The ribosomal DNA fragment was amplified with PCR utilizing synthetic primers with host bacteria chromosomal DNA as template. The resultant probes were labeled with Digoxigenin and were applied in the dot blot of DNA vaccine samples. The host bacteria genomic DNA in DNA vaccine against hepatitis B was less than 15 ng/?g. Digoxigenin labeled probes proved sensitive and reliable in determining genomic DNA.

Objective To evaluate the inflammatory response and the expressions of peroxisome proliferator-activated receptor(PPAR)isoforms(PPARα,PPARβ/δ,and PPARγ)in the brain(cortex,striatum,cerebellum)of spontaneously hypertensive rats(SHR).Methods Brain tissues(cortex,striatum,and cerebellum)were dissected from SHR and age-matched control Wistar-Kyoto rats.Myeloperoxidase(MPO)activity was measured in brain tissues as an index of neutrophil accumulation and the carbonyl protein content was analyzed by spectrophotometry to evaluate the protein oxidation.RT-PCR and Western blotting were performed to examine the expressions of inflammatory mediators(IL-1β,TNFα,ICAM-1,and iNOS)and nuclear factor PPARs(PPARα,PPARβ/δ,and PPARγ),respectively.Results(1)Systolic blood pressure of SHR was significantly higher than that of Wistar-Kyoto rats,(205.4±9.4)mm Hg versus(130.4±7.9)mm Hg(t=14.96,P＜0.01).(2)MPO activity of cortex,striatum,and cerebellum were markedly higher in SHR than in Wistar-Kyoto rats.Carbonyl protein levels of cortex,striatum,and cerebellum in Wistar-Kyoto rats and SHR were(3.27±0.43)nmol/mg versus(11.87±1.11)nmol/mg,(4.02±1.04)nmol/mg versus(14.06±1.36)nmol/mg,(5.94±0.71)nmol/mg versus(14.95±1.82)nmol/mg,indicating significantly higher levels of protein oxidation in SHR than Wistar-Kyoto rats(t=17.70,14.36,11.30,P＜0.05).Consistently,the expression of pro-inflammatory mediators(IL-1β,TNFα,ICAM-1,and iNOS)was upregulated when compared with Wistar-Kyoto rats.The difference between SHR and control Wistar-Kyoto rats was statistically significant except the mRNA expression of IL-1β in striatum,cerebellum and TNFα in cerebellum of SHR.All the above experimental data indicated the occurrence of inflammatory status in the brain tissue of hypertension.(3)mRNA and protein levels of brain PPAR isoforms(PPARα,PPARβ/δ,and PPARγ)of SHR increased significantly when compared with Wistar-Kyoto rats.Specifically.protein levels of PPARα in cortex.striatum,and cerebellum of SHR increased by 644.78%,791.95%,and 42.85%;PPARβ/δ increased by 106.72%,94.12%,and 161.44%;PPARγ was up-regulated by 2700.16%,790.81%,and 875.00%compared with that of Wistar-Kyoto rats,respectively.Conclusions The brain(cortex,striatum,and cerebellum)from SHR shows marked inflammatory status and increased expression of all PPAR isoforms.Increases in PPARs expression may play a compensatory role in the inflammatory response of the brain in SHR.

OBJECTIVE: To study clinical characteristics, diagnosis, pathological types and therapy methods of nonsquamous cell neoplasm in subglottis, in order to improve comprehension of this rare disease. METHOD: To analysis 3 patients' clinical data with noosquamous cell neoplasm in subglottis by reviewing related literature, and to generalize the symptoms, differentiation and treatment experience of nonsquamous cell neoplasm in subglottis. RESULT: Among the 3 patients, one's main complain was hoarseness and the other two's main symptom was inspiratory dyspnea. As diagnosis was definite and acute obstruction of airway was relieved, the neoplasms were removed totally. Two patients were undergone operation through oral cavity by using suspended laryngoscope, and one patient was done surgery by neck approach. Paraffin wax result showed that one patient suffered epithelial-myoepithelial carcinoma, who received radiotherapy after surgery. There is no recurrence in all patients by following up more than 1 year. CONCLUSION The occurrence of nonsquamous cell neoplasm in subglottis is relatively low, and the subglottis should be inspected carefully to avoid missed diagnosis. Operation is the preferred method to treat this kind of disease. To choose a suitable operational manner according to size, location of the neoplasm is crucial.
Female ; Glottis ; pathology ; Humans ; Laryngeal Neoplasms ; diagnosis ; pathology ; surgery ; Male ; Middle Aged

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Female ; Hemifacial Spasm ; therapy ; Humans ; Male ; Meridians ; Middle Aged

Objective To investigate effect of artesunate on expressions of Toll-like receotor-4 (TLR4) and interleu?kin-8 (IL-8) in renal tissue of diabetic nephropathy rats. Methods Male SD rats of over 200 g in body weight (n=45) were injected with low dosage streptozotocin and fed with high fat and sucrose diets to establish diabetes nephropathy (DN) model. Among all rats of successful model, thirty-two were randomly selected and divided into four groups (n=8 in each group), Mod?el group (Group B), Artesunate at high dose treatment group [30 mg/(kg · d)] (Group C), Artesunate at low dose treatment group [10 mg/(kg·d)] (Group D), Enalapril treatment group [10 mg/(kg·d)] (Group E). Another eight rats without STZ injec?tion whose body weight is under 200 g were assigned as Normal group (Group A). Then, rats in Group A and B were given the same volume of normal saline [10 mg/(kg·d)] while rats in Group C, D and E were given 30 mg/(kg·d) Artesunate, 10 mg/(kg·d) Artesunate and 10 mg/(kg · d) Enalapril respectively intragastrically for consecutive 12 weeks. Fasting blood glucose, body weight, kidney index, 24-hours proteinuria, serum creatinine (Cr) and blood urea nitrogen (BUN) were measured. Expression level of TLR4 in renal tissue of rats were examined by Western blot;ELISA was used to quantify the concentration of IL-8 in serum and renal tissue of rats. Results Compared with rats in Group A, the levels of Fasting blood glucose, kidney index , 24-hours proteinuria, Cr and BUN as well as the level of TLR4 in kidney, levels of IL-8 in serum and kidney all significant?ly increased in rats in Group B, C, D and E(P<0.05). On the other hand, body weight were decreased in rats of Group B, C, D and E compared to rats in Group A(P<0.05). The level of TLR4 in kidney, levels of IL-8 in serum and kidney, 24-hours proteinuria, Cr and BUN in rats of Group C, D and E were significantly lower than those in rats of Group B(P<0.05). Furthermore, compared with rats in Group D, the levels of TLR4 in kidney, IL-8 in serum and kidney, 24-hours proteinuria, Cr and BUN were decreased in rats in Group C and E(P<0.05). Pearson correlation analysis showed that the expression lev?el of TLR4 positively correlated with IL-8 level in serum (r=0.969, P<0.05), IL-8 level in kidney (r=0.972, P<0.05), 24-hours proteinuria(r=0.965, P<0.05), Cr(r=0.944, P<0.05)and BUN(r=0.903, P<0.05). IL-8 level in kidney positively correlated with 24-hours proteinuria(r=0.962, P<0.05), Cr(r=0.929, P<0.05)and BUN(r=0.940, P<0.05). Conclu?sion Artesunate decreases expression of TLR4 and IL-8, reduce 24-hours proteinuria, inhibits the inflammation of the DN,relives the pathological lesions of nephron rats with diabetic nephropathy.

By summarizing the recent literatures on brain mechanisms with acupuncture intervention based on blood oxygenation level-dependent (BOLD)-functional magnetic resonance imaging (fMRI), the BOLD-fMRI examination and analysis methods, the points to be acupunctured, the corresponding meridian activation regions, the specific intensity range, functions and indications of the acupoints, the manifestation of 'bi-directional regulation' characteristics, fMRI performance of chronergy, laterality and needling qi of acupuncture were reviewed to provide the ideas for future research in this area.

To study preliminarily the effect of Jiawei Bazhen decoction combined with oxytocin in promoting cervical ripening of full-term pregnancy women who were in the deficiency of qi and blood type through the syndrome differentiation of traditional Chinese medicine (TCM). 180 patients that met the inclusion criteria of the study were randomly divided into three groups: the control group(oxytocin group), the treatment group (Jiawei Bazhen decoction combined with oxytocin group), the blank control group (expected and observation group). Cervical maturity score (Bishop score), vaginal and cervical secretions fetal fibronectin (FFN), the result of induced labor, the result of mother and baby were observed in each group before and after treatment. The result comes out that the cervical Bishop score of pregnant women for treatment group were significantly higher than the control group and blank control group after treatment (P < 0.05). The FFN of pregnant women for the treatment group were significantly different from the control group and blank control group after treatment (P < 0.05). The pregnancy outcome of the three groups: the labor rate and rate of vaginal delivery of the treatment group were higher than the other two groups, and the difference was statistically significant (P < 0.05). The cesarean section rate of the treatment group was significantly lower than the other two groups, the difference was also statistically significant (P < 0.05). The three groups did not appear the phenomenon of neonatal asphyxia. Jiawei Bazhen decoction combined with oxytocin is effective in producing cervical ripening and induce labor. It is convenient, safe and reliable, for it is no obvious adverse effects on mother and fetus, but effective in reducing the rate of cesarean section, and playing a positive role in promoting natural delivery.
Adult ; Cervical Ripening ; drug effects ; metabolism ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fibronectins ; secretion ; Humans ; Labor, Induced ; Oxytocin ; administration & dosage ; Pregnancy ; Pregnancy Complications ; drug therapy ; metabolism ; physiopathology ; Pregnancy Outcome ; Qi ; Vagina ; drug effects ; secretion ; Young Adult

Anti-Bacterial Agents ; therapeutic use ; Antigens, CD20 ; metabolism ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Fever ; drug therapy ; virology ; Herpesvirus 6, Human ; isolation & purification ; Humans ; Lymph Nodes ; pathology ; virology ; Lymphadenitis ; drug therapy ; metabolism ; pathology ; virology ; Male ; Middle Aged ; Roseolovirus Infections ; drug therapy ; metabolism ; virology

Objective To investigate the effects of high-fat diet induced insulin resistance on fibroblast growth factor-21 (FGF-21) and its receptors expression in ApoE~(-/-) mice. Method Male ApoE~(-/-) mice were randomly divided into normal-chow group(NF,n=20)and high-fat fed group(HF,n=20) and fed for 16 weeks. The insulin sensitivity and glucose-lipid metabolism in awake mice were evaluated by hyperinsulinemic-euglycemic clamp technique combined with 3-[~3H]-glucose as a tracer. The Mrna expressions of FGF-21,β-klotho, and FGFR1-4 were measured by quantitative real-time PCR. FGF-21 protein levels were determined by Western blot. Results Fasting blood glucose, plasma insulin and free fatty acids, triglycerides, free fatty acids, and cholesterols were significantly elevated in HF group compared with NF group(all P<0.01). During the steady-state of clamp, plasma insulin was significantly higher in HF group than that in NF group(P<0.01), and glucose infusion rate was also significantly decreased(P<0.01). At the end of insulin clamp, glucose disappearance rate was significantly lower in HF group than that in NF groups(P<0.01). Hepatic glucose production in NF group was suppressed by 70% ,while in HF group it was suppressed by 51%. The FGF-21 Mrna expressions of hepatic and adipose tissues in HF group were significantly increased compared with NF group(both P<0.01), and β-klotho Mrna expressions increased(P<0. 05). In hepatic and adipose tissues, FGFRI, Mrna expressions were higher in HF group than those in NF group(both P<0.01) ,and FGFR3 Mrna increased(P<0.01 and P<0.05, respectively). In hepatic tissue,FGFR4 Mrna levels were significantly up-regulated in HF group(P<0. 05). Plasma FGF-21 levels were elevated in HF group compared with NF group(P<0.01) ,and FGF-21 protein expressions of hepatic and adipose tissues were also increased(both P<0.05). Conclusion FGF-21, β-klotho, FGFR1, and FGFR3 were significantly up-regulated in ApoE~(-/-) mice fed by high-fat diet, and they might be the targets in regulating glucose-lipid metabolism by FGF-21.

Objective To study the protective mechanism of dl-3-n butylphthalide (NBP)on oxidative stress injury induced by hydrogen peroxide(H2O2)in rat bone marrow stem cells(rBMSCs).Methods The rBMSCs were divided into control,H2O2 and different concentration NBP pretreatment groups.The control group received no treatment.An oxidative stress injury model was induced by H2O2 for 4 hours with the final concentration 600 μmol/L in the H2O2 group.In the NBP pretreatment groups,the rBMSCs were pretreated with different NBP concentrations(0.1,1,10,and 100 μmol/L)for 24 hours,then treated with H2O2 for 4 hours with the final concentration 600 μmol/L.The cell viability was detected by MTT method.The apoptosis rate was detected by flow cytometry.Superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were measured with SOD and MDA commercial kits.Results The cell activity(A)was 0.487 ±0.018 in the H2O2 group,and it was significantly lower than 0.750 ±0.016 in the control group(P =0.000);they were 0.597 ±0.024,0.666 ±0.033,and 0.658 ±0.012 in the NBP 1,10,and 100 μmol/L pretreatment groups,they were all significantly higher than the H2O2 group(all P =0.000),and showed a dose dependent manner.The apoptosis rate was(44.96 ± 2.84)% in the H2O2 group,and it was significantly higher than (0.15 ±0.07)% in the control group(P= 0.000).The apoptosis rates were(31.79±1.60)% 、(21.41 ± 1.92)% and(22.59 ± 1.78)% in the NBP 1,10,and 100 μmol/L pretreatment groups,they were all significantly lower than the H2O2 groups(all P= 0.000),and showed a dose-dependent manner.The SOD activity was(24.01 ± 2.85)U/mg in the H2O2 group(P = 0.000),and it was significantly lower than(43.58 ± 2.72)U/mg in the control group(P =0.000);they were(28.29 ± 1.19),(34.06 ± 1.83),and(31.76 ± 1.75)U/mg in the NBP 1,10,and 100 μmol/L pretreatment groups,and they were all significantly higher than the H2O2 group(all P = 0.000).The MDA content was(7.98 ± 0.55)nmol/mg in the H2O2 group,and it was significantly higher than(4.73 ± 0.53)nmol/mg in the control group(P =0.000);they were(6.97 ±0.29),6.09 ±0.28),and(6.15 ±0.41)nmol/mg,respectively in the NBP 1,10,and 100 μmol/L pretreatment groups(P = 0.000),they were significantly lower than the H2O2 group,and showed a dose-dependent manner.Conclusions NBP has obvious protective effects on oxidative stress injury induced by H2O2 in rBMSCs.Its mechanism may be associated with the role of antioxidant oxidative stress of NBP.