Objective To obtain the recombinant urease B subunit (ureB) of Helicobacter pylori (Hp) and apply it in serological detection of Hp infected patients. Methods Urease B subunit gene was amplified from the complete genome of Helicobacter pylori by PCR, and cloned into the PinPoint TM Xa Ⅲ fusion expression vector, then was sequenced. The protein of urease B subunit was expressed in E.coli JM109 and purified with affinity chromatography. 113 serum samples of peptic ulcer patients were detected by ELISA combined with purified UreB protein. Results DNA sequence analysis showed the nucleic acid sequence homology of ureB gene was 96.44% and the putative amino acid sequence homology was 99.65%. The recombinant UreB protein was composed of 571 amino acid residues and kept original immunologic reaction with corresponding antibody, and its purity was over 90% after affinity chromatography. The results of ELISA associated with recombination UreB antigen showed the sensitivity and specificity was 92%, 98.5%. Conclusion The recombinant UreB protein will be of value for clinical serodiagnosis and epidemiological study of Helicobacter pylori.

Objective To investigate the relationship between the expression of Toll-like receptor 4 (TLR4) on mononuclear cells in peripheral blood and changes of serum vitamin D level in children with bronchiolitis.Methods The children who were diagnosed as bronchiolitis and received treatment in the Second Affiliated Hospital of Harbin Medical University from October 2013 to January 2014 were chosen as the pre-treatment group,and then divided them into moderate group and severe group according to the clinical symptoms,20 cases for each group.Then the cases in pre-treatment group who recovered after treatment were recruited as the after-treatment group,and the children who were healthy and medical examination in the Second Affiliated Hospital of Harbin Medical University in the same period were recruited as the healthy control group.The expressions of TLR4 on CD14 labeled mononuclear cells in the periphera were measured by flow cytometry.The level of 25 (OH) D in serum was detected by enzyme linked immunosorbent assay (ELISA).Results (1) The expression level in children with bronchiolitis of TLR4:the mode-rate group [(18.98 ±2.29)％] and severe group [(30.13 ±2.13)％] increased significantly (P ＜0.05) compared with control group [(1.17 ± 0.57) ％].And the expression level of moderate group [(2.02 ± 0.48) ％] and severe group [(11.43 ± 1.52) ％] decreased significantly after treatment (P ＜0.05).(2) Serum vitamin D level in children with bronchiolitis of the moderate group[(17.16 ± 3.34) μg/L] and severe group [(6.56 ± 2.28) μg/L] were lower than healthy control group [(53.69 ± 20.18) μg/L] before treatment (P ＜ 0.05),especially the severe group [(6.56 ±2.28) μg/L].The level of moderate group [(9.59 ± 2.31) μg/L] and severe group [(4.70 ± 0.67) μg/L] became lower after treatment (P ＜ 0.05).(3) Both severe group (r =-0.491,P ＜ 0.05) before treatment and moderate group (r =-0.436,P ＜ 0.05) after treatment showed negative correlation between TLR4 on mononuclear cells in peripheral blood and serum 25 (OH)D level in children with bronchiolitis.And no correlation was found among healthy control group,moderate group before treatment and severe group after treatment (P ＞ 0.05).Conclusions The conditions of children with bronchiolitis was positively correlated with the expression level of TLR4,and negatively correlated with the vitamin D level.The serum 25 (OH) D decreased steadily during the treatment.The expression of TLR4 in monocytes has a certain correlation with the level of vitamin D in children with bronchiolitis.

Objective:To analyze the polymorphism of urease B gene(ureB) from different Hp strains. Methods:The sequences of ureB gene from 14 different Hp strains were amplified by PCR to analyze PCR-RFLP with HaeⅢ digestion.At the same time,the sequence of ureB gene was analyzed with biochemical software to compare the homology of nucleotide and amino acid sequence and to profile the phylogenetic tree. Results: The results of 1.7 kb ureB gene digested with HaeⅢ showed there were 2-5 band types in 14 different Hp strains and formed five distinct RFLP types.The two reference strains had the same RFLP type as 3 clinical isolates while the other clinical isolates and 3 isolates from animal model belong to 4 different RFLP types respectively.The nucleotide sequences and putative amino acide sequences of Hp ureB were compared.The various ureB sequences had high homologies(more than 96.6% and 98% in nucleotide sequences and amino acide sequences respectively) among 14 Hp strains.Particularly,there was the highest homologies(100%) between CCS9801、CCS9806、M3 and M10.Phylogenetic tree analysis showed that two reference strains and other isolates from clinical patients and Hp-infected mice model were located in two different lineage respectively in phylogenetic tree of nucleotide sequences while there were some variance in phylogenetic tree of amino acide sequences. Conclusion: Hp ureB was high conserved and homologous in the gene level as well as in the protein level.

Objective To assess the biophysical characteristics of normal skin and inflammatory skin lesions in mild and moderate acne. Methods Seventy-five mild and moderate acne vulgaris patients were included in the study. One inflammatory lesion measuring 2.0 to 5.0 mm in diameter which occurred within 48 hours prior to the measurement was selected as the target lesion. Trans-epidermis water loss (TEWL), capacitance and a* value were measured in target lesions and lesion-adjacent normal skin. Sebum content immediately,1 and 4 (saturated) hours after face washing was also determined on target lesions and central forehead between the eyebrows. Sebum secretion rate (SSR) was calculated. Results The TEWL and SSR significantly increased in the lesion-adjacent normal skin with the increment of inflammatory lesion number (both P < 0.05), and increased in target lesions with the elevation of a* value (both P < 0.05). The target lesions exhibited a significantly higher TEWL but a lower saturated sebum content and SSR than the adjacent normal skin did (all P <0.05). Conclusions The severity of inflammatory lesions is correlated with the impaired barrier function and increased SSR in facial skin. Compared with the adjacent normal skin, inflammatory skin lesions have a reduced skin barrier function and SSR.

Objective To evaluate the efficacy and safety of total knee arthroplasty (TKA) with 3D printing guild plate by comparing with conventional TKA. Methods From May 2014 to September 2014, 40 patients suffered primary unilateral TKA were received, in which there were 11 males and 29 females, aged from 57 to 82 years with an average age of 68.5±6.3 years. The subjects were divided into two groups randomly. One group was treated with TKA with 3D printing guild plate while the other group was treated with the conventional TKA. The blood loss, operation time, post?operative Hospital for Special Surgery (HSS) score, range of motion (ROM), lower limb mechanical alignment and incidence of complication were compared with insignificant differences. Results The operation time in the 3D printing TKA group (103.4±11.7 min) was lower than that in the conventional TKA group (124.5±21.6 min), which was statistically significant (t=3.838, P=0.000). The blood loss in the 3D printing TKA group (370.2±96.0 ml) was lower than that in the conventional TKA group (510.0±235.9 ml), which was statistically significant (t=2.454, P=0.019). The post?operative ROM of knee in the 3D printing TKA group was 104.3° ± 15.5° and that in the conventional TKA group was 103.5° ± 12.5° (t=0.169, P=0.867). HSS scores in the 3D printing TKA group and in the conventional TKA group were 88.5±5.7 and 89.4±4.8, which was statistically insignificant (t=-0.633,P=0.530). Mechanical alignment in the 3D printing TKA group was 2.9°±1.1° and that in the conventional TKA group was 3.0°±0.9°, which was not statistically significant (t=-0.317, P=0.753). No obvious complication occurred in two groups. Conclusion TKA with 3D printing guild plate has similar results to conventional TKA in HSS score, mechanical alignment and ROM of knee, but it shortens operation time and decreases the blood loss.

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.

AIM:To investigate the cause of blindness, except those caused by cataract, in Haimen city.METHODS:According to the WHO`s criteria of blindness, the blindness level was decided through ophthalmic tests by associate chief or chief ophthalmologists who were trained especially for disability evaluation.The analysis of the the leading cause were taken too.RESULTS:Totally 3 266 persons were blindness, in which 2 118 were first level blindness, 1 148 persons were second lever blindness, and 1 308 persons were male, 1958 were female.The leading cause of blindness were retina and uveitis diseases (31.58%), genetic diseases(23.47%), cornea disease(14.49%).CONCLUSION:The leading cause of blindness are retina and uveitis diseases, genetic diseases, cornea diseases in Haimen city of Jiangsu province.Early prevention and treatment should be strengthened to reduce the occurrence of blindness.

A FatB unigene was obtained from the transcriptome dataset of Lonicera japonica Thunb. Full-length FatB cDNA was cloned from buds of Lonicera japonica Thunb., Lonicera japonica Thunb. var. chinensis (Wats.) Bak., Lonicera hypoglauca Miq. and Lonicera dasystyla Rehd. using RT-PCR technology, and named as LJFatB, LHFatB, LJCFatB and LDFatB. The results of bioinformatic analysis showed that LJFatB, LJCFatB, LHFatB and LDFatB and Arabidopsis thaliana AtFatB had a closely relationship. Nucleotide sequences and protein secondary structure of LJFatB, LJCFatB, LHFatB and LDFatB are different and their proteins had conserved FatB substrate binding sites and catalytic activity sites. Transcriptive level of LJFatB, LJCFatB, LHFatB and LDFatB in bud was not significantly different. Therefore, LJFatB, LJCFatB, LHFatB and LDFatB could have the same biological function as AtFatB.

Objective To study the plasma biochemical indexes among the hypertension in Xining area.Methods According to diagnosis standard of hypertension,104 males who emigrated to Xining area were divided into hypertension group and control group,height,weight,blood routine,renal function,blood lipid of the two groups were measured.Results Among the 104 subjects, 17 cases were hypertensive patients,and other 87 cases were served as control group.The body weight,blood uric acid levels in hy-pertension group were significantly higher than those in the control group(P <0.05),the other indexes had no significant difference between two groups.Conclusion The uric acid may be a risk factor for hypertension people living in plateau area,and the mecha-nism need to be further studied.

Objective To analyze plague epidemic tendency in the Three-River Region of Qinghai.Methods Using retrospective study,the Three-River Region during 1954-2006 year pestis epidemic focus were investigated and analyzed.Result Pestis prevailed mainly in Yushu,Chindu,Qumalai,Nangqian,Zhiduo and the Geermu.Tanghla Township.It was first found that the nature plague focus of miefitus existed in Chengduo County.There are 1 5 kinds of 12 branches in 8 trees infected plague animals were founded,336 Yersinia pestis were separated from the driven objects.Among them there were 291 Himalayas marmot body,account for 86.60%of the total,13 of Tibet sheep,accounts for 3.87%.10 of Qinghai field-mouse,accounts for 2.98%,Also there were 114 Yersinia pestis which were separated from each kind of vector insect in vivo.And,46 pestis strains came from the axe shape of flea in vivo account for 40.35%(46/114),38 pestis strains separated from Xie mountain flea,account for 33.33% (38/114).During 1960-2006 years there were 85 human plague cases were founded,238 occurred,134 died,the case fatality rate wero 56.30%(134/238),the popular seasons were started from May to November,the peak season happened in Aug and Sep.After Oct mainly due to Tibet sheep pestis which will cause as the origin of infection.The majority of sickness was pulmonary plague,account for 49.58%(117/238),whereas the first round case caused by the gland bubonic plague,account for 77.12%(91/118).Conclusions There are two pestis strains natural epidemic focus places in Three-River Source Region of Qinghai including the Himalayas marmot pestis strain and the Qinghai field-mouse pestis strain.The case of human pestis strain causes by the marmot strain,the fiehl-mouse mold mushroom spawn causes human pestis strain has not yet discovered,Three-River Source Region of Qinghai is a pestis strain key popular area in Qinghai Province.