Objective To investigate the relationship between heart rate variability(HRV) and the carotid intima media thickness in type 2 diabetic patients.Methods One hundred thirty-two type 2 diabetic patients were categorized into groups by carotid intima-media thickness(IMT):group T1(36 cases,IMT

Acute coronary syndrome(ACS) belongs to “thoracic obstruction” and “heartache” in traditional Chinese medicine.Although many ancient physicians has discussed this disease,while due to the impact of “Yang Wei Yin Xian” in Zhang Zhong-jing's theory,the disease caused little attention by far.In the clinical practice,the author found that most ACS patients belong to heat syndrome.Based on TCM constitution theory,the author put forward the ACS was mainly caused by heat accumulation and blood stasis due to the changes of living environment,lifestyle,and diet,etc.According to traditional Chinese medicine theory,emotional disorder,spicy and hot food can burn body fluid,leading to blood stasis.Long term of blood stasis turns to heat,which further consumes heart blood and aggravates blood stasis.Therefore heat accumulation and blood stasis should be paid attention in the treatment of ACS.

Objective To establish a method for quantitative detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA ) in infected cells. Methods The transfected cell line HepG2.2.15 which can consistently produce Dane particles was maintained in DMEM containing 380 ?g/ml G418 and 10% fetal bovine serum. Cells in the exponential period were harvested from flasks, then intracellular HBV cccDNA was extracted from pellet containing 1?10~6 cells with mini plasmid extraction kit (QIAGEN).The extraction product was further purified by mung bean nuclease to remove HBV relaxed circular DNA possibly remained. HBV cccDNA was quantitatively detected by fluorescent PCR with selective primer set and Taqman MGB probe. Culture medium before exponential period, HBV DNA positive and negative serum samples from patients with chronic hepatitis B (mild) were amplified simultaneously to test the specificity of the fluorescent PCR method. Plasmids containing whole HBV genome were amplified with the same primer set and fluorescent probe to determine the sensitivity of the method. Results HBV cccDNA was detectable in HepG2.2.15, and the average quantity was 18 copies per cell approximately. No detectable fluorescent signal was observed when culture and serum samples were amplified. The detectable HBV cccDNA was as low as 10~3 copies per ml at least by this method. Conclusions This method is convenient, highly specific and highly sensitive. It can be utilized in the quantitative detection of intracellular HBV cccDNA as well as in the screening and evaluation of antiviral agents.

Amyloid beta-peptides ( Aβ) is the key pathological feature of Alzheimer’s Disease (AD). Various factors contrib-ute to the accumulations of Aβ in the brains of patients. Among them, blood-brain barrier ( BBB) plays a crucial role in trans-porting Aβ between the brain and the bloodstream while this transfer function is mediated by the receptor of Aβon BBB. The abnormality of Aβ transport and related receptor expression can be detected in the brains of patients with AD, resulting in an un-usual increase in Aβlevels unusually increased . This review e-laborates the structure and function of BBB, the transport of Aβand the expression and transport mechanism of the related recep-tor, as well as summarizes the possible clearance strategy of Aβacross the BBB.

OBJECTIVETo study whether the dose of controlling antihypertensive drug is reduced by transcutaneous electrical acupoint stimulation (TEAS) and the anesthetics, as well as the control of blood pressure (BP) and heart rate (HR) in endoscopic endonasal surgery with general anesthesia.

METHODSSixty patients for selective endoscopic endonasal surgery with general anesthetics and controlling antihypertension involved were selected and randomized into a TEAS group, a sham-TEAS group, 30 cases in each one. The electric pads were attached to bilateral Hegu (LI 4), Zusanli (ST 36), Sanyinjiao (SP 6) and Quchi (LI 11), stimulated with Hans-200 apparatus, 3 to 5 mA, 2 Hz/100 Hz in the TEAS group based on the patients' response to comfort. No electric stimulation was applied to the sham-TEAS group. The general anesthesia started after 30 min intervention and lasted till the end of surgery. The BP and HR were observed and recorded at the end of monitoring in operation room, 10 min after tranquilization (T0), 30 min after intervention (Tj, after induction~of general anestiesa (T2), 30 min after surgery start (T3), 60 min after surgery start (T4) and 30 min after extubation (T5). The doses of vecuronium bromide, propofol and nitroglycerin were recorded statistically in surgery, as well as the operative bleeding volume, the operative time, the resuscitation time and the visual analogue scale (VAS) score after resuscitation.

RESULTSCompared with that at T0, the mean arterial pressure (MAP) at T2, T3, T4 and T5 in the TEAS group and at T3 and T4 in the sham-TEAS group was all reduced, indicating the significant difference (all P < 0.01). MAP at T2 and T5 in the TEAS group was lower than that in the sham-TEAS group (both P < 0.01). Compared, with that at T5, except at T2 in the TEAS group (P<0. 05), HR was not different significantly at the rest time points (all P > 0.05). HR was different at T2 to Ts in the sham-TEAS group statistically (all P < 0.01). The doses of vecuronium bromide, propofol and nitroglycerin, the operative bleeding volume, the operative time, the resuscitation time and VAS after resuscitation were not different significantly between the two groups (all P > 0.05).

CONCLUSIONThe general anesthesia with TEAS and anesthetics involved for controlling antihypertension contributes to the control of BP and HR in the patients in endoscopic endonasal surgery. The impacts are not obvious on the doses of antihypertensive drug, the general anesthetics, the operative bleeding volume, the time of resuscitation and the postoperative analgesia.

Acupuncture Analgesia ; Acupuncture Points ; Adult ; Anesthetics, General ; administration & dosage ; Blood Pressure ; Electric Stimulation ; Endoscopy ; Female ; Heart Rate ; Humans ; Male ; Middle Aged ; Nose Diseases ; physiopathology ; surgery ; Young Adult

Objective:To evaluate the expression of vascular endothelial growth factor C(VEGF-C),Epstein-Barr virus(EBV) latent membrane protein 1(LMP1),cyclooxygenase-2 (COX-2) in nasopharyngeal carcinoma(NPC).Method:LMP1, COX-2 and VEGF-C were detects by immunohistochemical staining for 57 case NPC tissue.Result:The positives rates of LMP1,COX-2 and VEGF-C detected by immunohistochemical staining were 49.1%(28/57), 75.4%(43/57)and 59.6%(34/57), respectively. The expression of LMP1, COX-2 and VEGF-C were correlated to each1 other in NPC(P<0.05).Conclusion:LMP1 and COX-2 may induce expression of VEGF-C directly or LMP1 induce expression of VEGF-C by induce COX-2 expression, may contribute to lymph metastasis and develop NPC.

Objective To investigate the possibility of transfection of adipose-derived mesenchymal stem cells (ADMSCs) into the insulin-secreting cells in vitro,and assay the secretion of insulin of ADMSCs in high and low glucose environment.Methods The ADMSCs that untransfected were in the control group,the ADMSCs that contained PcDNA3.1 were in the vacant vector group,and the ADMSCs that contained PcDNA3.1-hINS were in the recombinant vector group.After transfection,the recombinant vector group were sub-divided into the 1,6,12,18 days groups.According to the concentrations of glucose,the recombinant vector 18 days group were divided into the high glucose group and low glucose group.Human insulin gene was amplified by RT-PCR,and the eukaryotic expression recombinant vector PcDNA3.1-hINS that contained the human insulin gene was constructed.The ADMSCs were obtained by digestion and centrifugation,and then underwent flow cytometry for identification.The transcription of insulin DNA was assayed by RT-PCR,and the levels of insulin were assayed by ELISA.Glucose test was done in the recombinant vector 18 days group.The measurement data was shown in the format of (x) ± s,the measurement data in multiple groups were compared by randomized analysis of variance,and the comparison among groups was performed by the t test.ResuIts The expressions of CD44,CD90,CD106 were positive,and the expressions of CD34,CD45 and CD11b were negative.No insulin DNA transcription was detected in the control group and vacant vector group.The levels of insulin secreted were (4.7 ± 0.8) mIU/L,(8.8 ± 0.5) mIU/L,(8.9 ± 0.8)mIU/L,(8.6 ± 0.6)mIU/L in the recombinant vector 1,6,12,18 days group,which were significantly higher than (1.3 ± 0.6) mIU/L in the control group and (1.7 ± 0.8) mIU/L in the vacant vector group (t =10.09,32.64,22.20,55.53 ; 9.23,27.56,19.43,51.25,P ＜ 0.05).There were significant differences in the levels of insulin secreted between the recombinant vector 1 day group and the recombinant vector 6,12,18 days groups (t =12.77,12.26,13.93,P ＜0.05).There were no significant difference in the levels of insulin secreted between the recombinant vector 6,12,18 days groups (F =45.67,P ＞ 0.05).There was a significant difference in the level of insulin secreted between the high glucose group and the low glucose group (t =2.03,P ＜ 0.05).The result of the glucose stimulation test was negative.Conclusion The ADMSCs are transfected into insulinsecreting cells in vitro successfully,and the secretion of insulin is stable.Although the secretion of insulin can't change in line with the concentration of glucose,it is a new seed cell for the treatment of diabetes with stem cells.

Objective To detect nucleos(t)ide-resistance mutations in hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) isolated from hepatocytes of patients with chronic HBV infection and to analyze the correlation between the mutations found in cccDNA and relaxed circular DNA (rcDNA). MethodsForty patients with chronic HBV infection were investigated. Preoperation serum samples and non-tumor liver tissues were collected.Intrahepatic HBV cccDNA and rcDNA were selectively extracted by co-precipitation of sodium dodecyl sulphate-protein and QIAamp DNA Mini Kit, and further purification with plasmid-safe ATP-dependent DNase (PSAD).Thereafter,cccDNA were amplified by selective polymerase chain reaction (PCR) or nested PCR using the primers spanning both the two gaps in HBV genome and covering the common mutations associated with nucleoside analogues resistance (rt169- rt250).Intrahepatic HBV rcDNA and pre-operation serum HBV rcDNA were also extracted and then amplified by PCR.The PCR products were then purified and sequenced.Results Among the 40 patients,intrahepatic HBV cccDNA were detected in 31 patients,and HBV rcDNA were detected in liver samples of 35 patients and pre-operation serum samples of 21 patients. The PCR products amplified from these samples were all successfully sequenced.rtM204Ⅰ mutation was detected in intracellular HBV cccDNA,rcDNA and serum HBV rcDNA in 2 patients.Both rtM204Ⅰ and rtQ215H were detected in intrahepatic HBV cccDNA and rcDNA in 2 patients,while no corresponding mutation was observed in serum HBV rcDNA of these two patients.Three variants including rtM204V,rtM204V and rtV173L-rtL180M-rtM204V were detected in serum HBV rcDNA in 3 patients,while corresponding mutants were not detected in intracellular HBV cccDNA or rcDNA of these patients.Condsions The results suggest that antiviral nucleos(t) ide resistance mutations can be found in HBV cccDNA in chronic hepatitis B patients. The dominant resistant mutation found in intrahepatic HBV cccDNA/rcDNA may be different from that in serum HBV rcDNA.

Objective To determine whether hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) could be detected in serum of patients with hepatitis B and evaluate the related factors and clinical significances. Methods Fifty-seven patients, including 26 with mild chronic hepatitis B (CHB) and 31 with severe hepatitis B (SHB) were enrolled. Prothrombin time (PT), hepatic biochemical indexes, serum markers of hepatitis virus, serum total HBV DNA and HBV cccDNA of every patient were detected after hospitalization. Factors associated with the detection rate of serum HBV cccDNA were analyzed using Logistic stepwise regression. Results Serum HBV cccDNA was detected in 13 patients with SHB and 1 with mild CHB, and serum levels of HBV cccDNA were varying from 1.25 × 103 to 4. 88 × 104 copy/mL. The detection rates were significantly different between the two groups (P=0. 0014). The sensitivity and specificity of SHB diagnose by serum HBV cccDNA detection were 41.94% and 96.15 %, respectively. Logistic regression analysis showed that the detection rate of serum HBV cccDNA was associated with PT (X2 = 7. 2192, P= 0. 0072), while not associated with age, sex, total serum HBV DNA, total bilirubin or alanine aminotransferase (ALT). Conclusion Serum HBV cccDNA could be detected in some of the patients with SHB, whic hmay be considered as one of the diagnostic indexes for SHB,

Objective To investigate the coordinate repression of angiostatin(AS)and Fas gene on human colon carcinoma LOVO.Methods Plasmid pcDNA3-AS and pcDNA3-Fas were constructed,and AS,Fas,AS and Fas gene were transfected to human colon carcinoma LOVO cells by liposome method.The expressions of target protein were detected by Western blot.The effects of transfection of AS and Fas gene on the growth of human colon carcinoma cells were detected by MTr methods.AS,Fas,AS and Fas gene were transfected to the human colon carcinoma subcutaneously implanted in nude mice by direct injection into the tumor.The tumor sizes were detected after 14 days of the first gene transfection.Results The effect of gene transfeetion in LOVO cell after twelve hours,24 h,48 h and 72 h were observed and compared with control group,co-transfection of Fas and AS gene group and Fas group significantly inhibited the growth of human colon carcinoma LOVO line cells(P＜0.01).Fourteen days after plasmid transfection,the tumor volume in group Fas and group co-transfection were signifieandy smaller than that of control,AS and Fas group(P＜0.05).The tumor volume in AS group and Fas group were significantly smaller than that of control(P＜0.05).Conclusion Angiostatin and Fas gene have coordinate repression effect on human colon carcinoma in vivo.