C2H2 zinc-finger motif presents in 3% of proteins that are encoded in the human genome, and has the abilities to recognize DNA, RNA and protein. With nearly 3 decades of efforts, the mechanisms of zinc-finger mediated biomolecule recognitions have been studied to various extents. Zinc-finger binds into the major groove of DNA double helix, establishes an one-to-one recognition format between DNA bases and certain amino acids in a zinc-finger, and achieves specificity based on DNA sequences. While RNA molecules show a large variety in their structures, zinc-finger recognizes RNA through the collected information of specially displayed bases and special backbone folding. Initial studies have been performed on zinc-finger mediated protein-protein interactions. Existing data indicate multiple recognition modes. The studies on molecular mechanism have supported the development of engineered zinc-fingers, which have been introduced into applications. For its wide existence, large functional diversity and potential in translational applications, zinc-finger deserves a systematic study in every aspect.

Brother of regulator of imprinted sites (BORIS), also known as CCCTC binding factor-like (CTCFL), is a relatively newly discovered cancer-testis antigen. Drug development related to BORIS has been carried out in multiple directions, including small molecules, small RNA molecules, polypeptides, vaccines, and cellular therapies. Due to its unique ability to interfere with the higher-order spatial structure of the genome, BORIS may represent a new class of drug targets. Here we systematically review the molecular biology research results related to BORIS, including the diversity of its gene products, the multipartite interactions mediated by BORIS, the subsequent impact on signaling pathways, and current drug development strategies, in order to gain a better understanding of the molecular mechanisms of BORIS in both upstream and downstream regulation networks and to identify potential research directions for further breakthroughs.

AIM: To evaluate the diagnostic accuracy of macular ganglion cell - inner plexiform layer ( GCIPL ) measurements using high- definition optical coherence tomography ( Cirrus HD - OCT ) ganglion cell analysis algorithm for detecting early and moderate to severe glaucoma.
METHODS:Twenty normal control persons, 26 patients with early glaucoma and 29 patients with moderate to severe glaucoma were enrolled in this study. Macular GCIPL, optic nerve head ( ONH ) parameters and peripapillary retinal nerve fiber layer ( RNFL ) thickness were measured in each subject. Then all measured results of each parameter were calculated using SPSS17. 0. Areas under the receiver operating characteristic curves ( AUC) of each parameter were calculated to compare the diagnostic accuracy for detecting early and moderate to severe glaucoma.
RESULTS: For detecting early glaucoma, AUC of average RNFL and seven clock value of RNFL were the biggest ( 0. 871 and 0. 896 respectively ), the AUC of parameters in GCIPL were also significant, among them,
the average GCIPL showed bigger AUC(0. 847) than the minimum GCIPL (0. 812). For diagnosing moderate to severe glaucoma, the AUC of rim area was 0. 992, which was bigger than that of average RNFL ( 0. 991 ). The minimum GCIPL showed bigger AUC ( 0. 983 ) than the average GCIPL (0. 967). For early glaucoma diagnosis, the sensitivity of average RNFL was the highest (76. 9%), while the average GCIPL has the highest specificity (93. 5%).
CONCLUSION: AS a new diagnostic parameter for detecting glaucoma, GCIPL shows similar diagnostic potential compared with RNFL. For early glaucoma diagnosis, average RNFL is the most important parameter, while screening early glaucoma, average GCIPL should be paid more attention.

Objective To investigate the expression and signification of B-cell lymphoma/leukemia-2(bcl-2),bax and ER proteins in epithelial cells in different depth of adenomyosis.Methods Expression and correlation of bcl-2,bax and ER proteins were detected by immunohistochemistry staining in 36 adenomyosis cases with superficial (＜ 1/2 depth of penetration) and deep (≥ 1/2 depth of penetration)ectopic endometrium and eutopic endometrial tissues matched with 30 cases with benign ovarian tumor,uterine septum,pelvic floor dysfunction without adenomyosis as controls.Results (1) The expression of ER protein in superficial and deep ectopic tissues (5.04 ±0.24,4.91 ±0.16) were found significantly lower than 6.06 ± 0.36 in eutopic tissues (P ＜ 0.01) and higher than 3.70 ± 0.58 in control group (P ＜ 0.05).There was no statistical difference of ER expression in superficial and deep ectopic endometrium (P ＞ 0.05).(2) The level of bcl-2 protein of 5.6 ± 0.4 in superficial and 6.0 ± 0.3 in deep myometrium of ectopic tissues were significantly higher than 3.6 ± 0.4 in eutopic tissues and 1.9 ± 0.4 in control group (P ＜ 0.01).(3) The level of bax protein of 3.50 ± 0.28 in superficial and 4.80 ± 0.29 in deep myometrial ectopic and 4.43 ± 0.37 in eutopic tissues were significantly lower than 6.18 ± 0.65 in control groups (P ＜0.05).The expression of bax in superficial myometrium is significantly lower than deep myometrium in ectopic tissues (P ＜0.01).(4) The positive correlation between the expression of ER and bcl-2 protein at superficial myometrium of ectopic tissues (r =0.720,P ＜ 0.01).And there was no significant correlation with the expression of ER and bax protein at superficial myometrium (r =0.008,P ＞ 0.05).As well as,there was not significant correlation with the expression of bcl-2,bax and ER protein at deep myometrium (r =0.089,r =-0.023,P ＞ 0.05).The expression of bax protein in ectopic endometrium was positive correlation with the depth of adenomyosis penetration (r =0.736,P ＜ 0.01).There was positive correlation between the expression of ER and bcl-2 protein at normal endometrium (r =0.453,P ＜ 0.05).And there was negative correlation between the expression of ER and bax protein at control group (r =-0.514,P =0.05).Conclusion The bax protein expression of ectopic endometrium in deep adenomyosis was higher than superficial adenomyosis.

Objective To make a comprehensive evaluation of nano-composite of poly(L-lactide) and surface grafted hydroxyapatite(PLLA/PLLA-gHA) as a new material.Methods According to the evaluated critera of medical implanted materials biology and animal trial recommended in GB/T 16886 and IS0 10993 criterion,the new material was carried out on acute systemic toxicity test,haemolysis test,muscular implantation test and subcutaneous injection test.The extract liquid of new material was injected into mice by vena caudalis to test common station,toxic reaction of it at different time,the results were used to evaluate the acute systemic toxicity.Fresh anticoagulant cony blood was mixed with extract liquid of new material with density of 100 g?L-1 to measure each absorbance with spectrophotometer and work out the corresponding rate of haemolysis.The red punctuation and hydropsia of rabbits were observed at different time by subcuntaneous injecting extract liquid into the back of rabbits.PLLA/PLLA-gHA composite plates were implanted into the sacrospinal muscle of rabbits.Cony venous blood was extracted to detect indicatrix of hematology at diferrent time.The material and surrounded tissues were taken out from animals at the 14th,30th,60th,90th,180th,360th day to examine anatomic and pathological changes.Results Rabbits with PLLA/PLLA-gHA composite had good general condition.There was no any acute systemic toxicity in vivo.Data of AST and Scr had no significant difference between experimental group and control group.The hemolysis rate of extrac liquid was 1.22%,which was under the standard criteria(5%).No red punctuation and light hydropsia were observed at different time in the subcutaneous injection test.The inflammation cytochange of PLLA/PLLA-gHA composite group was similar with that of control group in early days,which was met with the general regularity of inflammatory outcome.The fibrosis membrane surrounding the PLLA/PLLA-gHA composite became thinner gradually with the elongation of implantation time.The fibrosis membrane grew into the material at the 360th day.The degree of the fibrosis membrane was below class Ⅰ.Conclusion The new absorbable type PLLA/PLLA-gHA composite has excellent biocompatibility and security.

Objective To investigate the effects of ginsenoside Rg3 (GS-Rg3) on the hypertrophic scars of rabbit ears and provide experimental foundation for study on its inhibition on the hypertrophic scars. Methods Hypertrophic scars were proved on 24 white rabbits,of which the whole level of the skin was excised for 2 cm?2 cm,4-6 points for each ear,controlled by itself. GS-Rg3 0.1 mL(concentration 3 g?L-1) was injected into experimental group and the same volume of saline solution into control group,once every three days regionally. The scar tissues were collected 2,4 and 6 weeks after the injection respectively,the thickness of the scar,structure under the microscope,and the expressions of PCNA,Bcl-2 and Bax were observed. Results In control group,three weeks after the epithelization of the wound,the thickness of the hypertrophic tissue was 3-4 times of ventro ear skin. Under microscope,the dermis was hyperplasia and got thicker,consisted with amount of fibroblast cells,collagen and vessels,the collagen was untidy,nodule or vortex,and the cartilage could be observed in some region.In experimental group, six weeks after the injection,the skin got thinner,the collagen became neath and the quantity of the vessels decreased. In the hypertrophic scars,there was high expression of PCNA,the percent of positive cells was higher (39.55%?6.07%) compared normal tissue (11.18%?1.71%).In GS-Rg3 group,the expression of Bcl-2 was gradually decreased two weeks after injection and obviously decreased six weeks later,there was significant difference compared with before injection (P

This paper was aimed to analyze and study the process of Turmeric fine pulverizing; and the powder properties of Turmeric ultra-micro powder after the process. Based on d50, the powder properties of Turmeric ultra-micro powder were summarized by using orthogonal design to select the optimal Turmeric fine pulverizing. Compari-sons were made on powder properties, such as exterior characters, IR spectra, fluidity and hygroscopicity before and after fine pulverizing. The results showed that optimal fine pulverizing process was determined based on orthogonal design. The conditions were that the material was 1 200 g, with water of 5.5% and crushing for 40 min. Compari-son of powder properties of Turmeric powders before and after fine pulverizing showed that as the diameter of the particle decreased. Turmeric particle gradually showed signs of aggregation. At the same time, granular sensation disappeared;the color turned lighter;powder became finer;fluidity was reduced;balanced hygroscopic capacity el-evated. Although the chemical composition and molecular structure had not changed;initial velocity and capacity of hygroscopicity increased, acceleration declined. It was concluded that Turmeric fine pulverizing was a convenient, reliable and practical process, with small size of particle. It can be used for Turmeric fine pulverizing. The compre-hensive evaluation showed that ultrafine powder four as the optimum powder.

OBJECTIVE: To further define the modulation effect of signal transducers and activators of transcription 3 (STAT3) in adriamycin-resistant breast cancer and to promote the clinical application of the inhibitors of STAT3 in reversing multidrug resistance in cancer. METHODS: Firstly, the levels of STAT3 and phosphorylated STAT3 (pSTAT3) expression in clinical breast cancer tissue samples were determined by Western blotting. The expression of STAT3 and pSTAT3 in adriamycin-sensitive and adriamycin-resistant breast cancer cell lines was evaluated by RT-PCR and Western blotting. Secondly, the expression of STAT3 was detected by Western blotting after blocking the STAT3 signal pathway with small interfering RNA targeting STAT3 (STAT3-siRNA). Finally, after the expression of STAT3 was blocked by STAT3-siRNA, immunofluorescence was performed to study the proliferation activity of breast cancer cells and MTT was used to determine the IC50 of the cells in order to observe whether STAT3-siRNA had any inhibitory effects on the growth of breast cancer cells and whether it could promote the efficacy of adriamycin in breast cancer cells. RESULTS: STAT3 and pSTAT3 were highly expressed both in clinical breast cancer tissue samples and in adriamycin-sensitive-and-resistant breast cancer cells. The expression of pSTAT3 in adriamycin-resistant breast cancer cells was significantly higher than in adriamycin-sensitive breast cancer cells. STAT3-siRNA conspicuously decreased the expression of STAT3 protein and inhibited the growth of adriamycin-sensitive breast cancer cells. Compared with the single application of adriamycin, IC50 of adriamycin-resistant breast cancer cells decreased by 4 fold when adriamycin was used in combinaiton with STAT3-siRNA. Meanwhile, an inhibition of the expression of the anti-apoptotic protein mediated by STAT3 was observed in adriamycin-resistant breast cancer cells. CONCLUSION: This study reveals that the development of breast cancer is related to the activation of STAT3. The activation of STAT3 in adriamycin-resistant breast cancer cells was more notable than in adriamycin-sensitive cells. The inhibition of the STAT3 pathway could improve the adriamycin sensitivity of adriamycin-resistant breast cancer cells and lead to their apoptosis. The RESULTS of this study explores the feasibility of the reversal of drug resistance in cancer by blocking STAT3 pathway and establishes the experimental basis for promoting the clinical use of the inhibitors of STAT3 pathway and the chemotherapeutics to overcome the multidrug resistance in cancer.

OBJECTIVETo observe the inhibitory effect of a eukaryotic expression vector expressing human IFN-gamma (pcDNA3.1- IFN-gamma) on HBV replication in hepG2.2.15 cells.

METHODSThe eukaryotic expression vector expressing human IFN-gamma was constructed using PCR and gene recombination technique. hepG2.2.15 cells were transfected with pcDNA3.1-IFN-gamma and the culture supernatant was collected to determine the expression of IFN-gamma protein by ELISA. The HBV DNA copies and the concentration of HBeAg and HBsAg were measured by fluorescence real-time PCR and ELISA kit, respectively.

RESULTSCompared with that of negative control and blank 2.2.15 cells, the concentration of HBeAg in the supernatant of 2.2.15 cells transfected with pcDNA3.1- IFN-gamma were decreased by 49%, and HBsAg concentration was lowered by 35% and 33%, respectively. A significant decrease of HBV DNA copies was observed in pcDNA3.1- IFN-gamma-transfected cells in comparison with the two control cells. No significant differences were noted in all the results between the two control groups.

CONCLUSIONWe have successfully constructed the eukaryotic expression vector expressing human IFN-gamma, which provides a basis for anti-HBV gene therapy using human IFN-gamma.

Genetic Vectors ; Hep G2 Cells ; Hepatitis B virus ; drug effects ; Humans ; Interferon-gamma ; genetics ; pharmacology ; Transfection ; Virus Replication ; drug effects

The key challenge to generate engineered cells by synthetic biology for producing 7-dehydrocholesterol (7-DHC) in a high titer is the match between functional module and chassis. Our study focused on solving this problem by combining different promoters and yeast chassis to increase 7-DHC production. To optimize the chassis in order to accumulate zymosterol, the substrate for 7-DHC synthesis, we overexpressed truncated HMG-CoA reductase (tHmglp) and squalene epoxidase (Erglp), both are key genes of yeast endogenous zymosterol biosynthetic pathway. In addition, we knocked out C-24 methyl transferase (Erg6p) and C-22 dehydrogenase (Erg5p) to inhibit the conversion of zymosterol to ergosterol. By introducing heterologous C-24 reductase under three promoters with different strengths, namely TDH3p, PGK1p and TDH1p, we constructed functional modules of diverse activities. Nine engineeredcells were generated based on the combination of these three modules and three chassis. The result shows that the engineered cell composed of functional module regulated by TDH3p and chassis SyBE_000956 had the highest 7-DHC production, indicating a better match than others. This study provides evidences for importance of match and empirical support for rational design of subsequent researches.
Cholesterol ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; Dehydrocholesterols ; metabolism ; Gene Knockout Techniques ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Industrial Microbiology ; Methyltransferases ; genetics ; Promoter Regions, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; genetics ; Synthetic Biology