Twelve compounds were isolated from the venom of Bufo bufo gargarizans. On the basis of their physical and chemical properties and spectral data, their structures were identified as resibufagenin (1), bufotalin (2), desacetylcinobufagin (3), 19-oxodesacetylcinobufotalin (4), cinobufotalin (5), 1beta-hydroxylbufalin (6), 12alpha-hydroxybufalin (7), bufotalinin (8), Hellebrigenin (9), telocinobufagin (10), hellebrigenol (11) and cinobufagin-3-hemisuberate methyl ester (12), respectively. Compounds 7 and 12 are new natural products.
Animals ; Bufanolides ; chemistry ; Bufo bufo ; Medicine, Chinese Traditional ; Molecular Structure ; Venoms ; chemistry

The safety of Chinese patent medicine has become a focus of social. It is necessary to carry out work on post-marketing clinical safety evaluation for Chinese patent medicine. However, there have no criterions to guide the related research, it is urgent to set up a model and method to guide the practice for related research. According to a series of clinical research, we put forward some views, which contained clear and definite the objective and content of clinical safety evaluation, the work flow should be determined, make a list of items for safety evaluation project, and put forward the three level classification of risk control. We set up a model of post-marketing clinical safety evaluation for Chinese patent medicine. Based this model, the list of items can be used for ranking medicine risks, and then take steps for different risks, aims to lower the app:ds:risksrisk level. At last, the medicine can be managed by five steps in sequence. The five steps are, collect risk signal, risk recognition, risk assessment, risk management, and aftereffect assessment. We hope to provide new ideas for the future research.
Clinical Trials as Topic ; Drug-Related Side Effects and Adverse Reactions ; epidemiology ; etiology ; Drugs, Chinese Herbal ; adverse effects ; chemistry ; economics ; therapeutic use ; Herbal Medicine ; economics ; legislation & jurisprudence ; Humans ; Patents as Topic ; Product Surveillance, Postmarketing ; Quality Control

OBJECTIVETo evaluate the changes in programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) expression on peripheral blood T lymphocytes of patients with chronic hepatitis C (CHC) over the 24 weeks course of antiviral therapy.

METHODSTwenty-four CHC patients administered 24 weeks of combination antiviral therapy with pegylated-interferon-alpha-2a (Peg-IFNa-2a) and ribavirin (RBV) were enrolled for study from the Nanjing Second Hospital between October 2008 and October 2011. Peripheral blood was collected before treatment initiation, at treatment weeks 4, 12 and 24, and post-treatment week 24 (to investigate sustained virologic response (SVR), and used to measure expression of PD-1 and PD-L1 on CD4+ and CD8+ T lymphocytes by flow cytometry, load of serum hepatitis C virus (HCV) RNA by real-time polymerase chain reaction, and level of serum alanine aminotransferase (ALT) by auto-biochemical analyzer. Intergroup differences were analyzed by the two-sample t-test, and the significance of differences between pre- and post-treatment measurements was determined by one-way or two-way repeated measurements analysis of variance tests.

RESULTSAt treatment week 4, 19 of the CHC patients were HCV RNA-negative. Among those patients the PD-1 expression on both T lymphocyte subsets showed a significant decrease from pre-treatment to post-treatment week 24 (CD4+: 18.6 +/- 6.1% vs. 10.3 +/- 7.7%, F = 12.406, P = 0.002; CD8+: 16.6 +/- 13.8% vs. 9.4 +/- 4.6%, F = 4.955, P = 0.039). However, the CD8+ lymphocyte subset showed significant increase in PD-L1 expression during treatment (pre-treatment: 17.5 +/- 13.7% vs. treatment week 4: 25.9 +/- 11.1%, F = 9.063, P less than 0.01; 12: 29.6 +/- 15.1%, F = 8.365, P less than 0.01; 24: 32.0 +/- 15.7%, F = 9.736, P less than 0.01). Among the five CHC patients showing HCV RNA-positivity at treatment week 4 there was only a significant difference observed in the increased expression of PD-L1 on CD8+ lymphocyte subset from pre-treatment to treatment week 24 (17.4 +/- 16.7% vs. 39.2 +/- 15.6%, F = 10.292, P = 0.033). Twenty of the CHC patients achieved SVR. among whom the PD-1 expression was significantly decreased during treatment on the CD4+ lymphocyte subset (pre-treatment: 20.2 +/- 7.5% vs. treatment week 4: 14.4 +/- 7.5%, F = 6.133, P less than 0.05; 12: 14.0 +/- 6.9%, F = 5.541, P less than 0.05; 24: 10.7 +/- 7.6%, F = 14.780, P less than 0.05) and on the CD8+ lymphocyte subset (pre-treatment: 16.8 +/- 13.4% vs. treatment week 12: 10.2 +/- 4.6%, F = 4.964, P less than 0.05; 24: 10.1 +/- 4.9%, F = 4.613, P less than 0.05). Additionally, the PD-L1 expression was significantly increased during treatment on the CD8+ lymphocyte subset (pre-treatment: 19.0 +/- 14.5% vs. treatment week 12: 30.8 +/- 16.6%, F = 6.442, P = 0.020; 24: 35.2 +/- 16.5%, F = 12.349, P = 0.002). Among the four CHC patients who relapsed there were no significant differences observed in the expressions of PD-1 or PD-L1 on the CD4+ or CD8+ T lymphocytes.

CONCLUSIONThe standard Peg-IFNa-2a + RBV combination antiviral therapy reduces PD-1 expression on CD4+ and CD8+ T lymphocytes and increases PD-L1 expression on CD8+ T lymphocytes in peripheral blood. The clinical outcome of CHC patients may be related to the antiviral therapy-induced changes in expressions of PD-1 and PD-L1 on T lymphocytes.

Antiviral Agents ; therapeutic use ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Real-Time Polymerase Chain Reaction ; Ribavirin ; therapeutic use

Objective To study the clinical application anatomy of interventricular foramen and offer a base for operation.Methods Interventricular foramens were observed in 15 adult cadaveric brainThirty- two patients of obstructive hydrocephalus were operated to observe the structure of interventricular foramen un- der neuroendoscope.Results Interventricular foramen was a poriform structure which consists of fornixan- terior pole of thalamencephalon and choroid plexus and was a oval shape in most of themThe plane of the fo- ramen was a included angle with the median sagittal planeThe septal veinthalamostriate veinthalamen- cephal and even the floor of third ventricle could be observed clearly in endoscope.At the same timewe found the foramen had a significant change in obstructive hydrocephalus.Conclusion The interventricular foramen has a simple relatively structure but a variation on size and shape especially in obstructive hydroceph- alusA clearly comprehension of it's structure and adjacent is a base to microsurgery and endoscopic surgery on the foramen.

Objective To investigate the dynamics and development trends of drinking water type of endemic fluorosis after water improvement in Xinbaerhuyouqi of Hulunbeir city, Inner Mongolia and to provide a scientific evidence for the development of countermeasures. Methods We mainly selected Adunchulusumu and Kerlunsumu in Xinbaerhuyouqi of Hulunbeir city as the two monitoring points after water improvement in 2000 -2009. Of these, 1 sample of centralized water supply source water and 3 samples of tap water and 5 samples of noncentralized water supply source water according to water well locations of east, west, south, north and center were collected and the levels of water fluoride were tested; the prevalence of dental fluorosis of school children aged 8 to 12 were examined; from 2002 onwards, the urine samples of 30 children aged 8 to 12(five age groups, six urine samples for each age group) were collected, and all urine samples were collected in the case of less than 30, and urine fluoride was tested. Dental fluorosis was diagnosed using Dean method; water fluoride was tested using fluoride ion selective electrode(WS/T 106-1999); urinary fluoride was tested by determination of fluoride in urine using ion-selective electrode(WS/T 89-1996). Results In 2000 - 2009, the mean levels of fluorine in drinking water in Adunchulusumu and Kerlunsumu were 1.79 - 4.35 mg/L and 1.38 - 3.18 mg/L, respectively; the detection rate of dental fluorosis of children aged 8 to 12 were 45.24％(19/42) - 89.78％(123/137) and 40.00％ (28/70) - 74.47％ (70/94), respectively; the median urinary fluoride of them were 2.30 - 4.15 mg/L and 2.73 - 4.55 mg/L, respectively. ConclusionsThe detection rate of children's dental fluorosis remains high in Xinbaerhuyouqi during the past 10 years after changing water. The endemic fluorosis remains a serious disease. Effective prevention and control measures must be taken to control the occurrence of fluorosis in the future.

Objective To assess the feasibility and efficacy of curved cutter stapler(Contour TM)in total mesorectal excixion with ultra low rectal cancer anal anus-preserving operation.Methods From January to October 2006，36 patients with ultra low rectal cancer fulfilled by anal preserving operation underwent total mesorectal excision with Contour TM. Results Fecal continences were preserved successfully in all patients.Postoperative biopsy of cutting margin was proved to be free of rectal malignant tumors. No anastomotic leakage or narrow and postoperative recurrences occurred in all patients. Conclusion Ultra low rectal cancer resection with Contour is a feasible， safe and convenient technique，which extends the indication range of anal sphincter preservation.

OBJECTIVE To observe the influence of fire-needle therapy on JNK and p38MAPK in ankle-joint of rheumatoid arthritis(RA) rats to explore its underlying mechanism in RA treatment.METHODS 40 male SD rats were randomly divided into normal group,model group,fire-needle group and medication(methotrexate,MTX)group,with 10 cases in each group. Fire-needle was applied to Jiaji(EX-B2) and Ashi point every other day,twice altogether.The right hindpaw swelling volume of rats were detected on the 1st,2nd,4th,6th and 8th day,respectively.The pathological changes in synovial tissue of the right ankle joint were observed by optical microscope.The expression of JNK and p38 was detected by immunohistochemical method and Western blot.RESULTS In comparison with the control group,the swelling paw volumes at the 2nd day signifi-cantly increased of the rest 3 groups(P <0.01).Compared to the model group,the swelling paw volumes on the 8th day in fire-needle group decreased significantly(P <0.05).Serum TNF-αand IL-1β(P <0.05)contents increased in model group compa-ring with normal group,while decreased in fire-needle group and medication group comparing with model group.The patho-logical states of synovial tissue of the right ankle joint in both fire-needle and medication groups were relatively lighter than those of the model group.In comparison with normal group,the expression of JNK and p38 in local synovial tissue of the right ankle joint increased significantly in model group(P <0.05~0.01).Compared with model group,the expression of JNK and p38 of fire-needle and medication group both decreased significantly(P <0.05).These changes were also found in photographs made by immunohistochemical method.CONCLUSION Fire-needle therapy can dowm-regulate the expression of JNK and p38 in local synovial tissue of the right ankle join in MAPK signal transduction pathways,which is the possible mechanism of fire-needle therapy for RA treatment.

Objective To investigate fatigue in children with acute lymphoblastic leukemia (ALL) during chemotherapy,and influencing factors.Methods From Feburary 2014 to August 2016, 216 children with ALL,from 5 to 15 years old,treated in a Grade III,Level A children's hospital in Zhengzhou,were surveyed by Peds QLTM multidimensional fatigue scale (Chinese version) and Questionnaire on discomfort in ALL children during chemotherapy.Results During chemotherapy,total scores of fatigue in the ALL children were (31.71±16.66) ,with fatigue of the whole body (11.67±6.29),of sleeping (10.99±5.57),and of cognition (9.10±6.84);during chemotherapy,incidence of fatigue of the whole body was 90.74%,of sleeping 95.83%, and of cognition 81.94%. Main influencing factors of fatigue of the whole body were weight loss,appetite loss and lack of energy,accounting for 72.8% of fatigue variation;that of sleeping were weight loss and irritability, accounting for 51.2% of fatigue variation;and that of cognition were difficulty in concentration and oral ulcer, accounting for 44.2% of fatigue variation.Conclusions During chemotherapy,fatigue in ALL children is common and is below average level. When managing fatigue symptoms in ALL children,nurses should actively treat the symptoms,so as to reduce the level of fatigue.

Objective To assess the feasibility and efficacy of curved cutter stapler(Contour TM)in total mesorectal excixion with ultra low rectal cancer anal anus-preserving operation.Methods From January to October 2006，36 patients with ultra low rectal cancer fulfilled by anal preserving operation underwent total mesorectal excision with Contour TM. Results Fecal continences were preserved successfully in all patients.Postoperative biopsy of cutting margin was proved to be free of rectal malignant tumors. No anastomotic leakage or narrow and postoperative recurrences occurred in all patients. Conclusion Ultra low rectal cancer resection with Contour is a feasible， safe and convenient technique，which extends the indication range of anal sphincter preservation.

In this study, the molecular marker technology of SRAP and ISSR were applied in rapid identification of seeds from eight species of Brassica oleracea L. Firstly, using the genomic DNA of cabbage as template, SRAP and ISSR reaction systems were optimized through testing every factor, respectively, that affects PCR amplification. Then, using the optimized reaction systems, 30 SRAP primer pairs and 15 ISSR primers were applied to amplify genomic DNA of cabbage, savoy, purple cabbage, borecole, cauliflower, broccoli, Brussels sprouts, and kohlrabi The results showed that high polymorphisms were exhibited among the eight species of Brassica oleracea L. by SRAP primer pairs of M3-E5 and M4-E5, as well as ISSR primers of 844 and 888, especially primer 844 which can identify all eight materials efficiently.
Brassica ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Polymorphism, Genetic ; Repetitive Sequences, Nucleic Acid ; Seeds ; genetics