Objective To study the influence on STR typing for low copy number(LCN)DNA using different methods of amplifcation and detection.Methods Control DNA 9947A was diluted and then amplifled with Identifiler~(TM) or DNATyper15~(TM).The heat cycles were set to 28 or 28 add 6.Each template wag amplified three times in parallel,and then the amplified products or the product mixture of three amplifictions were deteced with 310 or 3130 Genetic Analyzer.Results The success rate of STR typing with the method 28+6 cycles was higher than that of 28 cycles.There are no correlation between the allele imbalance or allele dropout and STR locus.With the reduction in the amount of template DNA,allele imbalance and dropout gradually increased,and the allele dropout was more common than allele imbalance when the amount of template DNA Wag very small.When the product mixture of three amplifictions were deteced.the occurrence of allele imbalance and dropout reduce obviously.Conclusion The success rate of STR typing of LCN DNA can be obviously increased by detecting the product mixture of three amplifictions in parallel combined with the 28+6 heat cycle condition.

In order to study the structure-function relationship in the protein which is incorporated with p-nitro-L-phenylalanine,the method of MD(Molecular Dynamics) simulation was established and successfully used in the analysis of protein which contains p-nitro-L-phenylalanine.The force field of CHARMM can only stimulate protein with natural amino acid in NAMD.Compared with phenylalanine,p-nitro-L-phenylalanine just has one more group of nitro.If the parameter of group of nitro was defined,the protein containing p-nitro-L-phenylalanine can be simulated.CGenFF-paramchem was used to calculate the energy and topological structure of p-nitro-L-phenylalanine' s new bonds (r),angles (θ),dihendrals (φ) and improper angle (ψ).And then the new defined parameter and topology information was input into the related parameter files and topology files in CHARMM.On the basis of correct parameter,NAMD can successfully simulate the modified BAFF(B lymphocyte stimulator) which contains p-nitro-L-phenylalanine.The changes in structure indicated that there might be new B cell epitopes.The temperature distribution of each frame in the process of dynamics stimulation was in accord with normal distribution,which proved the defined force field parameters was feasible.The RMSD of whole protein solution systemis 2.5.Calculate each resides' RMSF in BAFF,the RMSF of p-nitro-L-phenylalanine's residue is 3.7,which is obviously higher than that of the other residues in β-pleated sheet,and close to the loop rings,indicate that there might be variation in the area of p-nitro-L-phenylalanine residue and might produce new comformational epitopes.The results of MD stimulation will guide the immunogenicity experiments of p-nitro-L-phenylalanine modified proteins.

Objective To observe the effect of Ginkgo on IL-4,IL-5 in serum of asthmatic mice. Methods 40 mouse were randomly divided into three groups(control group, sensitization group, and therapeutic group). The mouse asthmatic models were established by sensitizing with aerosolized ovalbumin(OVA). IL-4,IL-5 in serum in 16 sensitization mouse and 8 control mouse were measured by ELISA, respectively. We injected 0.15ml Ginkgo solution in the abdominal cavity of 16 therapeutic mouse before the episode of asthma. After that, they were challenged with aerosolized OVA and IL-4,IL-5 in serum were measured by ELISA. Results The levels of IL-4,IL-5 in serum in control group were (12.965?2.450)pg/ml, (13.398?4.460)pg/ml, respectively, (16.889?4.835)pg/ml, (19.471?7.207)pg/ml in sensitization, (13.695?4.068)pg/ml, (14.774?5.749)pg/ml in therapeutic group. In the sensitization group, the levels of IL-4,IL-5 in serum were higher than those of the control group( P

OBJECTIVETo observe and evaluate the clinical effect of acupuncture at "Siguan" combined with the Gallbladder Meridian acupoints on migraine.

METHODSWith randomized controlled trial design, 87 patients were randomly divided into an observation group (42 cases) and a control group (45 cases). The observation group was treated with acupuncture at "Siguan" including Hegu (LI 4) and Taichong (LR 3), combined with the Gallbladder Meridian acupoints including Fengchi (GB 20), Shuaigu (GB 8), Qiuxu (GB 40) and Zulinqi (GB 41), while the control group was treated with acupuncture at the Gallbladder Meridian acupoints, including Fengchi (GB 20), Shuaigu (GB 8), Qiuxu (GB 40), Zulinqi (GB 41), Yanglingquan (GB 34) and Hanyan (GB 4). The treatment was given every other day in two groups, 3 times per week for totally 6 weeks. The migraine intensity, migraine frequency and lasting time of each migraine were observed before and after treatment, and 3-month follow-up was performed after treatment. The clinical efficacy in two groups was evaluated.

RESULTSCompared with those before treatment, the migraine intensity, frequency and lasting time were all reduced significantly after treatment in the two groups (all P<0.01), which were more obvious in the observation group (all P<0.05). At the end of the treatment, the total effective rate was 95.2% (40/42) in the observation group and 80.0% (36/45) in the control group, which had statistical difference (P<0.05). After 3-month follow-up, the total effective rate was 88.1% (37/42) in the observation group and 75.6% (34/45) in the control group, which had no statistical difference (P>0.05).

CONCLUSIONThe "Siguan" combined with the Gallbladder Meridian acupoints could significantly relieve the migraine intensity, frequency and lasting time, and its improvement of symptoms and short-term efficacy are superior to those of the Gallbladder Meridian acupoints alone.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Female ; Gallbladder ; physiopathology ; Humans ; Male ; Meridians ; Middle Aged ; Migraine Disorders ; physiopathology ; therapy ; Young Adult

Objective To assess the regional left ventricular (LV) rotation motion in the patients with rheumatic mitral stenosis (MS) with velocity vector imaging (VVI).Methods This study included 38 patients with isolated MS (mild,moderate and severe) and 55 healthy control subjects.Short-axis parasternal views at the basal and apical level of LV were taken.Peak value of rotation angle and rotation velocity,as well as time to peak value were measured by VVI workplace offline respectively.Results ①In the healthy control subjects LV rotates clockwisely at the base and counterclockwisely at the apex,the same as the patients with MS.② In th patients with MS,rotaion angle and rotation velocity were significantly reduced at the apical level(P ＜0.001),rotation velocity were significantly reduced at some segments of basal level (anterior,lateral,septal) (P ＜ 0.05).③Compared with the healthy control,the difference of time to peak roation angle between base and apex was significantly longer in patients with MS (P ＜0.05).④ There were no significant differences among mild,moderate and severe MS (P ＞0.05).Conclusions The patients with MS may suffer from systolic dysfunctin even in the early stage identified by the alteration of LV rotation motion,which is independent of the hemodynamic severity of MS.

Objective To investigate the role of mismatch repair gene hMLH1 in pancreatic carcinoma and its clinicopathological significance.Methods hMLH1 was extracted from 60 cases of pancreatic carcinoma tissues and 60 cases of normol pancreatic tissues.hMLH1 expression in pancreatic carcinoma and normal tissues was detected by SP immunohistochemical staining.Results The strong,weak and loss expression of hMLH1 in pancreatic carcinoma tissues and normal pancreatic tissues was 0 vs 83.33％ (50/60),31.7％ (19/60)vs 16.67％ (10/60),and 68.3％ (41/60) vs 0 respectively.The protein expression of hMLH1 was not related to patient's age,tumor location,or pathological types (P ＞ 0.05),but it was related to lymph node metastasis (x2 =8.579,P =0.004),clinical stage (x2 =9.586,P =0.002) and pathological differentiation (x2 =20.372,P =0.001).Conclusion The loss expression of hMLH1 has a correlation with pancreatic carcinogenesis,differentiation degree,and disease progression.

OBJECTIVE: To investigate the therapeutic efficacy of treatment of Bacillus licheniformis (BL) on ulcerative colitis(UC) in BALB/c mice. METHODS: The therapeutic effect of different treatment was evaluated in seven acute UC mice groups and one control group, including the normal group, the model group, the solvent group, the sulfasalazine (SASP) treated group, the high dose BL treated group, the low dose BL treated group, the high dose BL with SASP treated group and the low dose BL with SASP treated group. The disease activity index (DAI) score of mice were recorded every other day since the experiment started. On the 15th day, all the mice were killed, then gross injury score in colonic mucosa and histopathological changes were measured. The expression levels of NF-κB and Bcl-2 in mouse colonic mucosa was also determined by immunohistochemistry method. RESULTS: The DAI score, the colonic gross injury score, the histopathological changes and the expression level of NF-κB and Bcl-2 of mice colonic mucosa in drug treated groups were decreased significantly when comparing with the drug untreated groups. Moreover, these results were even better in two drugs combined treated groups than in single drug treated groups (P<0.05). CONCLUSION: BL has the efficacy of remission DSS-induced acute ulcerative colitis in mice, and there is more effective when it is administered with SASP. The results indicat that the therapeutic effort is via inhibiting the expression of NF-κB and Bcl-2 in colonic mucosa.

Objective To establish the quality control standard for Shuanggen Qingnao Granules.Methods TLC was used to identify Yiyiren SEMENCOICIS,Yujin RADIXCURCUMAE,Banlangen RADIX ISATIDIS. HPLC was used to determine the content of Puerarin in Shuanggen Qingnao Granules.Results The TLC spots developed were fairly clear,and the bland test showed no interference.Puerarin showed a good linear relationship in the range of 5.184～40.50?g/mL,the average recovery of Puerarin was 99.93%,and RSD=1.81%.Conclusion The method is accurate and quick,and can be used for the quality control of Shuanggen Qingnao Granules.

Objective To investigate the association of human leukocytes antigen(HLA)-DR/DQ with oral lichen planus(OLP) in the area of Yangtze River Delta. Methods HLA-DRB1 and DQB1 genotyping of 44 unrelated OLP patients and 150 normal controls were performed by polymerase chain reaction-sequence specific primers(PCR-SSP) method.The data were compared between the OLP group and normal controls,and between different types of OLP patients. Results The frequency of HLA-DRB1*09 and HLA-DRB1*07 alleles were significantly higher in OLP group than those in normal controls(56.8% vs 31.3% and 27.3% vs 13.3%,P

Objective To investigate the synergistic effects of 9-cis-retinoic acid(9-cis-RA) and 8-cl-cAMP on growth inhibition and apoptosis induction in H460 and H292 cell lines of non-small-cell lung cancer(NSCLC). Methods Experimental groups included 9-cis-RA groups(1,5,10 and 20 ?mol/L),8-cl-cAMP groups(5,10,20 and 50 ?mol/L),9-cis-RA(5 and 10 ?mol/L) combined with 8-cl-cAMP(10 ?mol/L) groups and blank control group.The cell growth inhibition rates were detected by trypan blue staining,and the apoptosis of H460 and H292 cells were observed by Hoechst33258 fluorescence microscope,DNA gel electrophoresis and flow cytometer(FCM). Results 9-cis-RA inhibited the growth of H460 cells in a time-and dose-dependent manner,and induced the apoptosis of H460 cells(P