In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.
Amplified Fragment Length Polymorphism Analysis ; methods ; Atractylodes ; genetics ; Sequence Analysis, DNA ; Tetraploidy

Objective To determine the impact of fluorine and aluminum,and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro and to explore its mechanisms.Methods The osteoclasts and bone marrow stromal cells (BMSCs) isolated from long bone of new born rats were cultured,respectively,in TC199 medium (containing 10％ fetal bovine serum) with fluoride,aluminum and fluoride combined with aluminum.The osteoclasts were inoculated in 96-well culture plate and ivory slice,BMSCs in 6-well culture plate,and culture medium was changed after 2 hours incubation.The cells were divided into control group,fluoride group,aluminum group and fluoride combined with aluminum group; the doses of sodium fluoride were 0,1.0 × 10-4,0,1.0 × 10-4 mol/L and the doses of aluminum chloride were 0,0,1.0 × 10-5,1.0 × 10-5 mol/L,respectively.Tartrate-resistant acid phosphatase (TRAP) staining positive cells were counted under light microscope after TRAP staining on the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue.The expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) was detected by real-time fluorescence quantitative PCR in BMSCs after 8 h treatment.Results ① Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the numbers of osteoclasts (F =7.15,6.56 and 7.98,respectively,all P ＜ 0.05).The numbers of osteoclasts in fluoride group,aluminum group and fluoride combined with aluminum group[(136.9 ± 22.99),(135.4 ± 23.5),(163.0 ± 24.4) per well] were higher than that in the control group[(92.5 ± 22.1) per well,all P ＜ 0.05].② Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the resorption pit area on ivory slices(F =10.47,12.64,14.29,respectively,all P ＜ 0.05).The resorption pit area on ivory slices in fluoride group,aluminum group and fluoride combined with aluminum group[(0.242 ± 0.031),(0.293 ± 0.026),(0.333 ± 0.016)mm2 per slice] was higher than that in the control group [(0.088 ± 0.030)mm2 per slice,all P ＜ 0.05].③Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the expression ratios of RANKL/OPG in BMSCs (F =8.15,15.38,23.59,respectively,all P ＜ 0.05).The expression ratios of RANKL/OPG in BMSCs in fluoride group,aluminum group and fluoride combined with aluminum group [(193.98 ± 137.93)％,(326.11 ± 176.78)％,(599.84 ± 275.82)％] were higher than that in the control group[(100.00 ± 56.02)％,all P ＜ 0.05].Conclusions Both fluoride and aluminum can cause increase in the number of osteoclasts in vitro and promote cell differentiation and bone resorption activity,which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.The stimulating effects of fluoride on osteoclasts differentiation and bone resorption is enhanced by aluminum.

Objective To determine whether arsenic has estrogen-like effects,the cell proliferation was measured iil human eervical cancer line(HeLa)in vitro.Methods The HeLa cells were grown in improved RPMI 1640 supplemented respectively with β-estradiol(E2,1 nmol/L),Arsenic trioxide(As2O3,0.5,1.0,5.0 μmol/L),ICI (500 nmol/L),E2(1 nmol/L)+ICI(500 nmol/L),As2O3(1.0 μmol/L)+ICI(500 nmol/L)and control.The growth morphology of HeLa cell was observed under microscope after 72 h.The method of M1Tr was used to study the cell proliferation after 24.48 and 72 h.The technique of flow eytometry was used to measure cell cycle after 48 h. Results HeLa cells in E2 and 0.5 μmoL/L As2O3 treatment were more better growth in morphology than control group.Percentage of HeLa cells proliferation at 24,48,72 h in E2 and 0.5 μmol/L As2O3 treatment were 6.35%, 11.56%,38.33%and 6.35%,8.50%,20.26%respectively.The proliferation effect of HeLa cells was similar in two treatments.The proliferation of HeLa cells were inhibited in other treatments.Compared with control[(41.68± 1.05)%],HeLa cells were promoted go to S phases in E2[(55.72±2.31)%]and 0.5 μmol/L As2O3[(47.82± 1.41)%]treatment.But in other treatments HeLa cells were hold back to S phases.Compared with control,there was a significant differenee(P＜0.05)of cell percentage in S phases in 5.0 μmol/L As2O3[(21.11±4.99)%]and ICI[(20.16±4.76)%]treatments.Conclusion Small amounts of As2O3 impose estrogen.1ike effects and stimulate the proliferation of HeLa cells.

Objective To determine the effects of fluoride on osteoclasts's quantity and bone resorption function in vitro and its mechanisms. Methods The osteoclasts and bone marrow stromal cells(BMSCs) isolated from long bone of new born rats were cultured respectively in TC199 medium (containing 10% fetal bovine serum) with fluoride. The osteoclasts were inoculated in 96-well culture plate and ivory slice, BMSCs were inoculated in 6- well culture plate, respectively, medium were changed after 2 hours incubation. They were divided into control group, low-dose fluoride, medium-dose fluoride and high-dose fluoride groups, the doses of sodium fluoride were 0,2.5 × 10-5,5.0 × 10-5,10.0 × 10-5 mol/L, respectively. Tartrate-resistant acid phosphatase(TRAP) staining positive cells were counted under light microscope after TRAP staining on the 2nd and the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue. The expression of receptor activator of NK-κβ ligand(RANKL) and osteoprotegerin(OPC) was detected by real-time fluorescence quantitative (337.5 ± 70.5), (447.5 ± 43.4), (472.9 ± 34.8), (475.3 ± 24.3)/well in the control group, the low-dose, mediumdose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the control group (all P < 0.05). After in vitro culture for 5 days, the numbers of osteoclasts were (92.5 ± 22.1), (123.0 ± 26.4), (135.5 ± 22.2), (136.9 ± 23.0) per well in the control group, the low-dose, medium-dose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the (0.088 ± 0.030), (0.100 ± 0.018), (0.152 ± 0.015), (0.242 ± 0.031 )mm2 per piece in the control group, the lowdose, medium-dose and high-dose fluoride groups, respectively. The values of medium-dose and high-dose fluoride BMSCs in the control group, the low-dose, medium-dose and high-dose fluoride groups were 100.00 ± 56.02, 144.95 ± 97.21,223.25 ± 184.48,193.98 ± 137.93, respectively. The values of medium-dose and high-dose fluoride groups were significantly higher than that of control group (all P < 0.05). Conclusions Fluoride can cause increase in the number of osteoclasts in vitro and promote their cell differentiation and bone resorption activity, which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.

Objective To observe the influence of longmuzhuanggu powder(LMZGP)on gastroenteric movement and plasm motilin(MTL)in rats.Methods Rats were randomly divided into control group,model group,LMZGP large dose group,LMZGP middle dose group,and LMZGP low dose group.To establish the model of disorder of gastric depletion and small intestinal advancement by using maxolon,atropine,neostigmine and adrenaline.Gastric emptying and intestinal progradating rates in mice were tested with black nutritious semi solid cataplasm.Level of MTL in hemtatoplasma was tested by radio immunoassay.Results Maxolon accelerated gastric depletion and neostigmine accelerated intestine propagation and heightened the level of MTL in hematoplasma(P_a

Actins ; metabolism ; Adenocarcinoma, Clear Cell ; metabolism ; pathology ; Adult ; Carcinoma, Renal Cell ; metabolism ; pathology ; Desmin ; metabolism ; Diagnosis, Differential ; Endometrial Stromal Tumors ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; Leiomyoma, Epithelioid ; metabolism ; pathology ; Melanoma ; metabolism ; pathology ; Melanoma-Specific Antigens ; metabolism ; Perivascular Epithelioid Cell Neoplasms ; metabolism ; pathology ; surgery ; Receptors, Progesterone ; metabolism ; Uterine Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To investigate the effect of non-alcoholic fatty liver disease (NAFLD) on virologic response in chronic hepatitis B patients treated with nucleos(t)ide analogues.Methods Three hundred and thirty-two treatment-naive patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) who visited clinic or hospitalized in the First Affiliated Hospital of Wenzhou Medical College from January 2007 to December 2009 were enrolled and finally 316 patients completed the research.Among them,106 patients were diagnosed with CHB complicated with NAFLD by ultrasonography (NAFLD group),and the remaining 210 patients were diagnosed with simple CHB (CHB group).Patients in both groups were treated with combination therapy of lamivudine (LAM) plus adefovir dipivoxil (ADV) or entecavir (ETV) monotherapy for 96 weeks.Alanine aminotransferase (ALT) levels,virologic response and serological response were compared pre-and post-treatment in both groups.Enumeration data was compared using x2 test,and correlation analysis was performed using Logistic multivariate regression analysis.Results Rates of hepatitis B virus (HBV) DNA clearance in the NAFLD group at week 12,24 and 48 were 46.2％,61.3％ and 74.5％,respectively,which were all significantly lower than those in CHB group (61.9％,73.8％ and 83.8％,x2 =7.051,5.195,3.895,respectively; all P＜0.05).However,rates of HBV DNA clearance were 85.8％ in the NAFLD group and 91.9％ in the CHB group at week 96.This difference did not reach the level of significance (x2 =2.838,P＞0.05).Rates of HBeAg loss in the NAFLD group were 27.4％ at week 48 and 37.7％ at week 96,compared to 33.3％ and 44.8％ in the CHB group.There was no significant difference in these two groups (x2 =1.169,1.424,both P＞0.05).Rate of HBeAg seroconversion in the NAFLD group was significantly lower than that in the CHB group at week 96 (17.9％ vs29.5％,x2=4.972,P＜0.05).Rates of ALT normalization were also found to be significantly lower in the NAFLD group than those in the CHB group at week 48 (69.8％ vs 80.5％,P＜0.05) and at week 96 (82.1％ vs 91.0％,P＜0.05).Multivariate regression analysis demonstrated that serum levels of triacylglycerol (OR=3.725,P=0.000),uric acid (OR=1.005,P=0.040) and fasting blood-glucose (OR=3.067,P=0.000) were independent risk factors associated with NAFLD.NAFLD was found to be a risk factor of virologic response at each time point throughout the treatment.(week 12,OR=2.749,P=0.001; week 24,OR=1.981,P=0.030; week48,OR=2.348,P=0.021; week 96,OR=8.053,P=0.001).Conclusions During antiviral therapy,NAFLD may affect biochemical and serological responses in CHB patients treated with nucleos(t) ide analogues,and may affect early virologic response in these patients.

Objective Blood pressure variability (BPV) is an independent risk factor for the death of patients with maintenance hemodialysis (MHD).There is no study on the influencing factors of BPV at home in HD patients in China.The article aimed to investigate MHD patients'BPV at home and related influencing factors in order to provide theoretical basis for reducing home BPV (HBPV) clinically. Methods We chose 103 patients who were treated with MHD in the Renal Medicine Room of Nephrology Department in three upper first -class hospitals including Jiangsu Provincial People 's Hospital, the First Affiliated Hospital of Suzhou University and the Affiliated Hospi -tal of Jiangsu University from March 2017 to October 2017.We col-lected their 7 days'blood pressure monitoring at home and blood pressure before dialysis, average value and standard deviation in sys -tolic blood pressure monitoring at home, and took the coefficient of variation of systolic blood pressure as the expression of HBPV .The patients were divided into high BPV group (BPV≥5.8％) and low BPV group (BPV<5.8％) according to the average BPV.At the same time, we recorded indexes such as sex , age, dialysis age, primary disease, BMI, inter-dialytic weight gain (IDWG), left ven-tricular mass index(LVMI) and analyzed relative influencing factors by multiple linear regression . Results The age, IDWG and LV-MI were positive independent influencing factors of HBPV (R 2 =0.467,F=10.945,P<0.001).According to standardized regression co-efficient, the contribution of each variable to HBPV was as follows : PIBWG >Age>LVMI. Conclusion In clinical nursing, we should actively control the IDWG of patients , encourage patients to monitor their blood pressure at home , and increase their awareness of the importance of home BPV.Meanwhile, HBPV is an important index for predicting left ventricular hypertrophy and can be used as an objective tool to improve patients 'self-management ability.

AIM To observe the clinical effects of Gushen Antai Pills (Polygoni multiflori Radix,Rehmanniae Radix,Cistanches Herba,etc.) combined with low-molecular-weight heparin (LMWH) in treating women with recurrent spontaneous abortion (RSA) due to coagulation disorder (prethrombotic state,PTS).METHODS One hundred and twenty-six cases of RSA patients in PTS were randomly assigned to observation group and control group for the treatment of either LMWH combined with Gushen Antai Pills or LMWH.The trial went on during the first twenty weeks of pregnancy.Investigations on total treatment efficiency,coagulation-fibrinolysis indicator levels before and after treatment were conducted,and results of follow-ups were compared as well.RESULTS The total effective rate of 73.0％ in the control group and 87.3％ in the observation group displayed a significant difference (P ＜ 0.05).Although the levels of fibrinogen (FIB),D-dimer (D-D) and plasminogen activator inhibitorl (PAIl) were significantly improved (P ＜ 0.05) after the treatment,the observation group was found to be significantly better in performance than the control group (P ＜ 0.05).The live birth rate of 69.8％ in the control group and 85.7％ in the observation group also showed to be a significant difference (P ＜ 0.05).CONCLUSION Gushen Antai Pills combined with LMWH can improve the thrombotic state and bring forth a higher live birth rate in patients with RSA.