Objective To determine the genetic subtypes and susceptible population of Cryptoccocus neoformans in China. Methods We analyzed 129 strains of Cryptococcus neoformans isolated during the period from 1980 through 2006 in 18 Prov-inces and municipalities of central and eastern China using epidemiological method,PCR fingerprint,and multilocus sequence typing (MLST).Results About 71.5% (91/129)of the clinical strains were isolated from the patients without any recognizable immunodeficiency.Only 8.5% (11/129)of the strains were from AIDS patients.The remaining 20.9% (27/129)were from patients with underlying diseases other than HIV infection.Furthermore,PCR fingerprinting and MLST showed that serotype A strains exhibited a unique VNⅠsubtype,which was distinguishable from the reference VNⅠmolecular types.For convenience we named this unique genotype as VNⅠc.Conclusions People without any recognizable immunodeficiency are the main susceptible population of the 129 clinical strains of Cryptococcus neoformans ,which contrasts with the reports from other countries.The different genetic subtypes of Cryptococcus neoformans between China and other countries may contribute to such a difference.

Polypoidal choroidal vasculopathy(PCV) is a common disease of the retina,the disease is more found in colored race.In China,the mainly nosogenic population is the female about 60 years old.The pathogenesis of PCV is unknown and there is no specific medicine at present.The funduscopic present is orangered lesions,bleed and exudation.Indocyanine green angiography(ICGA) could make a definite diagnosis of PCV.In recent,the main treatment of PCV includes photodynamic therapy,intravitreal anti-VEGF therapy,surgery for treatment of complications and combined therapy.We review the treatment and diagnosis of PCV,and propose the further research direction.

We investigated the Toxop lasma infection prevalence among planned pregnant women in Chongqing ,and to pro‐vide reference for the first level intervention of birth defects in the region .A total of 11 953 planned pregnant women were se‐lected by proportionally stratified multi‐stage random sampling method .Questionnaire survey was given to the women ,and blood samples were collected .Specific IgM and IgG antibodies against Toxoplasma were detected with ELISA .Results showed that among the 11 953 cases surveyed ,Toxoplasma IgM was positive in 71 cases ,with the positive rate of 0 .59% ;IgG was positive in 771 cases ,with the positive rate of 6 .54% .The positive rate of IgM and IgG antibodies in the metropolitan core re‐gion of the city was higher than that in the suburb areas (χ2 =35 .28 ,P<0 .000 1 ;χ2 =82 .65 ,P<0 .000 1) .The positive rate of IgM antibody increased with the educational level (χ2trend=3 .25 ,P=0 .001 1) .The positive rates of IgM and IgG varied in occupations among women (χ2 =13 .93 , P= 0 .016 0;χ2 = 15 .58 ,χ2 =0 .008 1) ,with the highest rate of public officials . Planned pregnant women with the history of abnormal pregnancy outcomes had higher positive rate of T .gondii IgM and IgG antibodies than those in the control (χ2 =6 .85 ,P=0 .008 9;χ2 =59 .25 ,P<0 .000 1) .There was no significant difference of IgM positive rate between the planned pregnancy women who had closed contact to cats and the control group (χ2 =0 .23 ,P=0 .628 6) ,while the positive rate of IgG was higher than that of the control group (χ2 =9 .95 ,P=0 .001 6) .T .gondii infec‐tion rate was on the low level of planned pregnant women in Chongqing .Adverse pregnancy outcomes are related to Toxoplas‐ma infection .

Adolescent ; Female ; Humans ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; injuries ; physiopathology ; surgery ; Range of Motion, Articular ; Reconstructive Surgical Procedures

Objective To study the applications and outcomes of using autogeneic cartilage in hearing recon-struction surgery in patients with chronic otitis media or cholesteatoma .Methods A total of 165 patients (173 ears) in whom autogeneic cartilage was used were analyzed retrospectively .Forty -three patients (48 ears) had simple tympanic membrane perforations ,61 patients (61 ears) had cholesteatomas including 12 retraction pockets ,23 pa-tients (23 ears) had tympanoscleroses and 38 patients (41 ears) had otitis media with granulations .The cartilage grafts were used for tympanic perforation reparing in 133 patients (139 ears) ,for ossiculoplasty in 102 patients (104 ears) ,for attic reconstruction in 31 patients (31 ears) and for canal wall reconstruction of external auditory canal in 3 patients (3 ears) .The auditory outcome (0 .5 ,1 ,2 ,and 4 kHz pure tone average hearing threshold ,the average air-bone gap) and local architecture status were followed up for 1 year after surgery .Results In 133 patients (139 ears) with tympanic perforation ,the rate of successful repair of a tympanic membrame perforation in one -stage was 97 .84% with perforation repair in 136 ears and postoperative perforation in 3 ears .In 102 patients (104 ears) of os-siculoplasty ,there was no ossicular prostheses prolapse .In 31 patients (31 ears) of attic reconstruction ,no local graft shift or collapse was found .In 3 patients (3 ears) of external auditory canal repair ,no canal wall collapse occurred . In myringoplasty group (43 patients ,48 ears) ,preoperative and postoperative air -bone gap (ABG) was 23 .8 ± 3 .1 dB and 11 .6 ± 8 .7 dB ,respectively .In cholesteatoma group (61 patients ,61 ears ) ,preoperative and postoperative ABG were 39 .2 ± 24 .7 dB and 19 .0 ± 12 .1 dB ,respectively .In tympanosclerosis group (23 patients ,23 ears) ,pre-operative and postoperative ABG were 31 .2 ± 12 .4 dB and 19 .8 ± 11 .2 dB ,respectively .In otitis media with granu-lation group (38 patients ,41 ears) ,preoperative and postoperative ABG were 41 .6 ± 9 .9 dB and 15 .3 ± 13 .4 dB ,re-spectively .Conclusion Autogeneic cartilage is very valuable in hearing reconstruction surgery ,especially in compli-cated tympanic perforation ,combination with ossiculoplasty prostheses ,or reconstruction of mastoid cavity or exter-nal call wall defect .

Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.

Objective To express and purify recombinant Tp0136 epitope fragment, and study the immunity activity. Methods The Tp0136 selective fragment(Tp0136B) gene was devised by the surface property analysis, solvent-accessible suface calculateions, secondary structure function region analysis, and was inserted between the sites of Nde Ⅰ and Not Ⅰ in pET22b ( + ) . The recombinant plasmid was expressed in E. coli BI21. After nickel ion metal affinity chromatography, the antigenic and immune reactivity of rTp0136B was confirmed. Then indirect ELISA with the rTp0136B as coating antigen was performed to detect the anti-Tp0136 antibody in sera from 100 normal human controls and 131 primary syphilis patients. Results The rTp0136B was soluble expressed with a molecular weight of about 28 000 and was obtained with a purity of ＞98% by chromatography. Western blot proved that the rTp0136B could specifically react with anti-Tp0136 polyclonal antibody. Specific humoral response was elicited by the recombinant protein in Japan negative. The positive detection rate in sera from primary syphilis patients was 85.5%. Conclusion This result suggested that the recombinant Tp0136 epitope fragments have a satisfactory immunocompetence,which may have applications in the serodiagnosis of primary syphilis.

Objective To establish ultra-high performance liquid chromatography-evaporative light scattering detector (UPLC-ELSD) method to determine the content of astragaloside in Guiqi Yiqi Yangxue Mixture and optimize the best water decoction process of the mixture.Methods The UPLC-ELSD conditions to determine the content of astragaloside in the mixture were as follow.The method was carried out on a Waters Acquity UPLC BEH C18 column (2.1 mm ×50 mm,1.7 μm) by using acetonitrile-water (35∶65) as mobile phase, and the flow rate was 0.3 mL/min and the column temperature 30℃.The parameter of ELSD:the drift tube temperature was set at 60℃, and the gas flow rate of N2 2.0 L/min.The content of paeoniflorin in the mixture was detected by previously established HPLC method.The orthogonal test was designed to optimize the water decoction process of the amount of water (A), decoction time (B) and decoction times (C), and the extract yield and the contents of astragaloside and paeoniflorin were evaluated as the comprehensive markers for the purpose.Results The UPLC-ELSD determination of astragaloside had a good linear relationship (r =0.9999), its mean recovery was 103.0%, and it had a good precision, specificity and repeatability.The orthogonal experimental results showed that decoction times (C) >the amount of water (A) >decoction time (B) were major factors affecting the decoction process in order and the ANOVA result showed that decoction times has significant effect on the decoction process (P<0.05).The optimal process of Guiqi Yiqi Yangxue Mixture was adding eight-fold water each time to decoct twice for 1.5 h totally.Conclusion This optimized water decoction process is feasible and stable, and suitable for industrialized production.

Objective To expore the difference between magnetic resonance image ( MRI) expression of spinal tuberculosis and brku-cellare spondylitis. Methods Retrospectively analyzed the clinical data of 10 patients with tuberculous spondylitis and 12 patients with bru-cellar spondylitis from Jan. 2012 to Oct. 2013. All the patients were scanned by MRI, and the expression difference of MRI were compared. Results The vertebral body of spinal tuberculosis was destroyed severely, and it often accompanied by the kyphosis and multiple-level para-vertebral abscess, and even adjacent organs tuberculosis. The vertebral body of brucellar spondylitis was destroyed lightly,and the abscess is often limited. Conclusion We can distinguish spinal tuberculosis and brucellar spondylitis by the typical difference of the expression of MRI.

Aim To investigate the effects of leptin on ?1(Ⅰ) collagen mRNA expression and protein production, and the roles of Janus kinase/signal transducers and activators transcription(JAK/STAT) signaling transduction pathway in increased ?1(Ⅰ) collagen mRNA expression stimulated by leptin in activated hepatic stellate cells(HSCs).Methods Firstly, ?1(Ⅰ) collagen mRNA expression and protein production as well as JAK1 and STAT3 phosphorylation induced by leptin at different doses in a human HSC cell line, LX-2 were determined by RT-PCR, ELISA, and Western-blot.Secondly, the effects of JAK1 inhibitor AG490 on JAK1 phosphorylation and ?1(Ⅰ) collagen mRNA expression stimulated by leptin were detected by Western blot and RT-PCR. Thirdly, the roles of AG490 and transfection with STAT3 antisense oligonucleotide(STAT3-ASON) in STAT3 phosphorylation after leptin were detected by Western blot. Finally, the effect of transfection with STAT3-ASON on ?1(Ⅰ) collagen mRNA expression after leptin was measured by RT-PCR.Results Leptin increased ?1(Ⅰ) collagen mRNA expression and protein production in a dose-dependent manner in LX-2, reaching a maximal level at 80 ?g?L-1 leptin. In addition, phosphorylation of JAK1 and STAT3 after leptin exhibited a time-dependent effect. Besides, JAK1 inhibitor AG490 completely blocked JAK1 and STAT3 phosphorylation and increased in ?1(Ⅰ) collagen gene expression after leptin in LX-2. Transfection with STAT3-ASON blocked STAT3 phosphorylation and increased ?1(Ⅰ) collagen mRNA by leptin in LX-2.Conclusion Leptin had a direct action on liver fibrogenesis by stimulating ?1(Ⅰ) collagen mRNA expression and protein production in activated HSC, and JAK/STAT signaling transduction pathway was involved in the process. JAK1 inhibitor AG490 and transfection with STAT3-ASON blocked the transduction pathway effectively in LX-2. Leptin may be an important factor in the development of hepatic fibrosis. Activated JAK1 and STAT3 signaling in human HSC line provided a novel molecular target for therapeutic intervention of liver fibrosis.