Objective To optimize the method for total DNA extraction and RAPD analysis of Nervilia fordii (NF),and to study the genetic diversity of different breeds of NF,the substitute of NF and the fake of NF at molecular level.Methods We used low-pH extraction medium with high salt to extract total DNA,and used randomly amplified polymorphic DNAs (RAPD) to select polymorphism primer from 49 random primers.Twenty-two kinds of NF samples were analyzed by RAPD,and classified by SPSS.Genetic diversity were estimated by Shonnon's index and Nei's index.Results A higher puritiy of DNA can be obtained from fresh HF than that from medical materials.We selected 19 polymorphism primers for the cluster analysis of fresh NF and dried medical material.For medical materials of NF,the distance of amplification band of small-leave breed is close to that of middle-size leave breed,but is far away from that of,big-leave breed and Nervilia plicata.For the fresh materials of NF,there breeds of Nervilia fordii can be classified into one kind,the distance of fresh NF band is far from that of Nervilia plicata,Pachyrhizus erosus and the cultured breed,and more far awary from that of Plantago asiatica and Centella asiatica.Shonnon's genetic diversity is 0.463,Nei's genetic diversity is 0.267.Intra-population genetic variation is obvious compared to iner-population genetic variation.The estimated gene flow from Gst (Nm) is 0.94.Conclusion The molecular difference in different breeds of Nervilia Fordii can be used to identify Nervilia Fordii.The genetic diversity of Nervilia fordii is mainly caused by the geography environment.

Animals ; Brain ; metabolism ; Brain Ischemia ; blood ; enzymology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Nitric Oxide ; blood ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

The relationship between acupuncture and stress is discussed from three aspects, including is it possible for acupuncture stimulation to be a stressor, whether acupuncture will start stress reaction, and whether acupuncture effects contain some stress factors. It is believed that correct acupuncture manipulation will not cause stress response, however, under some circumstances, such as inaccurate manipulation, improper treatment or patients who are very nervous but do not receive effective intervention, acupuncture is likely to cause stress response. Acupuncture-induced stress response is totally different from acupuncture anti-stress. The possible stress factors in acupuncture effect are explored, which can provide a new angle for the research on action mechanism of acupuncture. From the view of stress to review acupuncture treatment, there are three enlightenments: emphasizing on communication between doctors and patients, avoiding over-pursuit of deqi and focusing on analysis of the body constitution.
Acupuncture Points ; Acupuncture Therapy ; adverse effects ; psychology ; Humans ; Qi ; Sensation ; Stress, Physiological

Adenocarcinoma ; drug therapy ; immunology ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; drug therapy ; immunology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Killer Cells, Natural ; immunology ; Lung Neoplasms ; drug therapy ; immunology ; Male ; Middle Aged ; Phytotherapy ; Quality of Life ; T-Lymphocyte Subsets ; immunology

Objective To study the changes of mesenteric arterioles on the contractile response to norepinephrine in patients with cirrhosis and portal hypertension (PHT) caused by hepatitis B after treatment by high-dose vitamin C.Methods Eleven PHT patients and 6 non-PHT volunteer were those undergoing their respective surgeriesat in our hospital from January 2010 to June 2011.The PHT patients were randomly assigned to vitamin C treatment group and placebo treatment group.After admission and before induction of anesthesia,blood taken to mesure plasma concentration of malonaldehyde (MDA).Intraoperative portal pressure was measured.We took the third level intestinal mesentery vessels with their mesentery next to the jejunum and studied the contractile response to norepinephrine of the intestinal mesenteric arterioles.We also measured RhoA/ROCK pathway associated proteins' expression and activity changes by Western-blot method.Results The average value of the plasma MDA of PHT patients with cirrhosis was much higher than that of non-PHT patients.After treatment by vitamin C,the value decreased.The dose-response curve of isolated mesenteric arterioles to norepinephrine from PHT patients with cirrhosis shifted to the right,and the EC50 value was higher than that of non-PHT patients.But this phenomenon was reversed after the patients treated with vitamin C.The protein expression of RhoA in mesenteric arteries did not change significantly among the three groups.But the protein expression and activity of ROCK-1 was significantly reduced in PHT patients with cirrhosis.After treatment with vitamin C,the expression and activity of ROCK-1 in mesenteric arteries of cirrhosis was significantly increased.Conclusion The PHT patients with cirrhosis are in the situation of oxidative stress.The high-dose vitamin C treatment can reduce oxidative stress level and improve contractile hyporeactivity of mesenteric arterioles of PHT patients with cirrhosis to norepinephrine.The protein expression and activity increasement of ROCK in RhoA/ ROCK pathway is involved in thisphenomenon.

AIM: To study the therapeutic effect of Angeng Recipe(AGR) on the female climacteric rats. METHODS: Female SD rats were collected and randomly divided into climacterium,AGR(low dose,medium dose and high dose),nilestriol and young control groups.The level of hormone in serum and hypothalamus was analyzed after using AGR,the alteration of organ coefficient of uterus,adrenal,appendix cerebri and thymus were detected,the apoptosis of ovary granulosa cells was detected by fluorescence microscope and flow cytometer. RESULTS: E_2 contents in high dose group were higher than those in the climacterical group.FSH contents in high dose group were lower than those in the climacterical group.Uterus and adrenal gland viscera coefficient of high dose group,medium dose and low dose group were higher than that of climacterical group,The apoptosis of cells in medium dose group showed lower than that in the climacterical group. CONCLUSION: AGR could adjust the function of endocrine secretion by the effect on ovaries and appendices suprasphenoidalis directly or indirectly,and the mechanism of action was multitarget and the regulation was entirety.AGR could relieve the degeneration of sexual gland axis,and improve symptomatic in climacteric.

BACKGROUND:Cuttlebone/racemic polylactic acid composite artificial bone has been prepared in the previous studies to improve the incomplete degradation of cuttlebone. OBJECTIVE:To observe the degradation and biocompatibility of cuttlebone/racemic polylactic acid composite artificial bone in animals. METHODS:Thirty healthy New Zealand white rabbits were randomly divided into four groups. Models of right radial defects were prepared in rabbits, and model rabbits were subjected to implantation of cuttlebone/racemic polylactic acid composite artificial bone into the defects and muscular sac between the radial lateralis muscle and rectus (experimental group), implantation of cuttlebone into the defects and muscular sac between the radial lateralis muscle and rectus (control group 1), implantation of racemic polylactic acid into the defects and muscular sac between the radial lateralis muscle and rectus (control group 2), or no treatment (blank control group), respectively. At 2, 4, 8 weeks after operation, X-ray and histological examinations were performed in the four groups. RESULTS AND CONCLUSION:(1) Compared with the other three groups, the bone mineral density of the experimental group was significantly higher at 4 and 8 weeks after material implantation into the defects (P < 0.05), and moreover, the bone mineral apposition rate of the experimental group was significantly higher at different time after operation (P< 0.05). At 8 weeks after operation, the bone tissues in the experimental group grew from the both ends to the center to form multiple bone island-like structures, with less residual materials, and the marrow cavity and implanting material were in a traffic manner; in the control group 1, there were many residual materials, and no intercommunication was found between the marrow cavity and implant material. (2) At 2 weeks after material implantation into the muscle capsule, there were more inflammatory cels, but the inflammation relieved at 4 weeks and disappeared basicaly at 8 weeks, and the material was degraded partialy. These findings indicate that the cuttlebone/racemic polylactic acid composite artificial bone is a kind of good bone substitute material that has good biocompatibility and degradability.

To give the considerations on the syndrome differentiation and acupuncture treatment of diabetic peripheral neuropathy, generalize its pathogenesis and summarize treatment rules, this paper reviews and analyzed 52 articles on acupuncture treatment in recent years. It is believed that diabetic peripheral neuropathy is rooted from qi-yin deficiency and presented with blood stasis and collateral obstruction; many organs are affected, the liver, kidney,spleen and stomach in particular; the treatment principle concentrates on unblocking collaterals and regulating organs; treatment centers on limb acupoints, Back-Shu point and Front-Mu point;acupuncture treatment should be adopted early as possible and its clinical efficacy is rather good.

Objective To analyze the bibliometry of the literature of acupoint thread-embedding in treating simple obesity, and to provide references for clinic.Method By both computer and manual retrieval, the relevant literatures over the past 30 years were collected from the Chinese Journal Full-text Database (CJFD), Vip database, WanFang database. By using the SPSS 17.0 software for statistical analysis, the distribution (province or city), publication year, and journal of the literatures, supporting fund, sample size, research method, material of the thread, use frequency of the involved acupoints, and the meridians of the selected acupoints were summarized.Result The included eligible 175 articles were distributed over many provinces and cities; the publication amount had been increasing year by year; there was a variety of thread materials; the literature with supporting funds only occupied a small percentage, while the research with a small sample size occupied a comparatively large percentage, and the randomized controlled trials (RCTs) also had a small percentage.Conclusion As a common Chinese external therapy in treating simple obesity, acupoint thread-embedding has been paid more and more attention for its practical value and bright future. However, the relevant governmental department hasn’t paid enough attention to this therapy, and the low clinical research quality also makes it difficult to scientifically evaluate its clinical efficacy and safety. Therefore, more attention is required, RCTs of multicenter and large sample size are expected, and clinical research quality also needs improvement, to provide references for clinic.

Objective To optimize the sulfated modification conditions for Rhizoma Smilacis Glabrae polysaccharides (RSGP),and to investigate the possibility of enhancing the activity of RSGP after sulfating modification.Methods RSGP was modified by cholorosulfonic acid-pyridine.With the yield,carbohydrate content and sulphate substitution degree as the observation indexes,L9 (34)orthogonal design was used to optimize reaction time,reaction temperature and reagent ratio.The degree of sulphate substitution was determined by barium chloride turbitimetry,and the carbohydrate content was detected by sulfuric acid-phenol method.Then the structures of the sulfated modified products were analyzed by infrared radiation (IR) spectrum.Methyl thiazolyl tetrazolim(MTT)assay was used to determine the anti-tumor activity of sulfated RSGP on HepG2 and MCF-7 cell lines.Results The optimized modification conditions of RSGP were sulfating RSGP with cholorosulfonic acid-pyridine in the ratio of 1:6 for 4.5 hours at 60 ℃.The MTT assay results showed that the sulfated RSGP could inhibit the growth and proliferation of human liver cancer HepG2 cell line and human mammary cancer MCF-7 cell line in concentrationdepending manner.Conclusion It is feasible for sulfating modification of RSGP with cholorosulfonic acid-pyridine,and sulfating modification can enhance the anti-tumor activitv of RSGP.