Objective To study the expression of CCR4 in gallbladder carcinoma tissues,and to determine its relationship with clinical pathological factors and its influence on prognosis of gallbladder cancer.Methods The expressions of CCR4 in gallbladder carcinoma tissues,and chronic cholecystitis gallbladder mucosal tissues were detected using immune histochemical methods and they were analyzed together with the clinical records.Survival analysis was used to compare the expressions of CCR4 between the positive group and the negative group.Multiple factors analysis was carried out using the Cox regression model.Correlation between the expressions of CCR4 in gallbladder tissues and the clinical pathologic factors was done using the Chi-square test.Results CCR4 is expressed tan-yellow in gallbladder cell cytoplasm and/or cell membrane.The expression of CCR4 protein in the gallbladder carcinoma was obviously higher than in the chronic cholecystitis gallbladder epithelial tissues (P ＜ 0.05).A high expression of CCR4 was not correlated with patients' age,gender,pathological classification,distant metastasis and nerve/lymphatic invasion factors.It was,however correlated with tumor lymph node metastasis (P ＜ 0.05) and histological grading (P ＜ 0.05).Kaplan Meier survival analysis showed the postoperative survival between the CCR4 positive group and the negative group were significant different (P ＜0.05).On multiple factors analysis,CCR4 expression level was an independent risk factor of survival of patients after gallbladder surgery.Conclusions The chemokine receptor CCR4 expressed in gallbladder carcinoma.Its level of expression correlated with lymph node metastasis and histological grading.It was an independent risk factor of survival in patients with gallbladder carcinoma after surgery.As it was associated with invasion and metastasis of gallbladder carcinoma,antitumor treatment targeting CCR4 is expected to become a novel treatment of gallbladder carcinoma.

Ohiective To establish an in vitro test system for testing CMV specific immune response. Methods Human embryonic lung fibroblasts(HELF)infected with CMV-AD169 and a mutant strain RV-TB40E-4, respectively, were stained with anti-HLA-A * 0201-PE and then measured for the expression of HLA-A * 0201. The infected HELF were incubated with individual pp65 peptide NLVPMVATV and cultured together with PBMC,then intracellular IFN-γ was tested by flow cytometry(FCM). Results The expression of HLA-A * 0201 was markedly suppressed on AD169-infected fibroblasts, while it was not affected on RV-TB40E-4-infected cells. PBMC showing no response to AD169-infected cells responded significantly to RV-TB40E-4-infected cells. The HELF infected with AD169 and loaded with external peptides induced a significantly stronger response which was the sum of the induced response ratios of the infected HELF and the uninfected HELF loaded with peptides. Conclusion CMV-AD169 downregulates the expression of MHC Ⅰ, while it does not decrease the capacity of cells to present external peptides. RV-TB40E-4,a CMV mutant in which the gene of US2-6/US11 has been deleted, does not affect MHC Ⅰ expression, and therefore represents a more suitable tool to explore CMV-specific immune response.

Objective To research the function of the vagus nerve center nucleus of solitary tract(NTS)in the influence of gastric pressure by acupuncture at "Zusanli" in rats with motility disturbances of the stomach.Methods Adopting electrophysiological methods,the extracellular discharge signals of neuronic action in NTS were recorded with glass microelectrode.The influence of the discharges of NTS neurons and gastric pressure responding to the acupuncture at "Zusanli" point were observed in normal status and the condition of Atropine or Metoclopramide intravenous injection.Results In the normal status,acupuncture at "Zusanli" activated the discharges of NTS neurons and promote the gastric pressure remarkably.After Atropine intravenous injection,the background discharges of NTS neurons decreased.There was no statistical difference in the discharges responding to the effect of acupuncture before and after Atropine administration.And the gastric motion reinforced obviously by acupuncture at "Zusanli" after Atropine administration.But after intravenous injection of Metoclopramide,the background discharges of NTS neurons increased.There was also no statistical difference in the discharges responding to the effect of acupuncture before and after Metoclopramide administration.Moreover,the function of promoting gastric motion by acupuncture at "Zusanli" got more obviously after Metoclopramide administration.Conclusion The function of adjusting gastric motion by acupuncture at "Zusanli" point has close relation to the activation of NTS neurons,and the increasing amplitude of the discharges of NTS neurons wasn't influenced by cholinergic receptor blocker Atropine and dopamine-receptor antagonist Metoclopramide.

Purpose: To investigate the influence of needling bilateral points Hegu (LI 4) before colonoscopy on the examined patients and the time of inserting colonoscope. Methods: 185 patients to receive colonoscopy were randomly divided into an acupuncture group (93 cases) and a control group (92 cases) in order of examination. Bilateral points Hegu (LI 4) were needled before colonoscopy in the acupuncture group. No preventive measures were taken in the control group.Pain grades during the period that colonoscope passes the sigmoid and transverse colons were observed and the time of inserting colonoscope was recorded. Results: One hundred and sixty-five patients underwent colonoscopic insertion successfully, which were taken as the effective cases. Of them, 84 cases were in the acupuncture group and 81 cases in the control group. Statistics indicated that pain grades during colonoscope's passing the sigmoid and transverse colons were significantly lower in the acupuncture group than in the control group (P＜ 0.01); the time of inserting colonoscope was significantly shorter in the acupuncture group than in the control group (P ＜ 0.05).Conclusion: Needling bilateral points Hegu(LI 4) before colonoscopy can effectively relieve the patients'discomfort and shorten the colonoscopic time during the examination.

Objective To explore a new peptide-based approach independent of HLA to generate antigen-specific CD+ CD8+T cells. Methods Peripheral blood mononuclear cells(PBMC) were stimula- ted for 6 h with IE-1 peptide pool. Then the activated IFN-γsecreting ceils were tested by immunomagnetic selection. And the selected cells were cultured with radio-inactivated PBMC in medium with 100 IU/ml IL-2 for 4 weeks. Results The generated T cell lines consisted of IE-1 specific CD4+ T (6.88%) and CD8+ T cells 92.99%, which demonstrated antigen-specific killing and cytokine secretion. Conclusion T ceils can be proliferated with this new procedure, and maintain its phenotype and antigen-specific function.

Objective A new method is researched based on ECG for human identification.Methods Three features of wave feature,wavelet feature,fusion feature were selected as the feature vectors for 35 normal subject ECGs,and correlation coefficient discriminate analysis was used for human identification. Results The other 40 sessions from 35 subjects were used for verifying the accuracy of each feature.They were 82.5%、87.5% and 95% respectively for wave feature,wavelet feature and fusion feature. ECG wavelet feature outperformed ECG wave feature. While ECG fusion feature performed better than either feature. Conclusion The verification experiment demonstrates that the technology of human identification based on ECG is feasible. ECG can be combined with other biometric features such as fingerprint for developing the multi-biometric identification system. The method proposed in this paper is simple,real-time and exact for human identification.

AIM: To analyze the capability of cytomegaIovirus (CMV)-infected human embryonic lung fibroblasts (HELFs) to induce immune response. METHODS: HELFs were infected with cytomegalovirus and stained with antibody against HLA-A2 molecular, the expression of HLA-A2 was detected by FCM. The infected HELFs were incubated with individual pp65 peptide NLVPMVATV. While the uninfected and unloaded infected HELFs served as control respectively. After PBMC was added to the differently treated HELFs and incubated, the immune response was measured with IFN-γ release as readout. RESULTS: The expression of HLA-A molecular on infected fibroblasts diminished markedly compared with that on the uninfected. The peptides expressed on the infected HELFs together with those pulsed externally induced a stronger response than the infected HELFs alone. CONCLUSION: Although CMV can downregulate the expression of MHC Ⅰ on the infected cells, it can not decrease the capacity of cells to present peptides loaded externally, and therefore still induce immune response to some extent.

Background and purpose:Postoperative sore throat (POST) is one of the common complaints of patients after radical thyroidectomy. Tracheal intubation is the main cause of POST. This study compared the effect of intubation with visual endoscopy and general laryngoscope on POST in patients undergoing radical thyroidectomy. Methods:One hundred patients (18-60 years, ASAⅠ-Ⅱ) undergoing elective radical thyroidectomy were randomized into two groups:patients in group A (n=50) were intubated with visual endoscope while patients in group B (n=50) were intubated with general laryngoscope. Endotracheal tube cuffs pressure was maintained at 20mmHg in all patients. Visual analogue scale (VAS) and Bruggrmann comfort scale (BCS) were recorded at the time points of 1, 6 and 24 h after extubation. Results:Compared with group B, the incidence of POST in group A was signifcantly reduced (42%vs 64%, P=0.027). The VAS of group A was lower than that of group B (3.05±1.56 vs 4.25±1.30, 3.05±1.56 vs 4.01±1.98, 2.72±1.77 vs 3.31±1.12) (P<0.05). The BCS of group A was higher than that of group B (0.99±0.46 vs 0.69±0.30, 1.95±0.47 vs 1.51±0.58, 2.82±0.87 vs 2.31±0.72) (P<0.05). Conclusion:Using visual endoscopic intubation can reduce the incidence of the POST in patients undergoing radical thyroidectomy.

Aim Toresearchthemolecularmecha-nisms of adriamycin-induced cardiomyocyte apoptosis based on β-adrenoceptor signaling pathway during de-velopmentofheartfailure.Methods SeventymaleSD rats were assigned randomly into two groups:the con-trol group(CON,n=30)and the ADR-induced cardio-toxicity group (ADR,n =40 ).ADR was administered intraperitoneally in five equal injections (each contai-ning 3 mg·kg-1 )over a period of two weeks,with a total cumulative dose of 15 mg · kg-1 body weight. Age-matched rats injected with saline were used as controls.The general condition of all rats was observed, and transthoracic echocardiography was performed im-mediately following the final ADR injection,and then every other week.Serum and myocardial tissue were harvested at W2,W4 and W6 separately.The serum contents of brain natriuretic peptide(BNP)and cardiac troponin-T(cTn-T)were analyzed by euzymelinked im-munosorbent assay (ELISA ).The pathological change and apoptosis were determined by HE,Masson and ter-minal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL).The protein expressions ofβ1-AR,β2-AR,PKA and CaMK Ⅱ were detectedbyWesternblot.Results FollowingthefinalADRin-jection,cardiac systolic function and SV declined, which was accompanied by marked atrophy of the heart,low levels of cardiomyocyte fibrosis and apopto-sis,significantly increased serum BNP and cTn-T and decreased β1-AR,PKA and CaMK Ⅱ protein expres-sion.However,cardiac systolic function was improved with the extension of time but remained depressed as compared to CON group.The serum BNP and cTn-T concentration kept on rising.The gradual aggravation apoptosis and concomitant fibrosis in the ADR group heart were observed following ADR withdrawal.β1-AR protein expression was continuously down-regulated,β2-AR protein expression unchanged.Expression of PKA and CaMKⅡ proteins in hearts from ADR-injec-ted rats gradually increased.Conclusion β-AR/PKA/CaMKⅡ signaling pathway mediates cardiomyo-cyte apoptosis during the progress of ADR-induced car-diac dysfunction and pathological remodeling and apop-tosis.

Objective To study the effect of the combination of pathidine hydrochloride and anisodamine on the intraoperative effiacy of transhepatic biliary drainage and stent implantation done on the patient with advanced malignant obstructive jaundice.Methods 100 cases of percutaneous transhepatic catheterizde drainage (PTCD) and percutaneous transhepatic biliary stenting (PTBS) done for advanced malignant obstructive jaundice were divided into control group and experimental group,50 cases in each group.In experimental group,100 mg pethidine hydrochloride and 10 mg anisodamine were injected intramuscularly 30 min before operation,while in control group,no analgesic or antispasmodic were used before and during operation.The blood pressure and heart rate were observed,the incidence rate of biliary-cardiac reflex and operating time were recorded,while the scale of pain felt by patient was evaluated with visual analogue scale.Results Compared to the control group,the patients in experimental group have more stable vital signs,feel more comfortable,suffering shorter operative time,less incidence rate of biliary cardiac reflex and less pain by using pethidine hydrochloride and anisodamine.Conclusion Application of pathidine hydrochloride and anisodamine during the transhepatic biliary drainage and stent implantation done on the patient with advanced malignant obstructive jaundice may release pain,reduce operating time.It is safe,at lower cost and can be used as routine medicine before PTCD or PTBS.