Ocular surface tissues are vulnerable to various stimuli from the external environment,including microbial invasion,mechanical damage,chemical stimulation,dust,UV light,and allergen stimulation.Conjunctival goblet cells play an important role in maintaining ocular surface self-homeostasis and health.These cells are the only unicellular glands in humans and mammals that are mainly distributed in the epithelium of the digestive tract,respiratory tract,and the ocular surface.Many factors affect goblet cell development and differentiation,especially the newly discovered molecule Spdef,which is a transcription factor found in the ETS family.The main function of conjunctival goblet cells is to secrete mucin to protect and lubricate the ocular surface;however,they also secrete a variety of other bioactive substances.Recent studies have demonstrated that goblet cells are involved in antigen presentation,regulation of dendritic cell phenotype differentiation,and induction of immune tolerance on the mucosa.Many ocular surface diseases,such as dry eye,will alter density and function of conjunctival goblet cells.Therefore,ophthalmologists should be concerned about the biological behavior of conjunctival goblet cells and the relationship between them and ocular surface diseases.

Objective: To obtain the 12-mer phage clones displaying the Hantaan virus mimic epitopes.Methods: HFRSV-positive sera were used as selective molecules for biopanning.A 12-mer phage peptide library was bio-panned for 5 rounds,and the result was confirmed by sandwich ELISA,competition ELISA and DNA sequencing.Results: After 5 rounds of effective screening,the results detected by sandwich ELISA and competition ELISA showed that the majority of the selected clones could react to positive sera in a dose-dependent manner,but could not bind to BSA and the control sera.Sequencing and alignment analyses indicated that amino acid sequences of 45 positive clones fell into 8 groups,and 7 of them exhibited putative motifs: LVXKR,LTXR,IXKP,LXPA,VGA,KXIR and EKXP.Four of the putative motifs had a homologous region within the structural proteins of HFRSV.Conclusion: The peptides displayed by the phage can mimic the epitopes of HFRSV antigens,which provides the potential for preparing more effective epitope-based vaccines and specific diagnostic reagents.

Objective To observation the effect of treating wind-cold type periarthritis of shoulder with acupuncture with warmed needle. Methods Study the influence of acupuncture with warmed needle on clinical indexes as total therapeutic effect, pain, and range of activity of 60 patients with wind-cold type periarthritis of shoulder. Result The total effective rate was over 80%. Conclusion acupuncture with wanned needle is a good method in treating wind-cold type periarthritis of shoulder clinically.

Objective: To clone,identify and express the Shigalike toxin 2B(Stx2B) subunit gene of EHEC O157∶H7.Methods: A pair of primers were designed based on the Stx2B subunit gene sequence of EHEC O157∶H7.The Stx2B gene was amplified from the EHEC O157∶H7 chromosome by PCR and cloned into the pMD18-T vector.Thereafter,the gene was cut from the pMD18-T vector and cloned into the prokaryotic expression plasmid pET-28a vector.Then the recombinant plasmids were transformed into E. coli BL21(DE3) and the transformed host strain induced by IPTG.The expression protein was detected by SDS-PAGE and Western-blot analyses.Results: The Stx2B gene was successfully cloned into pMD18-T and pET-28a vectors,and the expressed protein identified by SDS-PAGE and Western blot.The molecular mass(Mr) of the expressed product was about 7 500,and the expression rate about 40%. Conclusion: The Stx2B gene was successfully cloned and effectively expressed in the prokaryotic expression system.

The mrp gene of SS2 human strain Habb was truncated and cloned into a prokaryotic expression vector pGEX4T-2,and a fusion-expressed protein MRP-GST of 61 000 was obtained in E.coli.The GST was cut from MRP-GST with thrombin protease to gain the purified MRP of 35 000 which showed a strong reaction to the SS2 positive sera in Western blotting.BALB/c mice were immunitied intraperitoneally with purified MRP protein.Murine myeloma cells were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA coated with purified MRP was used to screen hybridomas for production of specific antibody.The six McAb recognized MRP specially.According to the results of 71 standard strains (48 SS and 34 SS2)by Sandwich ELISA,the coincidence rate was 97.2%,but for 34 SS standard serotype was 100 %.The ELISA method has a potential value for clinical and epidemiological applicationgs.

OBJECTIVE To compare and analyze the drug resistance of Chryseobacterium meningosepticum which producing metallo-?-lactamase(MBL) and extended-spectrum-?-lactamases(ESBLs) from ICU patients′ and non ICU patients′ specimens of sputa so as to guide the rational application of antibiotics.METHODS Identified the strains with VITEK 32,MBL and ESBLs were also screened by double disk synergy;the antimicrobial sensitivity of clinical isolates was tested by VITEK GNS143 and the antimicrobial sensitivity was added and tested by K-B method.RESULTS As a result the rate of producing MBL of C.meningosepticum was 49.0% from ICU patients′ specimens of sputa which was higher than the rate of 13.8% from non ICU patients′;the rate of producing ESBLs of C.meningosepticum was 37.2% from ICU patients′ specimens of sputa which was higher than the rate of 30.6% from non ICU patients′;the rate of drug resistance to AMP/SUB,TZP,CEP,CIP,and LEV from ICU patients′ was higher than that from non ICU patients.CONCLUSIONS Why the high resistance rate of C.mengingosepticum in ICU patients′ specimens of sputa might be due to the high producing ?-lactamases(MBL and ESBLs).

Objective:To observe the influence of multi-disciplinary cooperation and optimization management mode on the negative situation and rehabilitation effect of patients with affective disorder after coronary heart disease and cerebrovascular disease.Methods:A total of 188 patients with emotional disorders caused by coronary heart disease and cerebrovascular disease from January 2017 to December 2018 to our hospital were selected. According to the random number method, 94 cases in the control group adopted the conventional management model, and 94 cases in the experimental group adopted the multidisciplinary collaborative optimization management model. The negative emotions [evaluated by Self-rating Anxiety Scale (SAS), Self-rating Depression Scale(SDS) ] scores before and after management, the coping style (Jalowies) scores, and quality of life [evaluated by World Health Organization Quality of Life-Brief (WHOQOL-BREF) ] scores were compared between the two groups. The clinical rehabilitation effect and satisfaction rate of related management between the two groups were compared.Results:There were no significant differences between the two groups before management of SAS scores, SDS scores, Jalowies scores, and WHOQOL-BREF quality of life scores ( P> 0.05). After management, the SAS scores and SDS scores of the two groups were significantly reduced, and the Jalowies scores and WHOQOL-BREF quality of life scores were significantly increased. After management, the SAS score and SDS score of the experimental group were 47.32±5.68, 49.93±6.49 and 54.95±6.59, 55.33±8.30 in the control group, the difference was significant between two groups ( t value was 8.503, 4.969, P<0.01). The Jalowies scores and WHOQOL-BREF scores of the experimental group were higher than those of the control group ( t value was -27.662--4.290, P<0.01). The total effective rate of rehabilitation in the experimental group was 98.94%(93/94), which was significantly higher than 90.43%(85/94) in the control group, the difference was statistically significant ( χ2 value was 6.760, P <0.05). The total satisfaction rate of the experimental group for management intervention was 100.00%(94/94), which was significantly higher than the total satisfaction rate of the control group, 91.49%(86/94), the difference was statistically significant ( χ2 value was 6.397, P <0.05). Conclusions:In the patients with affective disorder caused by coronary heart disease and cerebrovascular disease, multi-disciplinary cooperation and optimization management mode can significantly reduce the negative emotions of patients after management, improve the way of patients to deal with the disease, effectively improve the clinical rehabilitation effect and the quality of life of patients, improve the satisfaction rate of patients with management, shorten the distance between nurses and patients, and achieve ideal results.

Objective To investigate the status of Helicobacter pylori and “Helicobacter macacae” infection in rhesus and cynomolgus monkeys in China.Methods With the use of 16S rRNA specific primers for Helicobacter spp and Helicobacter pylori ( HP ) from published literatures, and new 16S rRNA specific primers designed for “Helicobacter macacae” ( HM ) , we investigated the infection status of these two Helicobacter spps in both of 45 rhesus and 90 cynomolgus monkeys by qPCR or conventional PCR on stool samples.Results All three primer sets for 16S rRNA exhibited excellently sensitivity and specificity.Both the infection rates of HP and HM were 100% among 45 young adult rhesus monkeys.The infection rate of HP and HM in 90 young adult cynomolgus monkeys were 100% and 97.8%, respectively.Conclusions Helicobacter pylori and “Helicobacter macacae” are present in almost every artificially bred adult rhesus and cynomolgus individuals which may adversely affect the health of laboratory monkeys and the accuracy of related animal experiments.

OBJECTIVE: To investigate gene-therapy for human hepatocellular carcinoma with adenovirus vectors by double suicide gene CD/TK. METHODS: Double suicide gene CD/TK was liberated from eukaryotic vectors pCEA-CD/TK and subcloned into shuttle vectors, and the transfer plasmid pAdtrack-CMV-CD/TK was formed after linearizing with Pac 1. It was recombinated with pAdeasy-1 in bacteria BJ5183. The identified adenovirus plasmid was digested by Pac1 and was transfected into 293 cells to pack the adenoviruses. After PCR determination, its titre was measured, and the infection rate and efficacy were tested in human hepatocellular carcinoma cells. RESULTS: pAdtrack-CMV-CD/TK and pAd-CD/TK were tested by endonuclease digestion. Ad-CD/TK was produced in 293 cells, and the human hepatocellular carcinoma cells (SMMC7721) infected by Ad-CD/TK were killed after 5-FC was used, and bystander effects were observed. CONCLUSION: Recombinant adenoviruses with CD/TK fusion suicide gene have a high infection rate and efficacy for human hepatocellular carcinoma cells.

Objective: To develop Epstein-Barr virus(EBV)-transformed human peripheral blood B cell lines from healthy volunteers of type B.Methods: B lymphocytes from healthy volunteers of type B capable of producing anti-A antibodies were transformed by EBV,while Cyclosporine A(CsA),as an immunosuppressive agent that could selectively inhibit T-lymphocytes and protect B-lymphocytes from regression,was used in the experiment.Then,the supernanant of the cell culture medium was tested with red blood cells of type A,B and O by agglutinin assay.Results: Twelve of the15 EBV-transformed B lymphocyte cell lines were established,with a success rate of 80%,while 9 human B lymphocyte lines secreted anti-A antibodies.Conclusion: Human B lymphocyte lines secreting antibodies to A antigens were successfully developed,which helps further studies of human blood specific monoclonal antibodies.