BACKGROUND: There is a great debate but little research addressing the cell suspension obtained from the digested mantle tissues can effective amplify and form pearl sac in vitro, thus producing pearl. OBJECTIVE: To establish an effective technique and method of in vitro separation and culture of mantle of the pearl oyster (Pinctada martensii), and to determine the optimal method of forming pearl sac with the intact structure and secretion function, thus producing pearl. DESIGN, TIME AND SETTING: Single sample observation was performed at the School of Basic Medicine, Guangxi Traditional Chinese Medical University, between August and December in 2008. MATERIALS: Pearl oyster (Pinctada martensii) aged 1.0 2.0 years, were offered by Yingpan Pearl Industrial Co., Ltd. Of Beihai City, China; the self-modified marine shellfish balanced salt solution; the self-prepared concha pteriae serum and concha pteriae body fluid; keratinocyte growth factor was purchased from Sigma, USA. METHODS: The mantle of pearl oyster (Pinctada martensii) was digested with 2.5 g/L trypsin, the harvested cells were cultured using M199 medium containing 10% fetal bovine serum and supplemented with 10 μg/L keratinocyte growth factor, 10% self-prepared concha pteriae serum and concha pteriae body fluid. The cultivation was performed for 30 days. MAIN OUTCOME MEASURES: Cell growth characteristics and growth state. RESULTS: The pearl mantle epithelial cells cultured in vitro were shown to proliferate rapidly, secrete productively, and the muscle cells showed a great proliferation, finally encapsulated the mantle epithelial cells to form pear sac with the intact structure and strong secretion function. CONCLUSION: Using the modified culture technology and culture system, the addition of keratinocyte growth factor can obtain the well growing and secreting pearl sac during in vitro culture of mantle cells.

Objective To investigate the expression levels of interferon-inducible genes (IFIT1,IFIT4,OAS1,OASL,ISG15) in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus(SLE).and the relations between these genes expression levels and disease activity are explored.Methods Sybr green dye based real-time quantitative PCR method was used to detect the expression levels (indicated as-△△Ct value) of WIT1,IFIT4.OAS1,OASL and ISG15 in 76 patients with SJJE and 54 controls.Their expression levels were compared with erythroeyte sedimentation rate (ESR),serum C reactive protein (CRP),complement C3,C4.antinuclear antibody (ANA).anti-double stranded DNA antibody.The associations between the expression levels of IFIT1,IFIT4,OASI.OASL,ISG15,ESR,CRP,complement C3,C4,ANA,anti-double stranded DNA antibody and SLEDAI scores in patients with SLE were analyzed.Results ① The expression levels of WIT1,IFIT4,OAS1,OASL and ISG15 in the SLE patients were significantly higher than those of the normal controls (P＜0.01).The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in active SLE patients were higher than those of inactive SLE patients (P＜0.05).The real time expression levels of IFIT1,IFIT4,OAS1.OASL and ISG15 showed positive correlations with each other (r＞0.5,P＜0.05) in patients with SLE.② The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 were positively correlated with the SLEDAI scores (r＞0.5,P＜0.05).③ There was no correlation between ESR,CRP,complement C3,C4,ANA and the expression levels of IFIT1,IFIT4,OAS1,OASL,ISG15,SLEDAI scores except anti-double stranded DNA antibody (r＞0.5.P＜0.05).Conclusion The expression levels of IFIT1,IFIT4,OAS1,OASL and ISG15 in patients with SLE are significantly higher than those of the normal controls,and positively associated with SLEDAI scores,so they are helpful in evaluating SLE disease activity and severity.IFIT1,IFIT4,OAS1,OASL and ISG15 genes may be the potential treating targets for SLE.

OBJECTIVETo identify genotype of eight strains of Orientia tsutsugamushi (O. tsutsugamushi) isolated from Nan Peng Lie Islands in China and establish tsutsugamushi disease nature foci for this region.

METHODSThe nested polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were used. Three primers were selected from the DNA sequence of the gene encoding type-specific 56-kDa protein of the Karp strain. The positive products were digested by Hine II and Pst I, meanwhile profiles specific to each strain were analyzed.

RESULTSThree genotypes of O. tsutsugamushi including Karp, Kato and a new strain existed on Nan Peng Lie Islands.

CONCLUSIONNan Peng Lie Islands is tsutsugamushi disease nature foci.

Animals ; China ; DNA, Bacterial ; analysis ; Genotype ; Mice ; Orientia tsutsugamushi ; genetics ; Polymorphism, Restriction Fragment Length ; Rats

OBJECTIVETo investigate natural foci of tsutsugamushi disease whose incidence has increased in the Nan Peng Lie Islands in China, an area where this disease has not been previously recorded.

METHODSWe recorded the natural foci and isolated Orientia tsutsugamushi (O. tsutsugamushi) organism. We also studied prevention measures.

RESULTSThese islands had the natural foci of a south subtropical zone. The main host and vector were Rattus norvegicus and Leptotrombidium deliens (L. deliens), respectively. The seasonal quantity trends of Rattus norvegicus and Leptotrombidium deliens were consistent with the incidence of human infection. Thirty-five strains of O. tsutsugamushi were isolated from Rattus norvegicus and L. deliense. The identification of 7 strains showed that most strains were Karp. Seroepidemiology showed a high prevalence of antibody against O. tsatsugamushi among local people. After prevention measures were used, the incidence was decreased.

CONCLUSIONThis was the first successful confirmation that the Nan Peng Lie Islands were natural foci of tsutsugamushi disease.

Animals ; Antibodies, Bacterial ; blood ; Cattle ; China ; epidemiology ; DNA, Bacterial ; genetics ; Geography ; Humans ; Incidence ; Orientia tsutsugamushi ; genetics ; growth & development ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Rats ; Scrub Typhus ; epidemiology ; microbiology ; Seasons ; Sheep ; Species Specificity ; Tropical Climate

Objective There are few researches for the serum betatrophin level and diabetic nephropathy (DN) recently.The aim of this study was to investigate the change of serum betatrophin level and the correlation of serum betatrophin and urinary albumin-to-creatintine ratio (UACR) in patients with type 2 diabetes mellitus (T2DM).Methods A total of 150 Chinese subjects from Mar 2013 to Jul 2016 were enrolled in the study, including 90 patients with type 2 diabetes and 60 healthy controls.According to the level of UACR, the diabetic patients were divided into two groups:normal UACR group (UACR<30 mg/g, n=60) and abnormal UACR group(UACR>30 mg/g, n=30).Serum betatrophin was measured by enzyme linked immunosorbent assay (ELISA).UACR was measured by turbidimetric inhibition immune assay.Blood glucose blood lipid were measured simultaneously.Results The serum betatrophin level was significantly higher in abnormal UACR group than that in normal UACR group[677.37±59.02 vs 486.13±41.22 pg/mL, P<0.05];Serum betatrophin level in T2DM patients was positively correlated with age (r=0.246), waist hip ratio (WHR) (r=0.240), fasting blood glucose (FPG) (r=0.234), 2 hour plasma glucose (2hPG) (r=0.363), glycosylated hemoglobin (HbA1c) (r=0.346), fasting insulin (FINS) (r=0.249), insulin resistance index (HOMA-IR) (r=0.309), blood urea nitrogen (BUN) (r=0.223), creatinine (CREA) (r=0.277) and UACR (r=0.244) (P<0.05),and negatively correlated with glomerular filtration rate (GFR) (r=0.308) (P<0.01).Serum betatrophin level in normal UACR group was positively correlated with age, HbA1c and UACR (P<0.05);Serum betatrophin level in abnormal UACR group was positively correlated with WHR (r=0.504), 2hPG (r=0.600), HbA1c (r=0.449), HOMA-IR (r=0.395) (P<0.05).The WHR, HbA1c, HOMA-IR and GFR were the influential factors of the serum betatrophin level.Conclusion The level of serum betatrophin was significantly increased in T2DM patients with albuminuria, which suggests that the betatrophin might play an important role in the pathogenesis of DN.

Objective:To investigate and analyze the somatic symptom disorder, anxiety and depression in patients with myocardial bridge.Methods:A total of 276 patients with myocardial bridge diagnosed by coronary angiography (CAG) were enrolled in the department of cardiology of Renji hospital in Shanghai from June to December in 2016. There were 151 cases of simple myocardial bridge (no coronary stenosis or coronary artery stenosis <30%) and 125 cases of complex myocardial bridge (combined with coronary stenosis >30%). A total of 1067 patients with myocardial bridge without coronary angiography were collected at the same time. Self-rating somatic symptom scale (SSS), generalized anxiety disorder (GAD-7) and patient health questionnaire (PHQ -9) were given to these patients during hospitalization. At the same time, somatic symptoms disorder and anxiety and depression in the myocardial bridge group and non-myocardial bridge group were compared.Results:The prevalence of somatic symptom disorder in patients with myocardial bridge was higher than that in non-myocardial bridge patients (35.86% vs 28.30%, P<0.05). There was significant correlation between somatic symptom disorder and depression and anxiety, with correlation coefficients of 0.629 and 0.565, respectively. The prevalence of depression and anxiety in myocardial bridge patients was higher than that in non-myocardial bridge patients (depression: 23.91% vs 22.11%. P=0.467; anxiety: 17.02% vs 14.15%, P=0.22), but there was no statistical difference. For male patients or female patients, the prevalence of somatic symptom disorder, depression and anxiety in the simple myocardial bridge patients were higher than those in the complex myocardial bridge patients, but there was no statistical difference. The most common non-specific somatic symptoms disorder in patients with myocardial bridge were fatigue (64.5%), followed by sleep disorders (63.8%) and decreased attention (63.0%). Conclusion:The somatic symptom disorder in patients with myocardial bridge is significantly higher than that in non-myocardial bridge group. Especially for patients with myocardial bridge with non-specific somatic symptoms, early identification of somatic symptoms disorder of myocardial bridge patients will be beneficial to proper clinical invitation.

Background: Replacing damaged anterior cruciate ligaments (ACLs) with tissue-engineered artificial ligaments is challenging because ligament scaffolds must have a multiregional structure that can guide stem cell differentiation. Here, we designed a biphasic scaffold and evaluated its effect on human marrow mesenchymal stem cells (MSCs) under dynamic culture conditions as well as rat ACL reconstruction model in vivo. Methods: We designed a novel dual-phase electrospinning strategy wherein the scaffolds comprised randomly arranged phases at the two ends and an aligned phase in the middle. The morphological, mechanical properties and scaffold degradation were investigated. MSCs proliferation, adhesion, morphology and fibroblast markers were evaluated under dynamic culturing. This scaffold were tested if they could induce ligament formation using a rodent model in vivo. Results: Compared with other materials, poly(D,L-lactide-co-glycolide)/poly(ε-caprolactone) (PLGA/PCL) with mass ratio of 1:5 showed appropriate mechanical properties and biodegradability that matched ACLs. After 28 days of dynamic culturing, MSCs were fusiform oriented in the aligned phase and randomly arranged in a paving-stone-like morphology in the random phase. The increased expression of fibroblastic markers demonstrated that only the alignment of nanofibers worked with mechanical stimulation to promote effective fibroblast differentiation. This scaffold was a dense collagenous structure, and there was minimal difference in collagen direction in the orientation phase. Conclusion Dual-phase electrospun scaffolds had mechanical properties and degradability similar to those of ACLs. They promoted differences in the morphology of MSCs and induced fibroblast differentiation under dynamic culture conditions. Animal experiments showed that ligamentous tissue regenerated well and supported joint stability.

Background: Replacing damaged anterior cruciate ligaments (ACLs) with tissue-engineered artificial ligaments is challenging because ligament scaffolds must have a multiregional structure that can guide stem cell differentiation. Here, we designed a biphasic scaffold and evaluated its effect on human marrow mesenchymal stem cells (MSCs) under dynamic culture conditions as well as rat ACL reconstruction model in vivo. Methods: We designed a novel dual-phase electrospinning strategy wherein the scaffolds comprised randomly arranged phases at the two ends and an aligned phase in the middle. The morphological, mechanical properties and scaffold degradation were investigated. MSCs proliferation, adhesion, morphology and fibroblast markers were evaluated under dynamic culturing. This scaffold were tested if they could induce ligament formation using a rodent model in vivo. Results: Compared with other materials, poly(D,L-lactide-co-glycolide)/poly(ε-caprolactone) (PLGA/PCL) with mass ratio of 1:5 showed appropriate mechanical properties and biodegradability that matched ACLs. After 28 days of dynamic culturing, MSCs were fusiform oriented in the aligned phase and randomly arranged in a paving-stone-like morphology in the random phase. The increased expression of fibroblastic markers demonstrated that only the alignment of nanofibers worked with mechanical stimulation to promote effective fibroblast differentiation. This scaffold was a dense collagenous structure, and there was minimal difference in collagen direction in the orientation phase. Conclusion Dual-phase electrospun scaffolds had mechanical properties and degradability similar to those of ACLs. They promoted differences in the morphology of MSCs and induced fibroblast differentiation under dynamic culture conditions. Animal experiments showed that ligamentous tissue regenerated well and supported joint stability.