Tropane alkaloids,such as hyoscyamine (or its more stable racemate atropine) and scopolamine,are remembered as the oldest drugs in medicine with wide pharmaceutical applications clinically for their mydriatic,antispasmodic,anticholinergic,analgesic and sedative properties.Presently,the supply of tropane alkaloids are entirely rested on their isolation from plant materials,yet the low contents of tropane alkaloids in plants raise the requirement to breed new varieties of high-yield tropane alkaloids and improving the production of tropane alkaloids in hairy root cultures or plants by biotechnology has be a research focus in the field of secondary metabolism.For the past decade,there have been important progresses on biosynthesis of tropane alkaloids and their molecular biology,some new pathway genes have been identified and many achievements have also been acquired in metabolic engineering of tropane alkaloids.This review summarized the recent advances in above aspects,and the problems and orientations of future research are also discussed and proposed.

Objective To observe the effect of topical application with warm and wet carthamus tinctorius alcohol and anisodaminum on phlebitis caused by mannitol injection. Methods 100 patients with fractured bones suffered from phlebitis caused by mannitol injection were randomized into the observation group and the control group with 50 cases in each group. The observation group adopted topical application with warm and wet carthamus tinctorius alcohol and anisodaminum while the control received external application of 50% magnesium sulfate. The dosage was 3 hours per time,2 times a day, one time in the morning and afternoon respectively. The treatment continued till the phlebitis disappeared and the effect at the 24th,48th and 72th post treatment was compared between the two groups. Results The effect at the above mentioned time points in the obsevation group was superior to that of the control group(P＜0.05).Conclusion Topical application with warm and wet carthamus tinctorius alcohol and anisodaminum on phlebitis caused by mannitol injection proved to be safe and had no adverse effect.

Objective To translate the Personal Diabetes Questionnaire(PDQ),and test the relia bility and validity of the Chinese version of PDQ.Methods Through forward translation,back translation,cultural adjustment,etc.,the Personal Diabetes Questionnaire (PDQ) was introduced to localize it.60 eligible patients with diabetes were selected in a tertiary hospital to make a preliminary investigation.The results were statistically analyzed in order to assess the reliability and validity of the Chinese version of PDQ.Results The Chinese version of PDQ had good reliability,whose total internal consistency coefficient was 0.831 and test-retest reliability was 0.801.The questionnaire has good validity.Total Content Validity Index was 0.938.I-CVI was between 0.800 and 1.000.The questionnaire's dimensions had good parallel validity with glycosylated hemoglobin (HbAlc) and body mass index (BMI).Conclusions The Chinese Version of the personal diabetes questionnaire has good reliability and validity,which can be widely used as the evaluation tool of diabetes self-management education.

Objective The objective of this study was to explore the difference factors for PHC patients by transcatheter arterial chemoembolization of different age groups of senior people in China and support reference for individualized intervention by clinical medical workers.Methods 153 cases of PHC patients by TACE were selected by convenience sampling method from October 2013 to June 2014.The data were collected using general data questionnaire,Health Promotion Strategy Scale,QOL-LC,Memorial Symptom Assessment Scales and Social Support Scale to evaluate the physical,psychological and social aspects of the PHC patients.We divided the patients into young,middle-aged and elderly groups.Single factor and multiple factors regression analysis were used to compare the status and influencing factors.Results Psychological function,value of blood AFP,marital status entered the model and were self-care efficacy and correlated factors of young-aged patients,which could explain 50.6％ of the total varience.Physical symptoms,exploitation degree of support and intervention treatment times entered the model and were self-care efficacy and correlated factors of middle-aged patients,which could explain 47.8％ of the total varience.Symptom distress,degree of cuhural and psychological function entered the model and were self-care efficacy and correlated factors of elderly-aged patients,which could explain 42.4％ of the total varience.Conclusions The self-care efficacy were low in all ages of PHC patients by TACE,it is associated with a variety of factors.Doctors and nurses should pay more attention to the self-care efficacy for PHC patients by transcatheter arterial chemoembolization with different age groups.Also we should evaluate and make well-targeted policies and personalized inter-ventions to promote self-management self-efficacy.

Objective To investigate the risk of permanent hair dye use for connective tissue disease ( CTD ) with positive antinuclear antibodies ( ANA ).Methods We conducted a population-based case-control study to analyze 440 CTD patients from the department of our hospital.Four hundreds and forty patients' relatives or friends who were free from any CTD were selected as controls.Patients and controls were matched in sex and age.A face-to-face interview was adopted to finish questionnaires about subjects' demographic data and the use of permanent hair dye.Data about the duration,frequency,times of usage and allergy related to hair dye were collected.Medical history was also reviewed,such as the diagnosis,disease course,primary symptoms,serum tests and organ involvements.Statistical analysis was performed by SPSS 17.0 software using x2 test and Logistic regression.Results The association between the use of permanent hair dye and CTD did not reach statistical significance (OR=1.282,95％CI 0.966-1.700,P=0.085 ),when the relationship of hair dye use with systemic lupus erythematosus (SLE) was tested,the same result (OR=1.092,95％CI 0.795-1.500,P=0.587 ) could be revealed.Conclusion This preliminary study has shown that the use of permanent hair dye couldn't induce the occurrence of CTD.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

AIM:Activin A,a member of transforming growth factor superfamily,is the negative regulator factor in liver regeneration. In this study,the effects of extract of Ginkgo Biloba on hepatic fibrosis and the expression of Activin A in rats with cirrhosis were investigated. METHODS:The experiment was performed at in the Central Laboratory of Wuhan First Hospital from September 2005 to December 2006. ①Thirty-six male SD rats of(160?20) g were randomized into 3 groups:control group,model group and treatment group. ② Except the rats in the control group,others were intraperitoneally injected with 500 mL/L CCl4 for 8 weeks to establish models of hepatic fibrosis. Meanwhile,the extract treatment group was infused with the extract of Ginkgo Biloba(Chinese drugs preparation laboratory of Wuhan First Hospital,detected by Hubeu Wushi Medicine Industry Co.,Ltd. No. 02-391) daily for 8 weeks. ③After administration,all anesthetized rats were sacrificed. Blood samples were collected for the determination of liver function biochemical indexes. Liver tissue samples were used for histopathological examinations. The expression of Activin A was determined by immunohistochemistry and RT-PCR. RESULTS:All 36 rats were involved in the final analysis. ①The liver function in extract treatment group was significantly improved compared with that in model group. ②The grade of fibrosis in extract treatment group were remarkably lower than that in model group under light microscope. ③The positive staining of Activin A in treatment group was significantly reduced compared with model group. ④The expression of Activin A mRNA in extract treatment group was significantly reduced compared with model group. CONCLUSION:Extract of Ginkgo Biloba can effectively decrease the expression of Activin A in rats with hepatic fibrosis caused by CCl4,and lessen the degree of hepatic fibrosis.

Objective To establish an effective PCR assay for detection of Bartonella, and application of this assay in tree shrew .Methods Sequence of Bartonella was obtained from NCBI Genbank .Three pairs of primers were designed based on this sequence .One pair of primers was determined through amplifying the major strains in China .Sixty tree shrew blood samples were tested with this PCR assay .The positive amplified fragments were sequenced to verify the reliability of this method .Results A PCR method for detection of Bartonella is successfully established , with a high specificity and the sensitivity was of 2.0 ×10 -5 μg/mL.Among the tested 60 blood samples , 15 positive cases were detected.Sequencing of the samples confirmed a 25%infection rate of Bartonella in the tree shrews, well consistent with the amplification results , and verified the applicability of this detection method .Conclusion The establishment of this method provides the basis for detection of Bartonella in tree shrew.

ObjectiveTo investigate the mechanism of anti-inflammatory effect of diallyl sulfide (DAS) in protection against acute lung injury (ALI) in rats with paraquat poisoning.Methods Eighty male Wistar rats were randomly divided into four groups, namely: control group, model group, dexamethasone (DXM) treatment group, and DAS treatment group, with 20 rats in each group. The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by gavage, while the same volume of normal saline (NS) was given in same manner in control group. 100 mg/kg of DAS, the same volume of NS, or 1 mg/kg DXM injection were given respectively in DAS treatment group, model group, or DXM treatment group intraperitoneally after exposure to paraquat, once a day for 14 days. Five rats in each group were sacrificed at 1, 3, 7, 14 days, respectively. The inferior lobe of right lung was harvested, and the degree of lung injury was observed with hematoxylin and eosin (HE) staining under optical microscope; the upper lobe of right lung was used to determine the lung wet/dry weight (W/D) ratio and for evaluation of the degree of pulmonary edema. The expression of nuclear factor -κB (NF-κB) in the middle lobe of right lung was assessed with immunohistochemistry. The expression of tumor necrosis factor -α (TNF-α) mRNA in the left lung was determined with the reverse transcription-polymerase chain reaction (RT-PCR).Results① The pulmonary structure in control group was found to be intact. However, in the model group there were progressive pathological changes in lung, including marked edema and thickening of alveolar walls, collapse of alveoli, infiltration of inflammatory cells, alveolar wall, and obvious bleeding in the local lung tissue, and formation of transparent membrane in alveolar space. Less infiltration of inflammatory cells and no obvious destruction were found in alveolar structure in the DAS and DXM treatment groups.② Lung W/D ratio: lung W/D ratio of model group was apparently higher than that in control group at every time point, and peaking on the 3rd day (6.15±0.54 vs. 4.15±2.10,P< 0.05), and the ratio of lung W/D of DAS and DXM treatment groups was obviously lower than that in model group at every time point, especially on the 3rd day (3.99±1.26, 4.30±0.70 vs. 6.15±0.54, bothP< 0.05), but there was no significant difference between DAS and DXM treatment groups in this regard.③ The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell nuclei of the control group, while extensive NF-κBp65 expression was found in model group. Minimal NF-κBp65 positive expression in the cytoplasm and even less positive expression in the nucleus was found in the DAS and DXM treatment groups, and integralA value was significantly lower in the DAS and DXM treatment groups than that of the model group, especially on the 3rd day [(17.98±0.06)×107, (18.53±0.04)×107 vs. (28.85±0.61)×107, bothP< 0.01], but there was no significant difference between DAS and DXM treatment groups.④ It was shown by RT-PCR that the expression of TNF-α mRNA in lung tissue of the model group was significantly higher than that in the control group on the 3rd day (gray value: 3.63±0.62 vs. 0.51±0.13, P< 0.05). The expression of TNF-α mRNA in lung tissue was significantly decreased in DAS and DXM treatment groups compared with model group (gray value: 2.49±0.57, 2.02±0.26 vs. 3.63±0.62, bothP< 0.05), but there was no significant difference between DAS and DXM treated groups.ConclusionTreatment with an intraperitoneally injection of DAS is capable of attenuate the extent of PQ-induced ALI in rats by alleviating pulmonary edema, inhibiting the expression of NF-κB and TNF-α in lung tissue, and ameliorating pathological changes in lung tissue.

Objective To investigate the protective effects of rhein lysinate (RHL)on the blood vessel damage induced by oxidative stress in the mice,and to explore its mechanism.Methods The mouse models of oxidative damage were established by intraperitoneal injection of paraquat.30 C57 mice were randomly divided into control, paraquat model,and RHL prevention groups.The mice in RHL prevention group were given RHL by gavage for one week before performing model.The mice in other two groups were given equal volume of distilled water.For making model,paraquat was intraperitoneally injected in the mice in paraquat model and RHL prevention groups once a week for two weeks.The activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) and the content of serum malonaldehyde (MDA) of the mice were detected 2 weeks after modeling. The pathological profile of blood vessel was observed by hematoxylin and eosin (HE)staining and the level of reactive oxygen species was observed by DCFH-DA staining.The expressions of genes related to blood vessel damage were detected by Western blotting method.Results Compared with control group,the activities of SOD and GSH-Px were decreased and the content of MDA was increased in paraquat model group (P < 0.05 ). Compared with paraquat model group,the activities of SOD and GSH-Px were increased and the content of MDA was decreased in RHL prevention group (P <0.05).The pathological examination indicated the structure of blood vessel of the mice was damaged and the level of reactive oxygen species of blood vessel was increased (P <0.05)in paraquat model group.The pathological changes were significantly improved and the level of reactive oxygen species of blood vessel of the mice was decreased (P < 0.05 )in RHL prevention group. The Western blotting analysis showed that compared with control group,the expression levels of nitric oxide endothelial synthase (eNOS)and caspase-3 of the mice in paraquat model group were decreased (P < 0.05),however the expression level of cleaved fragment of caspase-3 was increased (P < 0.05).Compared with paraquat model group,the expression levels of eNOS and caspase-3 of the mice in RHL prevention group were increased (P < 0.05 )and the expression level of cleaved fragment of caspase-3 was decreased (P <0.05).Conclusion Paraquat could induce vascular cell damage in vivo through increasing the levels of reactive oxygen species, and RHL could antagonize the effects of paraquat by scavenging reactive oxygen species, and up-regulating the eNOS expression and reducing the expression of the cleaved fragment of caspase-3.