Objective To establish a new approach for quantitative detection of VAD1 mRNA in cryptococcus neoformans by RT-FQ-PCR, and evaluate the treatment efficacy of CNM. MethodsThe primers and TaqMan probe were designed according to the published sequence of VAD1 mRNA (GenBank),and RT-FQ-PCR method to detect VAD1 mRNA was established. Cerebrospinal fluid from 25 CNM patients and 30 controls were detected and sensitivity and specificity of the method were evaluated. VAD1 mRNA concentration in cerebrospinal fluid from both acute phase and recovery phase of 25 CNM patients were also detected and significance of CNM treatment efficacy with VAD1 mRNA analysis was evaluated. Results Correlation coefficient of standard curve was - 0. 997 9 in detection of VAD1 mRNA by RT-FQ-PCR, and the detection limit was 101 copies/μl. The intra CV of plasmid standard for high, medium and low concentrations were 0. 65% ,0. 89% and 1.23% respectively, the sensitivity of cryptococcus neoformans detection by RT-FQPCR was 96% (24/25) ,while specificity was 100% (30/30). VAD1 mRNA concentration in acute phase were significant higher than that in recovery phase (3. 042 ±0. 906 vs 2. 187 ±0. 665 ,t =4. 583 ,P ＜0. 01).Conclusions The established RT-FQ-PCR method for the detection of VAD1 mRNA is provided with sound sensitivity, specificity and reproducibility, which might be fit for the detection of VAD1 mRNA. The expression level of VAD1 mRNA is relevant with the treatment efficacy of CNM.

AIM:To study the therapeutic effect of traditional Chinese medicine Reduqing on human cytomegalovirus (HCMV). METHODS:The expression of the HCMV late-mRNA in infected cells was measured by quantitative reverse transcription-polymerase chain reaction(RT-PCR) and the development of cytopathic effect (CPE) caused by HCMV was observed before and after treatment with Reduqing. On the basis of its anti-HCMV activity study in vitro, Reduqing was applied to the child-bearing aged women and pregnant women who were HCMV-IgM positive which indicated the active HCMV infection. The serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2) before and after treatment were measured by the ELISA method. RESULTS:The results showed that the Reduqing had a strong inhibitory effect on both HCMV-mRNA and CPE. The level of the TNF-α increased while IL-2 decreased significantly in HCMV-IgM positive serum when compared with HCMV-IgM negative cases. In HCMV-IgM positive patients, after treatment with Reduqing, the serum levels of TNF-α and IL-2 returned to normal, and 73 % of them turned to be HCMV-IgM negative, which was significantly higher than that (14 %) in control group. CONCLUSION:It is suggested that serum TNF-α and IL-2 level are closely related to the activity of HCMV. Reduqing can enhance the immunity of the body and has an obvious inhibition on HCMV in vitro and in vivo, and can thus serve as an effective medicine for active HCMV infection therapy, especially for the pregnant women.

Objective:To evaluate the clinical efficacy and safety of repaglinide and gliclazide in the treatment of elderly type 2 di-abetes mellitus (T2DM). Methods:A total of 225 T2DM patients with the age over 60 were enrolled and randomly divided into repa-glinide group (n=112) and gliclazide group (n=113). The repaglinide group was given repaglinide 0. 5~2 mg, po, tid, while the gliclazide group was given gliclazide modified release tablets 30~120 mg, po, qd,both with the treatment course of 12 weeks. The level of FPG, 2h PG, HbAlC, TC and TG and the side effects were observed and the clinical efficacy was compared between the two groups. Results:The level of FPG, 2h PG, HbAlc and TC in the two groups showed significant decrease after the treatment (P<0. 01 or P<0. 05), and the level of 2h PG was decreased more significantly in the repaglinide group (P<0. 05). There were no significant differ-ences in the total effective rate (91. 8% vs 86. 2%), hypoglycemia incidence (4. 5% vs 8. 2%) and ADR incidence (6. 4% vs 10. 1%) in the two groups (P>0. 05). Conclusion:Both repaglinide and gliclazide show safe and reliable efficacy in the treatment of elderly T2DM.

OBJECTIVETo observe effects of mild moxibustion and lovastatin on immunologic function in rabbits with chronic hyperlipidaemia and atherosclerosis (AS) to initially explain regulating rules of mild moxibustion on immunologic function.

METHODSAmong thirty-two Japanese male big-ear rabbits, 8 rabbits were randomly selec ted as a blank group, the rest 24 rabbits were fed with method of endothelial injury and high-fat diet to establish AS model. The blank group was raised with normal diet and free water. After ten weeks of model establishment, the rest 24 rabbits were randomly divided into a model group, a moxibustion group and a medicine group, eight rabbits in each one. Moxibustion was applied at "Shenque" (CV 8) and "Zusanli" (ST 36) for 10 min per acupoint per day in the moxibustion group, while intragastric administration of 3.6 mg/kg lovastatin capsule was applied in the medicine group. After treatment, serum was acquired. Spectrophotometry method was adapted to measure cholesterol (TC) and high-density lipoprotein (HDL-C) and evaluated atherosclerosis index (AI), while enzyme-linked immunosorbent assay method was used to measure interferon-gamma (IFN-gamma) and interleukin-4 (IL-4).

RESULTS(1) The serum TC and HDL-C in the model group were significantly higher than those in the blank group, moxibustion group and medicine group (all P < 0.01). The mean value of AI was 1.683 +/- 0.486 in the moxibustion group, which was obviously lower than 20.301 +/- 4.022 in the model group (P < 0.01). (2) The ratio of Th1/Th2 was 0.569 +/- 0.143 in the moxibustion group and 0.445 +/- 0.079 in the medicine group, which were significantly lower than 0.917 +/- 0.255 in the model group (both P < 0.01), but there was no statistically significant difference between the moxibustion group and the medicine group (P > 0.05).

CONCLUSIONThe moxibustion for AS could reduce atherosclerosis index, influence drift and bias of helper T cell and regulate balance between humoral immunity and cellular immunity. As a result, status of relative balance of immunity is acquired, which could slow down the development of atherosclerosis and process of thrombus burst.

Animals ; Atherosclerosis ; immunology ; therapy ; Humans ; Hyperlipidemias ; immunology ; therapy ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Male ; Moxibustion ; Rabbits ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVETo explore effects and action mechanism of warm moxibustion on regulation of blood lipids and anti-atherosclerosis.

METHODSForty-one male Japanese big-ear white rabbits were randomly divided into a blank group (10 rabbits), a model group (10 rabbits), a moxibustion group (10 rabbits) and a medication group (11 rabbits). Normal diet was applied in the blank group while high-cholesterol diet combined with injection of bovine serum albumin were applied in the rest groups to establish rabbit model of atherosclerosis. After establishment, the model group was not intervened and warm moxibustion was applied in the moxibustion group at "Zusan-li" (ST 36) and "Shenque" (CV 8), 10 min per acupoint per day for continuous 4 weeks. The medication group was treated with intragastric administration of lovastatin capsule (3.6 mg/kg) for continuous 4 weeks. The level of blood lipids, such as total cholesterol (TC), triglyceride (TG), and content of CD40 ligand (CD40L), soluble CD40 ligand (sCD40L) and expression of nuclear factor NF-kappaB were tested after 4 weeks.

RESULTSCompared with the model group, the moxibustion group and medication group could effectively reduce the contents of TC and low density lipoprotein (all P < 0.05), lower the level of sCD40L [(8.310 +/- 1.221) ng/mL in the model group, (7.097 +/- 0.846) ng/mL in the moxibustion group and (7.354 +/- 0.631) ng/mL in the medication group], reduce expression of CD40L [(0.235 +/- 0.179) mm2 in the model group, (0.072 +/- 0.079) mm2 in the moxibustion group and (0.039 +/- 0.015) mm2 in the medication group] and NF-kappaB [(0.145 +/- 0.052)mm2 in the model group, (0.052 +/- 0.012) mm2 in the moxibustion group and (0.036 +/- 0.013) mm2 in the medication group], indicating the significant difference (P < 0.05, P < 0.001). There was no statistical difference between the moxibustion group and medication group (all P > 0.05).

CONCLUSIONThe warm moxibustion has great effect on regulation of blood lipids and anti-atherosclerosis, in which lowering expression of CD40-CD40L could be one of possible mechanisms to take effect of anti-atherosclerosis.

Animals ; Atherosclerosis ; blood ; therapy ; CD40 Antigens ; blood ; CD40 Ligand ; blood ; Humans ; Lipids ; blood ; Male ; Moxibustion ; methods ; Rabbits

Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid (CSF) were recruited and patients with craniocerebral trauma were recruited as controls.Standard plasmids were constructed using standard CN strain.Real time fluorescent quantitative polymerase chain reaction (RT-FQ-PCR) was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis,which were compared with the results of CSF ink staining,fungal culture and antigen detection.The diagnostic values of single gene test and combined gene test were compared by chi square test.Results Among 23 CN meningitis patients,22 (95.6％) were CAP10 mRNA positive detected by RT-FQ-PCR,which was significantly higher than both ink staining (16/23,69.6％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).There were also no statistical significant differences between combined detection of CAP 10 + VAD1 and CAP 10 or VAD1 single gene test (P＞0.05).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.

OBJECTIVE:To study the in vivo antibacterial effects of Norfloxacin injection in animals.METHODS:After infecting the animals by injection of Escherichia coli and Staphylococcus aureus suspension into the mice's abdominal cavity and rabbit's body,the animals were divided into therapy,prophylaxis and control groups to observe the effect of Norfloxacin.RESU-LTS:All mice of control group died within 24 hours after the injection of bacteria while those of the therapy and prophylaxis groups survived.The results of experiment on rabbits were similar to those on mice.CONCLUSION:The results show that Norfloxacin injection has obvious therapeutic and prophylactic effects on infections induced by E.coli and Staphylococcus aureus.

Objective To discuss the effect of “practice teaching methods ” in musculoskeletal ultrasound diagnosis training process .Methods Thirty refresher doctors for a month with the method training .The effect training is evaluated by objective exam and questionnaire survey .Results Thirty refresher doctors received tests before and after the training .Test results showed that the accuracy after the training was significantly higher than that of before ( P<0.05 ) .Refresher doctors survey about satisfaction of the training method showed that whole satisfaction is 100%, in which 70%were very satisfied , 30%were satisfied .The self-assessment results for the improvement of the level of diagnosis musculoskeletal ultrasound showed that 45%doctors are very satisfied ,40%doctors are satis-fied, and 15%is not satisfied .Conclusions With the implementation of “teaching by practice” method, refresher doctors could master the knowledge of musculoskeletal anatomy as soon as possible , and have the ability to diagnose common musculoskeletal diseases independently .

Amygdala ; drug effects ; metabolism ; Animals ; Conditioning (Psychology) ; Morphine ; adverse effects ; Rats ; Receptors, Opioid, kappa ; metabolism

Objective: To observe the effects of acupoint thread-embedding therapy and low-carbohydrate diet therapy on obese patients with food addiction. Methods: Sixty-five eligible patients were randomized into a thread-embedding group of 33 cases and a diet group of 32 cases to respectively receive 12-week treatment. Before treatment, after treatment and at 6-month follow-up, the two groups were observed and compared in terms of body mass (BM), waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), body mass index (BMI), body fat rate (BFR), basal metabolic rate (BMR) and Yale food addiction scale version 2.0 (YFAS 2.0). Results: At the end of treatment, there were no significant differences in the general efficacy, and the improvements in BM, BMI, WC, HC, WHR and BFR between the thread-embedding group and diet group (all P>0.05). At follow-up, the thread-embedding group showed more significant improvements in all the aforementioned indicators compared with the diet group except HC (all P<0.05). At the end of treatment and follow-up, BMR and YFSA 2.0 had more significant improvements in the thread-embedding group than in the diet group (all P<0.05). Conclusion: Acupoint thread-embedding therapy can produce significant efficacy in treating obese patients with food addiction; it can improve the food addiction state and work better in maintaining the efficacy compared with low-carbohydrate diet therapy.