OBJECTIVES: This study aimed to demonstrate the activities and phosphorylation changes induced by acute ethanol treatment and withdrawal conditions in the intracellular signal transduction molecules [such as extracellular signal-regulated kinase (ERK), glycogen synthase kinase 3beta (GSK3beta), and Akt] of the SH-SY5Y neuroblastoma cell line. METHODS: The acute treatment exposed SH-SY5Y cells to 100 mM ethanol, and we took samples 30 minutes, 60 minutes, and 24 hours after initiating this treatment. After 24 hours' continuous ethanol treatment, we initiated ethanol withdrawal, taking samples at 30 minutes and 60 minutes. We assayed the kinase phosphorylations via an immunoblot analysis using phosphorspecific antibodies, quantified by optical densitometry. RESULTS: Ethanol treatment induced a transient increase in phosphorylation of GSK3beta and Akt at 30 minutes but failed to change the phosphorylation level of ERK. Ethanol withdrawal induced a transient ERK phosphorylation increase at 30 minutes, but it had no effect on the phosphorylation of GSK3beta or Akt. CONCLUSION: The results indicate that the ethanol-induced cellular response includes the ERK, GSK3beta, and Akt systems. In particular, the ERK pathway may play a role in the acute withdrawal response. This also suggests that a relatively short exposure to ethanol, such as the 24-hour exposure in this study, can induce functional adaptation within a cell.
Antibodies ; Cell Line ; Densitometry ; Ethanol ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinases ; Humans ; MAP Kinase Signaling System ; Neuroblastoma ; Phosphorylation ; Phosphotransferases ; Signal Transduction

OBJECTIVETo compare differences of clinical therapeutic effect of heat-sensitive moxibustion with different doses for irritable bowel syndrome (IBS).

METHODSSixty cases of IBS were randomly divided into a saturated-dose group (30 cases) and a traditional-dose group (30 cases). Heat-sensitive moxibustion was applied in both groups. The acupoints that had the strongest heat-sensitive feeling were selected and treated by warm and suspended moxibustion with moxa stick. Disappearance of heat-sensitive feeling was taken as the sign of treatment time in the saturated-dose group, while the traditional-dose group was treated for 15 min each time. The treatment in both groups was given twice a day for first 5 days, and from the sixth day it was given once a day for continuous 25 times, totally 30 days. Clinical symptom scores and therapeutic effect before and after treatment in two groups were observed.

RESULTSAfter the treatment, the cured and markedly effective rate was 75.0% (21/28) in the saturated-dose group, which was inferior to 44.4% (12/27) in the traditional-dose group (P < 0.05). The clinical symptom scores, including diarrhea, abdominal distension and pain, were obviously reduced in two groups compared with those before the treatment (all P < 0.05). Compared with the traditional-dose group, the symptom scores of diarrhea and abdominal distension in the saturated-dose group were obviously decreased (0.87 +/- 0.13 vs 1.27 +/- 0.21, P < 0.01; 1.12 +/- 0.41 vs 1.32 +/- 0.26, P < 0.05).

CONCLUSIONThe moxibustion featured with disappearance of heat-sensitive feeling and saturated dose has better therapeutic effect than that with traditional-dose for treatment of IBS. As individual dynamic amount of moxibustion, disappearance of heat-sensitive and quantitation varies from person to person, which is one of the key factors to obtain the best curative effect.

Acupuncture Points ; Adult ; Female ; Humans ; Irritable Bowel Syndrome ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

Objective To study the changes of plasma cystatin C level (PcyC),and evaluate the effects of the joint use of atorvastatin and probucol on PcyC and severity of coronary lesion in patients with borderline lesion of coronary artery.Methods One hundred and thirty consecutive patients with borderline coronary lesion assessed by quantitative coronary angiography were enrolled into borderline coronary lesion group (BCL),and another 136 subjects without coronary lesion were enrolled as controls (CTR).And in the meantime,the subjects in BCL group were randomized (closed envelope method) into routine treatment subgroup ( RTT,n =60),and combined treatment subgroup in which patients were treated with atorvastatin 20 mg plus probucol 1.0 g daily in addition to routine medication ( CBT,n =70) for 6 months.There were no statistical differences in basic clinical features between two subgroups.PcyC,high-sensitive C-reactive protein (hs-CRP),total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol ( HDL-C ) and triglycerides (TG) were determined.Of them,104 patients in BCL group rechecked by coronary angiography.Comparison of biomarkers carried out between two groups by using a number of independent-sample t-test and analysis of variance.For enumeration data,chi-square test was used to compare mean values of biomarkers between groups. P ＜ 0.05 was considered statistically significant.Results PcyC levels were significantly higher in BCL group than those in CTR group ( P ＜ 0.05 ).Compared with RTT subgroup,levels of PcyC,TC,LDL-C,TG and hs-CRP were more significantly decreased in CBT subgroup (P ＜ 0.05,P ＜ 0.01 ).Moreover,there was a trend of slight decrease in the mean percent of stenosis (MPS) of coronary artery with borderline lesion in RTT subgroup treated for 6 months,whereas more marked decrease in the MPS of coronary artery with borderline coronary lesion in CBT subgroup treated for 6 months ( P ＞ 0.05 ; P ＜ 0.05 ).Conclusions Cystatin C plays an important role in the pathogenesis of coronary artery,and PcyC is associated with severity of coronary lesion,the combination of atorvastatin and probucol decreases the PcyC level,and it may be the treatment of choice for borderline lesion of coronary artery.

Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.

Objective To explore the correlation between serum hepatitis B virus (HBV) X antigen/antibody (HBxAg-wild/HBxAb-wild,and HBxAg-mutant/HBxAb-mutant) and the disease progression in patients with chronic HBV infection.Methods A direct enzyme immunosorbent asssay (ELISA) was performed to detect HBxAb using recombinant antigen,and a double antibody sandwich ELISA assay to detect HBxAg using monoclonal antibody and specific rabbit polyclonal antibody.HBxAg-wild/HBxAb-wild and HBxAg-mutant/HBxAb-mutant were tested in sera from cases at different stages of chronic HBV infection.A chi-square test was employed to examine statistical significance.Results The positive rates of HBxAg-wild and HBxAg-mutant in the chronic asymptomatic HBV carriers,chronic hepatitis,hepatitis B-related cirrhosis and liver cancer were 6.2％ (2/32),10.7％ (3/28),28.6％ (6/21),43.6％ (17/39) and 3.1％ (1/32),10.7％ (3/28),33.3％ (7/21),48.7％ (19/39),respectively.The positive rates of HBxAb-wild and HBxAb-mutant in the above mentioned groups were 6.2％ (2/32),21.4％ (6/28),38.1％ (8/21),53.8％ (21/39)and 6.2％ (2/32),25.0％ (7/28),42.9％ (9/21),61.5％ (24/39) respectively.The positive rates of HBxAg-wild and HBxAg-mutant were not significantly different among the above groups (x2 =0.871,0.780,0.565 and 0.317,respectively; all P＞0.05) ; The positive rates of HBxAb-wild and HBxAb-mutant were also similar among all the groups (x2 =0.780,0.709,0.580 and 0.210,respectively; all P＞0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in patients with low viral loads (HBV DNA＜1 × 104 copy/mL) were 36.5％ (23/63),44.4％ (28/63),42.9％ (27/63) and 54.0％ (34/63),respectively,those in patients with high viral loads (HBVDNA≥1×104 copy/mL) were 8.8％ (5/57),15.8％ (9/57),5.3％ (3/57) and 14.0％ (8/57),respectively.The positive rates of HBxAg and HBxAb were significantly higher in cases with low viral loads than those with high viral loads (x2 =12.869,11.522,22.556 and 20.976,respectively; all P＜0.05).The positive rates of HBxAg-wild,HBxAb-wild,HBxAg-mutant,HBxAb-mutant in the HBeAg positive group were 21.7％ (18/83),30.1％ (25/83),22.9％ (19/83) and 32.5％ (27/83),respectively,while those in the HBeAg negative group were 27.0％ (10/37),32.4％ (12/37),29.7％ (11/37) and 40.5％ (15/37),respectively.No significant difference of HBxAg/HBxAb positive rates between HBeAg positive group and HBeAg negative group was noticed (x2 =0.408,0.064,0.638 and 0.722,respectively; all P＞0.05).Conclusions The antigenicity and specificity of HBV X protein remains similar after the occurrence of A1762T/G1764A double mutant in X gene.It is also found that the positive rates of HBxAg and HBxAb increase with disease progression.HBxAg/HBxAb might be promoting factors for tumorigenesis in chronic HBV infection.HBxAg and HBxAb might have negative influence on HBV replication.

Objective To observe the diagnostic value of non-invasive 128-slice computed tomography coronary angiography(CTA)in comparison with invasive coronary angiography.Methods 128-slice CTA and invasive coronary angiography were performed in 78 unselected consecutive patients(63 patients with suspected coronary artery disease and 15 patients with previous coronary stenting,56 males,mean age 61±10 years)and ＞50% reduction of minimal lumen diameter was defined as significant coronary stenosis.Results Fifty-eight out of 879 segments(7%)from CTA were not assessable because of irreguldr rhythm,vessel calcification or tachycardia.Compared with invasive coronary angiography,segmentbased analysis from the 821 segments showed the sensitivity by CTA was 87%,specificity 97%,PPV 83% and NPV 97%.Four out of 22 stents implanted in 15 patients were not assessable by CTA because of poor image quality.Compared with invasive coronary angiography,the sensitivity of diagnosing in-stent restenosis by CTA was 100%,specificity 77%,PPV 63% and NPV 100% for the remaining 18 stents-Conclusions One hundred and twenty-eight-slice CTA has a high accuracy for detecting coronary artery disease and instent restenosis after coronary stenting and could be considered as a valuable noninvasive technique for screening coronary artery disease in suspected patients.

OBJECTIVE: Solitaire AB stent-assisted coiling facilitates the endovascular treatment of wide-necked intracranial aneurysms. We present our experience of coiling the micro-aneurysms of wide-neck with Solitaire AB stent assisting in a single center. METHODS: Thirty-one Solitaire AB stents were used to treat via endovascular approach patients with 31 wide-neck micro aneurysms in a single center in China. Technical and clinical complications were recorded. Modified Rankin Scale was used to evaluate the patients' conditions via clinic and telephone follow-up. RESULTS: The mean width of aneurysm sac was 2.30±0.42 mm, and the mean diameter of aneurysm neck was 2.83±0.48 mm. Complete occlusion was achieved in 28 aneurysms (90.32%); neck remnant was seen in 3 aneurysms (9.68%). Technical and clinical complications related to the procedure were encountered in four patients (12.5%). Two patients died (6.25%). No patient had a permanent deficit. CONCLUSION: Solitaire AB stent was a safe and efficiency tool in assisting coiling of micro aneurysms with wide neck, but may be not suitable for a blaster-like one. Mid- and long-term follow-up will be required to elucidate the impact of the Solitaire AB stent on recanalization rate.
Aneurysm* ; China ; Follow-Up Studies ; Humans ; Intracranial Aneurysm ; Neck ; Stents ; Telephone

OBJECTIVETo evaluate the RT-PCR-ELISA method applied for testing live attenuated hepatitis A vaccine titer.

METHODSA solid phase hybridization-enzyme colorimetric detection method was used for detecting specific nucleic acid. Primer labeled with biotin was used to amplify viral gene fragment, then the product was quickly hybridized with the specific probe covalently coupled on DNA-binding microplate wells. Finally, peroxidase-labeled streptavidin was used in colorimetric detection. The results were judged by reading A value. Eleven batches of live attenuated hepatitis A vaccine titer were tested by this method. The results were compared with that of routine cell culture method (CCID50).

RESULTSThe sensitivity was similar to routine cell culture method (P>0.05). This method was convenient, fast and specific.

CONCLUSIONCCID50 method may be replaced by the RT-PCR-ELISA method in evaluating the titer of live attenuated hepatitis A vaccine.

Base Sequence ; DNA, Viral ; analysis ; Enzyme-Linked Immunosorbent Assay ; Genes, Viral ; Hepatitis A Vaccines ; Hepatitis A virus ; genetics ; Quality Control ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Vaccines, Attenuated

OBJECTIVETo observe the therapeutic effect of painless festering moxibustion on knee osteoarthritis and to probe into a new therapy.

METHODSThirty-four cases of knee osteoarthritis were treated with painless festering moxibustion at the best moxibustion part close to the knee joint. The internationally acknowledged knee joint function rating scale (percentage method) including such indexes as pain, walking, joint flexion mobility, instability, engorgement, stair activity and twist-locking before and after treatment was used for assessment of the therapeutic effect.

RESULTSTen cases were cured, 14 cases were markedly effective, 9 cases were effective and 1 cases was ineffective, with a total effective rate of 97.060%.

CONCLUSIONPainless festering moxibustion has an obvious therapeutic effect on knee osteoarthritis.

Adult ; Aged ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Moxibustion ; methods ; Osteoarthritis, Knee ; therapy

OBJECTIVETo study the effect of peroxisome proliferator activated receptor γ (PPAR-γ) agonist on the angiotensin converting enzyme 2 (ACE2) mRNA expression in monocyte-derived macrophages of essential hypertensive patients.

METHODSTotally 57 essential hypertensive patients were randomly divided into three groups: conventional treatment group (n=18), telmisartan group (n=19), and benazepril group (n=20); 20 patients with normal blood pressure were also selected as the control group. Monocyte-derived macrophages were isolated from blood samples of patients in all four groups. The expression of ACE2 mRNA in monocyte-derived macrophages was detected by RT-PCR before treatment and 4 and 12 weeks after treatment.

RESULTSFour and 12 weeks after treatment, the systolic pressure and diastolic pressure of telmisartan group and benazepril group were significantly lower than that of the conventional treatment group (all P<0.01), and the systolic pressure and diastolic pressure of telmisartan group were significantly lower than that of the benazepril group(both P<0.01) .The expression of ACE2 mRNA in monocyte-derived macrophages were significantly lower in essential hypertensive patients than that in control group (P<0.01). After having been treated for 4 weeks and 12 weeks, the expression of ACE2 mRNA in monocyte-derived macrophages of hypertensive patients in telmisartan and benazepril groups were significantly higher than that in conventional treatment group (all P<0.01), and the expression of ACE2 mRNA in telmisartan group was significantly higher than that in benazepril group (both P<0.01).

CONCLUSIONPPAR-γ agonist could increase the ACE2 mRNA expression in monocyte-derived macrophages of essential hypertensive patients.

Aged ; Benzazepines ; pharmacology ; Benzimidazoles ; pharmacology ; Benzoates ; pharmacology ; Female ; Humans ; Hypertension ; drug therapy ; enzymology ; Macrophages ; enzymology ; Male ; Middle Aged ; PPAR gamma ; agonists ; Peptidyl-Dipeptidase A ; genetics ; metabolism ; RNA, Messenger ; genetics