Establishing a Quality Control System of Hedyotis corymbosa

Objective Hedyotis corymbosa(Shui Xian Cao,水线草,HC)is the main antitumor medicine in China and other southeast Asian countries.A comprehensive quality control system of HC was established in this study,as the current system was ineffective.Methods Microscopy was used to observe the characteristics of the powder and cross-sectional tissue.Thin layer chromatography(TLC)was employed to separate HC from Hedyotis diffusa(Bai Hua She She Cao,白花蛇舌草,HD).Simultaneously,high-performance liquid chromatography(HPLC)was used to establish the characteristic spectrum of HC and determine the concentrations of hedyotiscone A.Regarding the issue of systematicness,concentrations of water,total ash,acid-insoluble ash,heavy metals(including lead,arsenic,mercury,cadmium and copper)and pesticide residue were measured by the methods described in the 2015 edition of Chinese Pharmacopoeia.Results Calcium oxalate needle crystal bundles and oil drops were observed in the cross-sectional tissue.Three characteristic blue fluorescent spots were observed in the HC samples by TLC.There were five distinct characteristic peaks in HC but not in HD,and hedyotiscone A was the sharpest characteristic peak.The generated linear equation of hedyotiscone A was Y=3.15×108X+312.04(r2=1.000)between 0.00005270 and 0.002108 mg/mL,and the average recovery was 100.42%.Precision[relative standard deviation(RSD)0.92%],repeatability(RSD 0.14%),and recovery(RSD 0.85%)met the requirements for quantitative analysis.Hedyotiscone A concentrations ranged from 0.00017%~0.00113%across different habitats.Concentrations of water,ash and acid-insoluble ash were 8.80%~11.2%,7.00%~15.3%and 0.440%~7.70%,respectively.Concentrations of Pb,As,Hg,Cd and Cu were 1.60~4.60,0.650~1.60,0.0300~0.100,0.240~0.690 and 10.1~16.6μg/g,respectively.Pesticide residues were less than 0.00100 ppm.Conclusion The quality control system can effectively distinguish HC from HD,so as to comprehensively control the quality of HC,to provide references for the clinical use of HC.

Association of polymorphism of tumor necrosis factor-alpha gene promoter region with outcome of hepatitis B virus infection

AIM： To determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha （TNF-α）, gene promoter and hepatitis B （HB） viral genotypes were associated with outcomes of HBV infection. METHODS： A total of 244 HBV self-limited infected subjects, 208 asymptomatic carriers, and 443 chronic HB patients were recruited to conduct a case-control study. TNF-α -238G/A and -857C/T gene promoter polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）, and HBV genotypes were examined by nested PCR. RESULTS： The positive rate of HBV DNA in asymptomatic carder group and chronic HB group was 46.6% and 49.9%, respectively. HBV genotype proportion among the asymptomatic carriers was 2.1% for genotype A, 25.8% for genotype B, 68.0% for genotype C, and 4.1% for genotype B＋C mixed infection, and 0.9% for genotype A, 21.7% for genotype B, 71.5% for genotype C, 5.9% for genotype B＋C mixed infection in chronic HB group. There was no significant difference in genotype distribution between the asymptomatic carrier group and chronic HB group （X^2 = 1.66, P = 0.647）. The frequency of -238GG genotype in self-limited group was 95.1%, significantly higher than 90.7% in chronic HB group and 89.0% in asymptomatic carrier group （P = 0.041 and P = 0.016, respectively）.The frequency of TNF-α-857 CC in chronic HB group was 79.7%, significantly higher than 64.4% in asymptomatic carrier group and 70.9% in self-limited group （P〈0.001 and P = 0.023, respectively）. A multiple logistic regression analysis revealed that TNF-α-238GA and -857CC were independently associated with chronic HB after gender and age were adjusted.CONCLUSION： TNF-α promoter variants are likely to play a substantial role in the outcome of HBV infection.

Association of-238G/A and -857C/T Polymorphisms of Tumor Necrosis Factor-Alpha Gene Promoter Region With Outcomes of Hepatitis B Virus Infection

Objectives To determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha （TNF-o0 gene promoter were associated with outcomes of hepatitis B virus infection. Methods A total of 246 HBV self-limited infected subjects and 443 chronic hepatitis B （HB） patients were recruited in this case-control study. TNF-α-238G/A and -857C/T gene promoter polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Results The frequency of TNF-α-238 GG （90.7%） in chronic HB group was significantly lower than that （95.1%） in self-limited group （P=0.041）. The frequency of TNF-oc-857 CC （79.7%） in chronic HB patients was significantly higher than that （70.9%） in self-limited infected subjects （P=0.021）. Multiple logistic regression analysis revealed that both TNF-oc-238GA and -857CC were independently associated with chronic HB. Conclusions TNF-α promoter variants are likely to play a substantial role in influencing the outcomes of HBV infection.

Weak type(1, 1) of the Riesz transform on some direct product manifolds with exponential volume growth

In this paper, we are concerned with the Riesz transform on the direct product manifold H^(n)× M,where H^(n) is the n-dimensional real hyperbolic space, and M is a connected complete non-compact Riemannian manifold satisfying the volume doubling property and generalized Gaussian or sub-Gaussian upper estimates for the heat kernel. We establish its weak type(1, 1) property. In addition, we obtain the weak type(1, 1) of the heat maximal operator in the same setting. Our arguments also work for a large class of direct product manifolds with exponential volume growth. Particularly, we provide a simpler proof of weak type(1, 1) boundedness of some operators considered in the work of Li et al.(2016).

Anesthetic safety evaluation of Enflurane on pregnant dogs and their fetuses

The safety and feasibility of anesthesia during pregnancy are the public concern to human and veterinary medicines,respiratory and cardiac suppression is the most serious side effect of anesthetic. In order to evaluate the effects of Enflurane on pregnant dogs their fetuses,we established the animal model of the artery catheterization,and investigated the maternal and fetal anesthetic concentrations as well as blood gases in the pregnant dogs anesthetized by inhalation of Enflurane. In this experiment,the femoral artery catheterization of 12 pregnant dogs and their 12 fetuses were performed through the anterior tibia artery by using 17 G and 18 G arterial puncture needle,4F expanded tube,3 F induction tube and the C-arm monitoring system. The results showed that the fetuses survived in uterus for 36 h (two cases),4 ~ 5 d (two cases),6 ~7 d (three cases),8 ~10 d (three cases) and 12 ~14 d (two cases) after operation,respectively. 10 fetuses that survived over 4 days in uterus after the operation were successfully delivered. The fetuses were healthy and their catheters were kept well,the other two pregnant dogs aborted their fetuses. After establishing the model successfully,Atropine sulfate in dose of 0. 04 mg/kg bw were injected into cutaneous muscle of dogs in right clinostatism. 15 minutes later after application of Atropine sulfate,the induced anesthesia was carried out by vein injection of 2. 5% Pentothal sodium (10 mg/kg/bw). The tracheal cannula was performed and the dogs were anesthetized for 10 min by Enflurane inhalation from anesthetic machine. Subsequently,the respiratory rate of experimental animals was controlled to 20-22 times/min by a mechanical ventilator,and the tidal volume was 13 ~ 15 mL/kg/bw. The maternal and fetal arterial anesthetic concentrations and blood gases including pH,PO 2,PCO 2,TCO 2,HCO 3-,SB,BEb and Sat. O 2 were measured after Enflurane inhalation anesthesia. The results showed Enflurane can cross placenta and reach fetal blood. The arterial anesthetic concentrations in fetuses were lower than that in their mothers,but the changes of anesthetic concentrations in fetuses were very similar to their mothers. The pH and BEb in the maternal and fetal blood were significantly lower (P 【 0. 01 or 0. 05),PO 2,PCO 2,and Sat. O 2 were significantly higher (P 【 0. 01 or 0. 05),compared to the prior anesthesia. SB decreased,HCO 3-and T-CO 2 increased (P 】 0. 05) in the early stage of anesthesia,but there were no significant difference after and before anesthesia. These results indicated that Enflurane crosses the placenta very quickly in dogs and is characterized by the quick induction anesthesia and elimination in the body. There were slight respiratory and metabolic acidosis in the period of anesthesia in both pregnant dogs and their fetus and the state were gradually restored to normal with the elimination of the Enflurane in blood. Taken together,Enfluence should be considerated the candidate for the induction and maintenance anesthesia for veterinary and human medicine.