Objective To study the genetic diversity of Fritillaria thunbergii,a traditional Chinese herb in Zhejiang Province in China.Methods The genetic diversity of six representational populations of F.thunbergii including 32 individuals was investigated by amplified fragment length polymorphism(AFLP) maker technique.Results The genetic diversity was revealed as follow: the Nei′s genetic diversity index(He) 0.169 0?0.175 7,Shannon′s information index(I) 0.269 8?0.245 3,percentage of polymorphic loci(PPB) was 76.85% at the species level;Ht 0.169 0?0.030 9,and Hs 0.150 8?0.024 0,I 0.233 3?0.261 9, PPB was 50.38% at population level.The genetic differentiation index(Gst) was 0.107 6,Nm 4.147 0.The result of dendrogram of six populations indicated that Dongyang and Yongkang populations shared the minimum genetic distance(0.015 0),they were classified into a group,and Xiangshan and Jinyun populations shared the maximum genetic distance(0.032 4).Conclusion The genetic diversity of F.thunbergii cultivated in Zhejiang Province is very rich,which could ensure the long-term survival of F.thunbergii.But the genetic diversity of F.thunbergii is relatively higher in population levels while lower at the species levels and the degree of genetic differentiation occured among the populations is not significant.The germplasm resources are relatively stable among these six populations.These populations could be used to breed the fine strains of F.thunbergii as the bases.

Objective To observe the genetic structure of cultivated Scrophularia ningpoensis in Zhejiang Province.Methods The genetic structures of six typical S.ningpoensis populations were analyzed by fluorescence AFLP marker.Results Bands(12 552) were generated by seven pairs of AFLP primer combinations,of which 8 808 were polymorphic,and the polymorphic rate was 70.17%.The variety ranges of PPB among different populations were 41.67%—55.56%,and 47.30% in average.I was between 0.190 8—0.238 3,and 0.221 8 in average.Ne was between 1.201 4—1.280 6,and 1.236 9 in average.Gst was 0.127 1,Nm was 3.432 4.UPGMA Cluster analysis showed that the six populations can be divided into two clusters,as that of Tiantai,Jinyun,and Jingning were one sub-cluster,and Dongyang,Pan′an,and Xianju were another one sub-cluster.Conclusion There is a relative high genetic diversity level in cultured S.ningpoensis of Zhejiang Province.Genetic differentiation exists among populations,but it exists in population mostly.There is a relative high genetic intercommunion among populations.The genetic distance is not related to the geographic environment.

Objective To study the association between human papillomavirus (HPV) genotypes and viral load and clinical features of verruca vulgaris.Methods Tissue samples were collected from 48 outpatients with verruca vulgaris,and DNA was extracted from these tissue samples.To determine the genotype of HPV,PCR was performed to amplify the L1 fragment of HPV with universal primers followed by bidirectional sequencing and BLAST.The genotyping results were validated by PCR with type-specific primers.Real-time fluorescence-based quantitative PCR was conducted to measure the viral load of HPV,and hematoxylin and eosin (HE) staining to observe histological changes in these tissue specimens.Results The L1 fragment of HPV was amplified from 35 out of the 48 tissue specimens.Of the 35 L1-positive specimens,32 harbored HPV 7,1 harbored HPV 57,and 2 harbored both HPV 2 and HPV 7.Multiple lesions were observed on extremities in the patient infected with HPV 57,but on the head,face and trunk in the patients coinfected with HPV 2 and HPV 7.There were no significant differences in HPV viral load or vacuolated cell number between patients with single lesions and those with multiple lesions,or between patients with a clinical course of ＜ 6 months and those with a clinical course of 6-12 months.However,HPV viral load tended to decrease one year after the onset,and there was pronounced hyperkeratosis and less vacuolated cells in lesions of long duration (more than 2 years) compared with those of short duration (less than 2 years).Conclusions HPV 7 appears to be the most common HPV genotype associated with verruca vulgaris,and HPV 7 infection usually occurs on the head and face.For verruca vulgaris of less than 1 year,neither HPV viral load nor vacuolated cell number is associated with the count or clinical course of warts.

Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

OBJECTIVETo study the triterpenoid saponins in Hemsleya chensnsis.

METHODThe chemical constituents were isolated and purified by various chromatographic methods and their structures were elucidated on the basis of spectral analysis.

RESULTSeven known triterpenoid saponins were isolated from the root of H. chensnsis and were identified as 3-O-beta-D-glucuropyranosyl-oleanol-icacid (1), 3-O-beta-D-glucopyra-noside-oleanolicacid-28-O-beta-D-glucopyranoside (2), 3-O-(6'-methylester)-beta-D-glucuropyranosyl olea- nolic acid-28-O-alpha-L-arabinopyranoside (3), 3-O-(6'-methylester)-beta-D- glucuropyranosyl- oleanolic acid-28-O-beta-D-mannupyranoside (4), 3-O-(6'-ethyl ester)-beta-D-glucuropyranosyl oleanolic acid-28-O-beta-D- glucopyranoside (5), 3-O-alpha-L-arabinopyranoside-(1-->3) -beta-D-glucuropyranosyl- oleanolic acid-28-O-beta-D-glucopyranoside (6), 3-O-beta-D-glucu-ropyranosyl-oleanolic acid-28-O-beta-D-glucopyra-noside-(1-->6)-beta-D-glucopyranoside (7).

CONCLUSIONCompounds 1-7 were isolated from this plant for the first time.

Cucurbitaceae ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Mitragyna ; chemistry ; Molecular Structure ; Plant Extracts ; pharmacology ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

Objective To investigate the possible risk factors for the progression of abdominal aortic calcification (AAC) in MHD patients.Methods Total of 170 patients on MHD between June 2014 and October 2014 in the dialysis center of the Second Hospital of Tianjin Medical University were included prospectively.Lateral lumbar radiography were applied to evaluate patients' AAC score (AACs) at baseline and after two-years of follow-up respectively.According to the change of AACs,the patients were divided into rapid AAC progression group and non-rapid AAC progression group.Multivariable Logistic regression models were used to determine the risk factors for the progression of AAC in MHD patients.Results At baseline,the presence of AAC (AACs≥1) was 43.5％(74/170).The mean follow-up duration was 27.6(24.7,28.0) months.AACs were available in 111 patients,and the presence of AAC was 78.4％(87/111).During the follow up,36 patients developed new AAC;rapid AAC progression was seen in 54 patients,and non-rapid AAC progression was seen in 57 patients.Multivariate Logistic regression analysis demonstrated that hyperphosphatemia (OR=4.373,95％ CI 1.562-7.246,P=0.005) and high density lipoprotein (HDL) (OR=0.031,95％CI 0.003-0.338,P=0.004) were independent risk factors for AAC progression in MHD patients.Conclusions Hyperphosphatemia and low HDL may promote the progression of AAC.Well-controlled serum phosphate and lipid metabolism may slow the progression of vascular calcification,reducing cardiovascular morbidity and mortality.

Objective To investigate the effects and related risk factors of different vascular access types on new atrial fibrillation in maintenance hemodialysis (MHD) patients.Methods This was a single-center prospective cohort study.Patients who established long-term dialysis access and were voluntarily followed up in the Second Hospital of Tianjin Medical University from January 1,2013 to June 30,2013 were enrolled to follow-up for 5 years.Patients were divided into fistula group (patients with autogenous arteriovenous fistula) and catheter group (patients with tunneled cuffed internal jugular vein catheter).The incidences of new atrial fibrillation in the two groups were compared by Kaplan-Meier survival analysis.Cox regression analysis and receiver operator characteristic curve (ROC) were used to assess the risk factors of new atrial fibrillation.Results A total of 315 eligible patients were enrolled,including 150 males (47.62％).There were 189 patients (60.00％) in the fistula group,and 126 patients (40.00％) in the catheter group.Multivariate Cox regression analysis showed that older age (HR=1.021,95％CI 1.003-1.040),arteriovenous fistula (HR=1.899,95％CI 1.019-3.539),increased dialysis blood flow (HR=1.030,95％CI 1.010-1.051) and left atrial diameter (HR=1.097,95％CI 1.022-1.177) were independent risk factors for new atrial fibrillation in MHD patients (all P ＜ 0.05).Kaplan-Meier survival analysis showed that the incidence of new atrial fibrillation in fistula group was higher than that in catheter group (Log-rank A2=9.53,P=0.002).ROC curve analysis showed that age [the area under the curve (AUC)=0.608,P=0.008],arteriovenous fistula (AUC=0.594,P=0.021),dialysis blood flow (AUC=0.659,P＜0.001) and left atrial diameter (AUC=0.604,P=0.011) could predict the occurrence of new atrial fibrillation.Condusions Older age,arteriovenous fistula,increased blood flow during dialysis and left atrial diameter are independent risk factors for new atrial fibrillation in MHD patients,which can predict the occurrence of atrial fibrillation.The incidence of new atrial fibrillation in patients with arteriovenous fistula is higher than that in patients with catheter.

One hundred and twenty maintenance hemodialysis patients with secondary hyperparathyroidism were randomly assigned to receive cinacalce,calcitriol or combination of cinacalce and cacitriol for treatment,with 40 cases in each group.Patients were followed up for 12 months;and the blood tests,echocardiography,examinations for osteoporosis and soft tissue calcification were performed every month.After 3 months of treatment,the serum levels of parathyroid hormone were decreased in all three groups (P ＜ 0.01);while the parathyroid hormone decreased more markedly with less influence on serum calcium and phosphorus levels in combination group.There were no significant changes in alkaline phosphatase,osteoporosis and cardiac valve calcification after treatment,compared with before treatment.There were no cases of hypercalcemia and hypocalcemia appearing in combination group after treatment.The study indicates that the combination of cinacalcet with calcitriol has better therapeutic effect for treatment of secondary hyperparathyroidism in maintenance dialysis patients.

Objective To investigate the efficacy and safety of neurotropin in the treatment of restless legs syndrome and sleep disorder in patients with maintenance hemodialysis. Methods Sixty eight patients who met the inclusion criteria were randomly assigned to control group (n=34) and treatment group (n=34). The trial lasted for 16 weeks, and all patients undergone thorough dialysis. 7.2 units (2 branches) neurotropin were slowly injected to the patients in the treatment group at the end of each hemodialysis and they were stopped after 8 weeks. The patients in the control group had no treatment for restless leg syndrome on the basis of adequate dialysis. All patients were assessed regularly as regards their biochemical indexes, restless legs syndrome rating scale and Pittsburgh sleep quality index. Results All the patients completed the experiment, and restless legs syndrome scores were decreased in two groups. Compared with the baseline, the restless legs syndrome scores decreased significantly in patients treated with neurotropin, and the differences between two groups were significant (P<0.01). In the Pittsburgh sleep quality index scores, the patients in the treatment group decreased in all scores and the difference was statistically significant (P<0.01). Conclusions Neurotropin can relieve the symptoms of restless legs syndrome and sleep disturbance in patients on maintenance hemodialysis and is safe, but symptoms may occur again after withdrawal.