ITS2 sequence was used as a barcode to identify Viticis Fructus and its adulterants. The study collected 46 samples include Viticis Fructus and its adulterants. The sequences were obtained by extracting DNA, amplification, sequencing bi-direstionlly, and then assembling with CodonCode Aligner. To identify Viticis Fructus and its adulterants, the study used the methods of computing the genetic distances by Kimura 2-Parameter (K2P) model and constructing the Neighbor-joining (NJ) phylogenetic trees using MEGA5.0. The results showed that the maximum inter-specific K2P distance of Viticis Fructus was less than the minimum inter-specific K2P distance of Viticis Fructus and its adulterants and the NJ tree indicated that the Viticis Fructus and its adulterants could be distinguished clearly. Therefore, using ITS2 barcode can distinguished Viticis Fructus and its adulterants accurately.

ObjectiveTo analyze the change in the incidence of papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC) in the past 27 years and to probe the relationship between iodine and these carcinomas. MethodsStatistical analysis was performed on data of thyroid diseases, thyroid malignant tumors, PTC , FTC and their complications in Department of Pathology from 1978 to 2004 in General Hospital of Tianjin Medical University with Run test、Cox-Stuart test, ANOVA, t or t′ test and Chi-square test. ResultsIn 27 years there were 231837 cases of biopsies (average 8587cases/year). The biopsies were performed in the thyroid diseases as a whole 6276 cases (2.70%),thyroidmalignanttumors644cases(0.28%), PTCs 465 cases (0.20%), and FTCs 80 cases (0.03%) in 27 years. Biopsies in thyroid diseases as a whole showed a descending tendency since 1982 but an ascending tendency after 1995. Biopsy in malignant thyroid tumor did not show a significant change, and biopsy in PTC showed an ascending tendency and that in FTC showed a descending tendency after 1995. The average age, both in male and female patients, at the time of diagnosis of PTC and FTC showed no significant change during 27 years, but the mean age of male at the time of diagnosis was older than that of female. The ratios of male to female in PTC and FTC were 1:3.31、1:2.81 respectively,and showed no significant change either. There were more biopsies with lymph node metastasis in PTC (33.76%) than those in FTC (10.00%) and more biopsies with blood vessel involved or nodular goiter in FTC (11.25% and 13.75%) than those in PTC (0.65% and 4.52%). Mean age of patients with PTC and nodular goiter (44.3 years old) was older than those without goiter (38.1 years old). The ratios of male to female with PTC and FTC with lymph node metastasis were higher than those without lymph node metastasis. ConclusionThe incidence of thyroid malignant tumors in 27 years has no significant change. The contrary tendency of PTC and FTC suggests that the decreasing malignant degree of carcinoma. It suggests that this area is not in high iodine condition and the change have no relationship with the iodine intake.

Objective To investigate the effects of MMI-166 on apoptosis and apoptosis-related protein expression of human pancreatic cancer SW1990 cells and its transplanted tumor,and explore possible mechanism.Methods The human pancreatic cancer xenograft model was constructed by using human pancreatic cancer SW1990 cells.Tumor-bearing nude mice were randomly divided into control and MMI-166 groups,and they were treated with normal saline or MMI-166 (200 mg · kg-1 · d-1) for 28 days.Apoptosis index (AI),p53,c-Myc,Bax,Bcl-2,Survivin,Caspase-1,Fas proteins were detected by deoxynucl-eotidyl transferase-mediated nick end labeling (TUNEL method) and Western blot.MMI-166 of different concentrations (0,50,100 μg/ml) were used to treat human pancreatic cancer SW1990 cell for 24 h.The c-Myc,Survivin proteins expressions were measured by Western blotting.Results Apoptosis index in MMI-166 group was 81.1 ±7.9,which was significantly higher than that in control group (21.3 ±2.2,P =0.000) ; the expressions of c-Myc,Survivin were 7715 ± 2229,4594 ± 1240,which were significantly higher than those in control group (16870 ± 2446,15208 ± 1903,P =0.000) ; the expressions of p53,Bax,Bcl-2,Caspase-1,Fas were not significantly different from those in control group.After 50,100 μg/ml MMI-166 treatment,the expression of c-Myc was significantly down-regulated (0.098 ± 0.003,0.073 ± 0.008 vs.0.169 ± 0.007,F =189.361,P ＜ 0.05) ; and the expression of Survivin was not significantly changed.Conclusions MMI-166 may induce cell apoptosis of SW1990 by down-regulating the expression of c-Myc.

Objective To observe the effects of herbal compound in reinforcing the kidney for activating blood and arousing consciousness(HCRAA) on the ABR threshold and the expression of neurotrophic factor 3(NT-3) of the cochlea of gentamicin (GM)-induced ototoxic mice.Methods 40 mice were randomly divided into normal group,model group,and high,middle and low concentration HCRAA groups.Normal group received no treatment.The model group and the HCRAA groups were intraperitoneally injected with GM 100mg/kg per day for consecutive 15 days.At the same time,the model group and the HCRAA groups respectively receiveded normal saline and high,middle and low concentration Chinese medical formula decoction at the same dosage by garage for consecutive 20 days.After the experiment,the mice were tested ABR.The expression of NT-3 of the cochlea in mice was detected by western blot.Results HCRAA at high and middle concentration reduced the GM elevated ABR threshold(P＜0.01),and increased the expression of NT-3 in the cochlea(P＜0.01).Conclusion HCRAA may effectively reduce the elevated ABR threshold induced by gentamycin by protecting GM damaged cochlear hair cell and neurons and increasing the expression of NT-3 in the cochlea.

Non-small-cell lung carcinoma (NSCLC) is one of the most frequently diagnosed malignancies worldwide.Previous studies have shown that microRNA-449b (miR-449b) functions as a tumor suppressor in many cancers.However,the role of miR-449b in NSCLC is still unknown.In the present study,miR-449b was significantly downregulated in NSCLC samples and cell lines.Bioinformatics analysis revealed that 3'-UTR region of leucine rich repeat containing G protein-coupled receptor 4 (LGR4) mRNA had putative complementary sequences to miR-449b,which was further confirmed by the luciferase assay.Western blotting showed that restoration of miR-449b in NSCLC cells decreased the expression of LGR4.Interestingly,over-expression of miR-449b inhibited growth and invasion of NSCLC cells in vitro.Furthermore,ectopic expression of LGR4 reversed miR-449b-suppressed proliferation and invasion of NSCLC cells.Therefore,the data of the present study demonstrate that miR-449b inhibits tumor cell growth and invasion by targeting LGR4 in NSCLC.

Normal function of growth hormone-insulin-like growth factor Ⅰaxis is essential for linear growth after birth. A case of continuous growth with undetectable growth hormone level even under insulinhypoglycemia stimulation was reported. The growth hormone deficiency was due to pituitary stalk interruption combined with deficiency of multiple pituitary hormones. Taken together with reviewed literature, this so-called nongrowth hormone-dependent linear growth was preconditioned by other hormones, especially gonadotropin deficiency,and the unclosed epiphysis.

Objective To generate recombinant Pichia pastoris for high-copy expression of human tissue factor pathway inhibitor (TFPI). Methods The cDNA encoding human TFPI was inserted into the expression vector pPIC9K and the constructed expression vector rhTFPI-pPIC9K was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant plasmids were subsequently transformed into Pichia pastoris GS115 cells, and the transformants were confirmed by PCR amplification of the genomic DNA.The recombinant Pichia pastoris with high copies of TFPI cDNA was screened by G418 selection. Western blot and TFPI ELISA Kit were employed to analyzing. The temperature, time and concentration of methanol for the induction of recombinant protein were optimized. Results PCR analysis and DNA sequencing confirmed the successful construction of the expression vector rhTFPI-pPIC9K. Real time quantitative PCR and Western blot analysis demonstrated the positive correlation between TFPI expression level and the copy number of TFPI cDNA in Pichia pastoris cells. Optimization of the induction condition significantly elevated the expression level and activity of TFPI (9.95±0.78 mg/L and 3.91±1.37 U/mL). Conclusion The Pichia pastoris strain with high copy of TFPI expression was successfully constructed, which lays a solid foundation for the further investigation on the function of TFPI and its application in the prevention and therapy of diseases.

Objective To investigate the therapeutic effect of Jianpi Yiqi, Bushen Fenjing combined with oxaliplatin+CF/5-FU on colorectal cancer.MethodsEighty patients with colorectal cancer who were treated in our hospital from January 2014 to December 2015 were randomly divided into observation group(40cases) and control group (40cases).The control group was given oxaliplatin+CF/5-FU treatment, the observation group in the control group based on the application of Spleen Qi, Bushen fill essence treatment, observed before and after treatment of two groups of serum tumor markers, clinical symptoms and living conditions, changes, record adverse reactions.ResultsThe observation effect of 82.50% was significantly higher than that of the control group (62.50%).The QOL score (37.8±5.6) was significantly higher than that of the control group (33.5±3.1) and the adverse reaction rate was 30.00% (P<0.05).The difference between the two groups was significant (P<0.05).ConclusionThe effect of invigorating spleen qi and tonifying siren solution combined with oxaliplatin+CF/5-FU on colorectal cancer can improve the clinical symptoms and improve the quality of life of patients.

Objective To evaluate the Carryover between the chemistry and immunoassay in Beckman Coulter Laboratory Au-tomation System and decide to whether sharing samples for testing between chemistry and immunoassay systems or not. Methods According to a certain order,high concentration samples and low concentration samples of HCG with different sample volume (500 μl,2 000 μl)were tested on Beckman AU5421 automatic biochemical analyzer.The HCG of low concen-tration samples were then tested to evaluate the carryover between the chemistry and immunoassay and explored the correc-tive procedure to deal with the carryover by increasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.Results Under different sample volume,the carryover in a single module and as a whole of the beckman AU5421 automatic biochemistry analyzer were 5.44,15.47,23.51 and 45.96 ppm respectively (t=14.553,P <0.001;t=5.527,P =0.005;t=3.985,P =0.016;t=20.457,P <0.001).By increasing special cleaning process the carryover of 0.22 ppm was detected in 500 μl sample volume of the beckman AU5421 automatic biochemistry analyzer as a whole.Conclusion The car-ryover between the chemistry and immunoassay in Beckman Coulter Laboratory Automation System could been sovled by in-creasing special cleaning process of beckman AU5421 automatic biochemistry analyzer.

ObjectiveTo investigate the correlation between electro-acupunctured point Houxi(SI3)and activated brain function areas, contrast the images produced with electroacupuncture at left and right points Houxi and make a comparison with electroacupuncture at point Hegu(LI4)in patients with peripheral facial paralysis.MethodPatients with peripheral facial paralysis, six on the left side and six on the right side, were enrolled as subjects. A scan of the whole brain was taken using fMRI during electroacupuncture stimulation. The images were processed using SPM software. An analysis using at-test (P<0.01) showed differences in brain functional images produced with electroacupuncture at different points.ResultElectroacupuncture at left point Houxi increased the signals of brain regions: rightcaudate nucleus, right cingulate gyrus, right parahippocampal gyrus, right superior temporal gyrus, the brainstem and the cerebellar vermis. Electroacupuncture at right point Houxi increased the signals of brain regions: right medial frontal gyrus, left middle frontal gyrus, left anterior cingulate gyrus, rightcingulate gyrus and right superior temporal gyrus.ConclusionThere is a difference in the image between electroacupuncture at point Houxi and at point Hegu or Dicang(ST4). There is also a larger difference in the image between bilateral points Houxi. The brain regions with high-frequency or low-frequency signals are not consistent. The results further prove the scientificalness of “Point Hegu is indicated for diseasesin the face and mouth” and also show that the cognominal acupoints on the two sides ofthe human body may have some differences, e.g. the conduction pathways are not completely the same and the therapeutic effects are not completely consistent.