by the occurrence, prognosis, management of the disease.

RESULTSDuring the last 6 years, 7 [five males, mean age (55.9 +/- 11.7) years, 5 after coronary angiography and 2 after percutaneous coronary intervention] patients developed fatal pulmonary embolism after PCI. All 7 patients presented respiratory and cardiac arrest within 24 hours post coronary intervention. Three patients died, one patient experienced brain death and another three patients survived and are alive without complication till now.

CONCLUSIONThe fatal pulmonary embolism is a scarce complication after coronary intervention with high acute mortality and satisfactory outcome for survivors.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Humans ; Male ; Middle Aged ; Prognosis ; Pulmonary Embolism ; diagnosis ; etiology ; Treatment Outcome

OBJECTIVETo analyze the CT image features of pneumonic-type lung cancer and to reduce misdiagnosis.

METHODSThe CT findings of 46 patients with pneumonic-type lung cancer were retrospectively reviewed, and CT image in the differential diagnosis of this special kind of disease was evaluated.

RESULTSAccording to the extent of lesion, these cases were divided into two groups: multi-lobe consolidation group and single lobe consolidation group. The lesions in the latter group located in the upper, middle or lower lobe, respectively. Twenty-nine cases had homogeneous consolidation lesion, 14 cases showed single or multiple cysts and cavities in the lesions, 3 cases exhibited localized low density in the lesion. Forty-one cases shown the sign of air bronchogram with presentation of narrow air bronchogram in 25 of those. Forty cases showed well or ill defined ground-glass opacitiy surrounding the consolidation lesion. Fifteen cases had multi-nodules or opacities distributed in centrilobular or centric bronchiolar location. Of the 30 patients who received contrast medium, 23 showed distinct enhancement, and 7 showed indistinct enhancement with a positive CT angiogram.

CONCLUSIONCT findings including lower lobe distribution, homogeneous consolidation, narrow air bronchogram, well defined ground-glass and CT angiogram are helpful in differentiating pneumonic-type lung cancer from various kinds of infection. However, most of CT manifestations of pneumonic-type lung cancer are not specific. Therefore, it's necessary to combine CT findings with other clinical data when making diagnosis.

Adenocarcinoma ; diagnostic imaging ; Adenocarcinoma, Bronchiolo-Alveolar ; diagnostic imaging ; Adenocarcinoma, Mucinous ; diagnostic imaging ; Adenocarcinoma, Papillary ; diagnostic imaging ; Adult ; Aged ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Follow-Up Studies ; Humans ; Lung ; diagnostic imaging ; Lung Neoplasms ; diagnostic imaging ; Male ; Middle Aged ; Pneumonia, Bacterial ; diagnostic imaging ; Radiographic Image Enhancement ; Tomography, X-Ray Computed ; methods ; Tuberculosis, Pulmonary ; diagnostic imaging

OBJECTIVETo study the treatment and therapeutic effects in the shearing-type and comminuted talar body fractures.

METHODSFrom October 1988 to September 2005,34 patients with shearing-type or comminuted talar body fractures were followed up. There were 19 males and 15 females ranged from 13 to 55 years (averaged 28.8 years). The disease course ranged from 3 to 14 days (averaged 6.0 days). Ten patients with a displacement of no more than 3 mm were treated with plastic cast. Eighteen patients were treated with open reduction and internal fixation, 6 patients were treated with joint fusion. The assessment of clinical efforts depend on patients' ache, active range of the joint and limp.

RESULTSThe mean follow up was 5.04 years (ranged from 3 to 19 years). All the patients were healed. The clinical outcomes were evaluated according to Hawkins evaluation score in which ache, active range of the joint and limp was respectively acssessed. There were 6 patients reached an excellent result, 9 good, 11 fair and 8 poor. Fifteen patients had osteonecrosis, 18 patients had traumatic arthritis of ankle joint, and 14 patients had traumatic arthritis of subtalar joint.

CONCLUSIONPatients whose displacement of fracture is not more than 3 mm should be treated with plastic cast. Operation and internal fixation should be performed in patients whose displacement of fracture is more than 3 mm after close reduction. Joint fusion should be performed in patients whose talar body fracture is comminuted severely and the surface of joint can not be repaired. The patients of talar body scissored fracture or comminuted fracture has bad prognosis.

Adolescent ; Adult ; Female ; Fracture Fixation, Internal ; Fractures, Closed ; surgery ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Talus ; injuries ; surgery ; Young Adult

OBJECTIVETo investigate the effects of local injection of insulin on the level of systemic blood glucose and granulation tissue formation of wound in patients with diabetic foot ulcer.

METHODSThirty-two patients with diabetic foot ulcer hospitalized in our wards from June 2009 to June 2010 were divided into insulin (I, n = 16) and control (C, n = 16) groups according to the random number table. For patients in I group, after debridement, one half of calculated dose of insulin diluted with equal amount of normal saline was injected diffusely into the base of the ulcer, and another half dose of insulin was subcutaneously injected into abdominal wall for 7 days, two times a day. For patients in C group, after debridement, primary insulin was subcutaneously injected into abdominal wall, 1 mL saline was subcutaneously injected into basal layer of ulcer for 7 days, two times a day. Before injection and 0.5, 1.0, 2.0, and 4.0 hours after injection (PIH), level of fasting blood glucose was determined. Before injection and on post injection day (PID) 3, 5, and 7, the growth of granulation tissue was assessed, and wound specimens were harvested for observation of CD34 expression and calculation of microvessel density (MVD). Data were processed with t test.

RESULTSThe levels of fasting blood glucose in both groups during observational time points ranged from 6.6 mmol/L to 12.8 mmol/L with a mean of (10.0 ± 2.2) mmol/L, and there was no statistical difference (with t values from 0.000 to 2.209, P values all above 0.05). Growth of granulation tissue in I group was more exuberant from PID 5, especially on PID 7 [(59.06 ± 1.58)%], which was significantly richer than that in C group [(23.61 ± 1.57)%, t = 17.420, P = 0.000]. New vessels were observed in I group from PID 3 as indicated by CD34 expression. There was no obvious difference in the number of MVD between I group and C group on PID 3 (t = 0.247, P > 0.05). The number of MVD per 200 times visual field in I group was respectively 8.34 ± 0.48, 11.22 ± 0.97 on PID 5 and 7, which was respectively higher than that in C group (4.42 ± 0.14, 5.44 ± 1.13, with t value respectively 16.568, 27.664, P values all below 0.01).

CONCLUSIONSLocal injection of insulin has a significant effect on systemic blood glucose in patients with diabetic foot ulcer, and it can promote the growth of granulation tissue and wound healing.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; metabolism ; Diabetic Foot ; drug therapy ; metabolism ; Female ; Humans ; Injections ; Insulin ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Wound Healing

OBJECTIVETo investigate the possible role of STAT3 and p38 in the carcinogenesis of sporadic colorectal tubular adenoma.

METHODSThe expression of STAT3 and p38 at protein level was studied in 107 sporadic colorectal tubular adenomas with different dysplasia (SCTA-D) or with cancerous changes (SCTA-Ca) by immunohistochemical staining method, meanwhile the expression of STAT3 at mRNA level was detected by in situ hybridization.

RESULTSImmunohistochemical staining results showed that the positive expression rate of STAT3 and p38 was 12.0%, 59.0%, 91.7% and 8.0%, 47.0%, 91.7% in normal colorectal mucosa (NCM), SCTA-D and SCTA-Ca, respectively, with a statistically significant difference of STAT3 and p38 expression among the SCTA-D, SCTA-Ca and NCM (P < 0.05). The expression of STAT3 and p38 was positively correlated with the degree of dysplasia from mild to severe SCTA-D (P < 0.05). In situ hybridization results showed that the positive expression rate of STAT3 at mRNA level in NCM, SCTA-D and SCTA-Ca was 8.00%, 51.8% and 100.0%, respectively, with a statistically significant difference among these either (P < 0.05). The positive expression of STAT3 at mRNA level was not only positively correlated with the degree of dysplasia (P < 0.05), but also with the expression of p38 (P < 0.05).

CONCLUSIONSTAT3 and p38 may be involved in the carcinogenesis of sporadic colorectal tubular adenoma.

Adenocarcinoma ; metabolism ; pathology ; Adenoma ; metabolism ; pathology ; Cell Transformation, Neoplastic ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Intestinal Mucosa ; metabolism ; Precancerous Conditions ; metabolism ; pathology ; RNA, Messenger ; metabolism ; STAT3 Transcription Factor ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo analysis the genetic mode of Rh DEL phenotype and RHD 1227A allele in Zhejiang Han population through family investigations.

METHODSRh DEL phenotypes were identified by a serologic adsorption-elution method. Two polymerase chain reaction-sequence specific prime (PCR-SSP) methods which detectED RHD 1227A allele and Rhesus hybrid box, respectively, and a nucleotide sequencing method focused on the exon 9 of RHD 1227A allele were employed to determine the zygosity of RHD allele.

RESULTSAll five probands with Rh DEL phenotype harbored a RHD 1227A allele and had a RHD allele deletion, and they were RHD 1227A/RHd heterozygote. One of the parent members was found to contain a RHD 1227A allele and a normal RHD allele in pedigree 1, 2 and 3, respectively. Thus, they were RHD 1227A/RHD heterozygotes and presented normal D positive phenotype. The son of proband No 1. inherited the RHD 1227A allele and presented a normal D positive phenotype due to a RHD 1227A/RHD heterozygote; The offsprings of proband No. 2, No. 4, and No. 5 did not inherit RHD 1227A allele and presented a normal D positive phenotype.

CONCLUSIONRHD 1227A allele is an important genetic marker of Rh DEL phenotype; RHD 1227A is recessive to normal RHD allele and dominant to RHd allele; RHD 1227A allele is an ancestral, but not a spontaneously mutated allele.

Alleles ; China ; Female ; Genotype ; Humans ; Male ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; methods ; Rh-Hr Blood-Group System ; genetics

BACKGROUNDCancer-testis antigen (CTA) is a family of the most noticeable tumor antigens which could be potential tumor markers for cancer diagnosis. In this research we aimed to investigate the expression of SSX-1 and NY-ESO-1 mRNA, two members of the CTA family, in tissue and peripheral blood of patients with hepatocellular carcinoma (HCC) to assess their feasibility for the immunotherapy and diagnosis of HCC and the association of their expression levels with diverse clinical indicators.

METHODSThirty-six north Chinese patients with HCC and 30 normal controls were enrolled in this study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and corresponding levels in peripheral blood of patients.

RESULTSThe positive rates of SSX-1 and NY-ESO-1 mRNA expression were 61.1% (22/36) and 11.1% (4/36), respectively, in cancer tissues; 38.9% (14/36) and 5.6% (2/36), respectively, in the corresponding peripheral blood samples. No positive expression of either SSX-1 or NY-ESO-1 mRNA was detected in the samples of cancer-adjacent tissues, cirrhotic tissues, normal liver tissue or the peripheral blood of control patients. No significant relationship was found between the expression of these two genes and clinical indicators such as age, gender, tumor size, extent of differentiation, serum a-fetoprotein (AFP) level or infection with hepatitis B virus (P > 0.05). The short term recurrence rate was 46.2% (6/13) in patients whose peripheral blood expressed SSX-1 mRNA, while the recurrence rate in patients with negative SSX-1 mRNA was 28.6% (4/14).

CONCLUSIONSSSX-1 and NY-ESO-1 antigens might be new potentially promising targets for antigen-specific immunotherapy for HCC. High specific expression of SSX-1 and NY-ESO-1 mRNA suggested that we could apply them as tumor markers. The short term recurrence rate was significantly higher in patients whose peripheral blood expressed SSX-1 mRNA, suggesting that SSX-1 mRNA could be used as indicator for recurrence, metastasis and prognosis of HCC.

Adolescent ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; genetics ; Neoplastic Cells, Circulating ; RNA, Messenger ; analysis ; blood ; Repressor Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the role and significance of Wnt/beta-catenin signaling pathway regulating GSK-3beta, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC).

METHODSThe HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against beta-catenin. Proteins were extracted and the expressions of beta-catenin, GSK-3beta, p-GSK-3beta, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection.

RESULTSbeta-catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P < 0.05). Interestingly, GSK-3beta and p-GSK-3beta expressions increased gradually at 72 and 96 h (tGSK-3beta= 4.98, tp-GSK-3beta= 29.83, P < 0.05) respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P > 0.05). Smad3 expression was increased at 72 h (t = 10.67, P < 0.05) and decreased to normal at 96 h (t = 1.26, P < 0.05), while TERT expression decreased at 72 h (t = 4.18, P is less than 0.05) and increased to normal at 96 h (t = 1.26, P > 0.05).

CONCLUSIONSWnt/beta-catenin signaling pathway is related to the expressions of GSK-3beta, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/beta-catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Small Interfering ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism