In traditional Chinese medicine,medical practice is combined with qi,which forms special qi-theory.Body fluid which includes blood and Jin-ye is the fluid structure of human being.Nutrient qi and defensive qi depend on these two kinds of body fluid.Defensive qi not only maintains physiological functions of Jin-ye,but also participates pathomechanism such as Jin-ye consumption,retention,etc.The identity of defensive qi and Jin-ye plays an important role in theory and clinic.

To study the effect of physiological conditions on spidroins, we analyzed NTR1SR2CT module secondary structure, aggregation and silk-formation influenced by different salts (in different concentration intervals). According to the full-length Araneus ventricosus MiSp sequence, NTR1SR2CT module was constructed and expressed in Escherichia coli BL21 (DE3), and the recombinant proteins were purified by denaturation method mediated by 8 mol/L urea. Random coil and helix are the main secondary structures of NTR1SR2CT and could be induced into beta-sheet by drying natively and lyophilization, where methanol can be used as a promoter. Furthermore, potassium and phosphate cations can cause significant NTR1SR2CT protein aggregation and silk-formation. The results could be a basis for the determination of silk-formation mechanism, and also useful for industrialized generation of high performance spider silk-like fibers.
Animals ; Fibroins ; chemistry ; Protein Structure, Secondary ; Salts ; chemistry ; Spiders

OBJECTIVETo observe and evaluate the clinical performance of zirconia crowns made by CEREC inLab computer aided design/computer aided manufacturing(CAD/CAM) for posterior teeth.

METHODSA total of 242 patients were implanted with zirconia crowns fabricated by CEREC inLab CAD/CAM. The crowns were evaluated by Modified US Public Health Service criteria at baseline, 6, 12, 24, and 36 months. The chi-square test was used to analyze the survival rate.

RESULTSAll but five crowns were evaluated by an independent evaluator at baseline, 6, 12, 24, and 36 months. The survival rate declined with time. The A score percentage was above 85% at 36 months.

CONCLUSIONThe single zirconia crowns fabricated by CEREC inLab CAD/CAM demonstrate satisfactory clinical performance during a short period.

Objective To explore the preventive and therapeutic role of silencing type Ⅰ rat platelet-binding protein motifs depolymerization protein-like metalloproteinase 2(ADAMTS2)by siRNA on experimental liver fibrosis in vitro.By studying the mechanism of siRNA silencing of ADAMTS2,we also aim to evaluate the feasibility of ADAMTS2 as a target for anti-liver fibrosis therapy.Methods Three pairs of siRNAs targeting ADAMTS2 mRNA 2237,2597 and 690 targets were designed and synthesized by utilizing RNA design software.The most effective siRNA was chosen to transfect HSC-T6 cell line to test the tendency of hepatic stellate cell(HSC)activation and ex pression of ADAMTS2,COL1α1,COL(I),α-SMA,TGF-β1,MMP-2 and TIMP-3.These were quantified using real time-PCR,Western blotting,and MTT assays.Results Of the same dosage and time of injection,siRNA 2237 inhibited ADAMTS2 gene expression significantly more than other siRNAs.siRNA-ADAMTS2 2237 markedly inhibited ADAMTS2 gene and protein expression of HSCT6 with more than 80％ efficiency.Conversely,siRNA-ADAMTS2 2237 markedly reduced the gene and protein expressions of COL(I),α-SMA and TGF-β1 on HSC-T6 and inhibited the proliferation of HSC.Conclusions siRNA-ADAMTS2 2237 could effectively knockdown the gene and protein expression of ADAMTS2 in HSC-T6 cell lines.Silencing ADAMTS2 by siRNA significantly inhibited the activation,proliferation of HSC and the gene and protein expressions of COL(I),α SMA,and TGF-β1,and it may have a potential anti-fibrotic effect.ADAMTS2 might be an efficient target for anti-fibrotic therapy.

Objective To explore the effects of inhibiting DDR2 expression by siRNA on hepatic stellate cells and evaluate the role of DDR2 gene in hepatic fibrogenesis. Methods (1) Three pairs of chemically synthesized siRNAs targeting DDR2 were respectively transfected into HSC-T6 cells for evaluation of silence efficacy, and the most effective siRNA was used. (2) HSC-T6 cells were divided into three groups, group A served as normal controls, group B served as negative control and group C was RNA interference DDR2 (siRNA-DDR2) expression of HSC. The most effective RNA interference sequences targeting DDR2 gene was chosen to transfect HSC-T6 cells by plasmid transfection. The tendency of DDR2, α-smooth muscle actin(α-SMA) and collagen-Ⅰ mRNA expression were estimated using RT-PCR, and the protein expression of DDR2 was evaluated by Western blot. Meanwhile, MTT assay was employed to analyze the proliferation of HSC. Results (1) DDR2 siRNA, which began at nt 868, inhibited DDR2 gone expression stronger than the other two siRNAs. (2) After transfection of siRNA-DDR2, the mRNA expression of DDR2 (P<0.01) and α-SMA (P<0.01) significantly decreased compared with the normal group, and the protein expression of DDR2 also significantly decreased (P<0.01). In addition, the proliferation of HSC was also markedly suppressed as compared with the normal group (P<0.01). However, compared with the negative control group, none of them was markedly suppressed. Conclusion SiRNA targeting DDR2 significantly suppresses the activation, proliferation of HSC, and thus attenuates hepatic fibrogonesis in vitro.

Objective To investigate the relation between the expression of Caspase recruitment domain-containing membrane-associated guanylate kinase protein 1 (CARMA1),clinicopathological features of gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma,diffuse large B cell lymphoma (DLBCL),and Helicobacter pylori (H.pylori) infection.Methods From January 1999 to March 2011,34 patients with DLBCL and 20 patients with MALT lymphoma were selected,and at same period 21 cases with reactive hyperplasia of gastrointestinal lymphoid tissue were enrolled in as control.The expression of CARMA1,Ki-67 and cytotoxin-associated gene A (CagA) at protein level were examined by immunohistochemistry.The relative expression quantity of CARMA1 mRNA was detected by real-time polymerase chain reaction (PCR).The condition of H.pylori infection in 25 gastric lymphoma and 10 controls was detected by methylene boric acid and blue staining or semi-nested PCR.Chi-square test was used for counting data analysis,t test for measurement data.Multivariate COX regression analysis was implemented for survival analysis.Survival curve was drawn by Kaplan-Meier method and analyzed by Log-rank test.Results The relative expression quantity of CARMA1 mRNA of 28 patients with DLBLC (3.073±1.846) was higher than that of 14 patients with MALT lymphoma,and the difference was statistically significant (F 0.975,P＜ 0.05).The positive rate of CARMA1 expression at protein level of gastrointestinal lymphoma group (75.9 ％,41/54) was higher than that of control group (47.6％,10/21),and the difference was statistically significant (x2 =5.568,P＜0.05),and the positive rate of CARMA1 expression of MALT lymphoma group (11/20) was lower than that of DLBCL group (88.2％,30/34),and the difference was statistically significant (x2 =5.900,P＜0.05).Among 42 patients with gastrointestinal lymphoma who received surgery,the relative expression quantity of CARMA1 mRNA in cases with high proliferation (2.885±1.837) was higher than that in cases with low proliferation.The expression of CARMA1 mRNA in the cases at advanced stage of the disease (4.416± 1.010) was higher than that in cases at early stage,and the difference was statistically significant (F=3.317 and 2.972,both P＜0.05).Among 54 patients with gastrointestinal lymphoma,the positive rate of CARMA1 expression at protein level of patients with high proliferation (88.6％,31/35) was higher that that of patients with low proliferation (10/19),and the difference was statistically significant (x22 ＝ 6.847,P＜0.05).There were no significant differences in relative expression quantity of CARMA1 mRNA and the positive rate of CARMA1 expression at protein level between 11 gastric lymphoma patients without H.pylori infection and 14 gastric lymphoma patients with H.pylori infection (both P＞0.05).The positive rate of CARMA1 expression at protein level of CagA positive and CagA negative H.pylori infected gastric lymphoma patients was 11/11 and 2/3.The expression of CARMA1 at protein level was correlated with the prognosis of gastrointestinal lymphoma (RR＝4.160,P＜0.05).In the 29 cases of patients with gastrointestinal MALT lymphoma and 18 cases of patients with DLBCL who were followed up,the survival situation of gastrointestinal lymphoma patients with CARMA1 positive expression rate over 50％ was worse than that of patients with the rate less than 50％,and the difference was statistically significant (x2=5.383 and 4.028,both P＜0.05).Conclusions CARMA1 might be involved in the pathogenesis and progression of MALT and DLBCL; and it might be a related factor of poor prognosis.There was no correlation between the expression of CARMA1 and H.pylori infection in these two lymphomas.

AIM: To establish the quality standard for Tongfeng Huadu Tincture (Radix Gentianae Macrophyllae, Radix Angelicae Pubescentis, Rhizoma Chuanxiong, Herba Asari, etc.). METHODS: The Radix Gentianae Macrophyllae, Radix Angelicae Pubescentis, Rhizoma Chuanxiong, Herba Asari of Tongfeng Huadu Tincture were identified by TLC. Strychnine of Tincture was determined by HPLC. RESULTS: The average recovery was 97.07% and RSD and 2.1% (n=5). CONCLUSION: The method is reliable, accurate and specific. It can be used for quality control of Tongfeng Huadu Tincture.

Objective To determine the diagnostic value and clinical significance of sonographically detected fetal dysplastic kidney with normal amniotic fluid volume. Methods At the 2nd or 3rd trimester of gestation,the fetuses with unilateral or bilateral renal anomalies (ahnormal size,echo,shape or cyst of the kidney) and normal amniotic fluid volume received systemic ultrasound examination,autopsy or follow-up until after birth. The fetus with only dilated renal pelvis was not included. Results Eleven fetuses of dysplastic renal anomalies with normal amniotic fluid volume were identified by prenatal ultrasound. Among the five fetuses affected by unilateral multicystic kidney dysplasia (MCKD),the renal anomaly was isolated in four fetuses,and the other one was complicated with absence of the ipsilateral hand. One of the two fetuses of unilateral renal agenesis had no other associated anomaly and the other one was complicated with hydrocephalus,spina bifida,ipsilateral absent radius and single umbilical artery,correspongding to the VACTERL syndrome. Two fetuses of pelvic kidney and horseshoe kidney respectively was proved by postnatal ultrasound. One fetus was diagnosed as autosomal dominant polycystic kidney disease(ADPKD)on the basis of multiple renal cysts and a positive family history,the fetus also had cardiac rhabdomyoma. One fetus of bilateral normal sized hyperechoic kidneys was proved to be renal dysplasia by autopsy. Conclusions Unilateral MCKD is the most common type of fetal renal dysplasia which can be detected by prenatal ultrasound with normal amniotic fluid volume. Based on the sonographic characteristics and the family history,most of the dysplastic renal anomalies can be diagnosed prenatally and the prognosis can be predicted.

Purpose To investigate the clinicopathological characteristics and the survival outcomes of invasive lobular carcinoma. Methods A retrospective analysis of 98 patients with invasive lobular carcinoma and 530 invasive carcinoma of no special type was performed in order to observe the histological features and the clinical outcomes of invasive lobular carcinoma. Results Median follow-up was 68. 5 months for invasive lobular carcinoma and 67 months for invasive carcinoma of no special type. Invasive lobular carcinoma presented with a larger tumor size, more histopathological grade 2 tumors, increased rate of hormonal receptor positivity, human epider-mal growth factor 2 (HER-2) negativity, and had a lower proliferative index as compared to invasive carcinoma of no special type, more frequently presented with the luminal A subtype (P<0. 001). The classical invasive lobular carcinoma presented with a smaller tumor size, to have a lower histological grade and proliferative index compared to the non-classic type, and more frequently presented with the luminal A subtype, whereas the non-classic invasive lobular carcinoma patients more frequently presented with the luminal B, HER-2 overexpression, or triple negative subtype (P=0. 035). A statistically significant difference in the outcome was observed at un-ivariate analysis for patients with non-classic for disease-free survival (P=0. 043) and for overall survival (P=0. 048), as compared with patients with classical invasive lobular carcinoma. The disease-free survival difference between the invasive lobular carcinoma and the invasive carcinoma of no special type was not significant (P=0. 537), and the overall survival rates were not statistically different between the two groups (P=0. 397). A statistically significant difference of overall survival was observed at multivariate analysis for patients with HER-2 positive and triple negative subtypes versus patients with luminal A invasive lobular carcinoma (P=0. 015, P=0. 016) . Conclusions The outcome of invasive lobular carcinoma is significantly correlated with histological and immunohistochemi-cally defined molecular subtypes. New tailored strategies should be explored in these subgroups of patients with poor outcome.