Changes and significance of serum Klotho protein levels in different stages of chronic kidney disease / 中国综合临床

Objective To explore the changes and significance of Klotho protein levels in different stages of chronic kidney disease (CKD). Methods From March 2015 to December 2017,176 patients with CKD admitted to nephrology department of Inner Mongolia People＇s Hospital were selected as the study object (CKD group), and 80 healthy patients in our hospital were selected as the control group in the same period. The serum Klotho levels of CKD patients and control group at different stages were detected by double antibody sandwich ELISA, and the differences between each group were compared. Results The serum Klotho level of CKD group (( 4. 84 ± 1. 87) μg／L) was significantly lower than that of the control group ((9. 11± 3. 14) μg／L) ( t＝ 13. 82, P＜0. 01) . One-way anova showed that estimated renal glomerular filtration rate (eGFR),serum albumin (ALB),hemoglobin ( Hb),blood calcium ( Ca) and serum Klotho were gradually decreased,while phosphorus (P) and creatinine (Cr) in serum were gradually increased,and the difference was statistically significant among the five stages( all P＜0. 01). Spearman correlation analysis showed that Klotho level was positively correlated with eGFR and Ca,and negatively correlated with CKD stage,Cr and P (r＝0. 369,0. 160,－0. 200,－0. 250,－0. 230,all P＜0. 05). The multiple linear regression equation showed that Klotho level was positively and independently correlated with eGFR ( t＝ 3. 89, P＜0. 001),and negatively correlated with CKD staging independently (t＝－4. 12,P＜0. 001). Conclusion The expression level of serum Klotho protein in patients with CKD is lower than that of healthy people,and it decreases with the increase of CKD stages,which is closely related to the deterioration of renal function. It can be used as a reference index to evaluate the incidence and severity of CKD.

Prevalence of Hymenolepis nana and H. diminuta from Brown Rats (Rattus norvegicus) in Heilongjiang Province, China

Hymenolepis nana and Hymenolepis diminuta are globally widespread zoonotic cestodes. Rodents are the main reservoir host of these cestodes. Brown rats (Rattus norvegicus) are the best known and most common rats, and usually live wherever humans live, especially in less than desirable hygiene conditions. Due to the little information of the 2 hymenolepidid species in brown rats in China, the aim of this study was to understand the prevalence and genetic characterization of H. nana and H. diminuta in brown rats in Heilongjiang Province, China. Total 114 fecal samples were collected from brown rats in Heilongjiang Province. All the samples were subjected to morphological examinations by microscopy and genetic analysis by PCR amplification of the mitochondrial cytochrome c oxidase subunit 1 (COX1) gene and the internal transcribed spacer 2 (ITS2) region of the nuclear ribosomal RNA gene. In total, 6.1% (7/114) and 14.9% (17/114) of samples were positive for H. nana and H. diminuta, respectively. Among them, 7 and 3 H. nana isolates were successfully amplified and sequenced at the COX1 and ITS2 loci, respectively. No nucleotide variations were found among H. nana isolates at either of the 2 loci. Seventeen H. diminuta isolates produced 2 different COX1 sequences while 7 ITS2 sequences obtained were identical to each other. The present results of H. nana and H. diminuta infections in brown rats implied the risk of zoonotic transmission of hymenolepiasis in China. These molecular data will be helpful to deeply study intra-specific variations within Hymenolepis cestodes in the future.

Epidemiology status of some new emerging Cryptosporidium species and genotypes in humans / 中国人兽共患病学报

Cryptosporidium spp.are protozoan parasites that infect the epithelial cells of the gstrointestinal tract of hosts.In humans,cryptosporidiosis is usually a self-limiting infection in immunocompetent individuals,but severe diarrhea and dissemination to extra-intestinal sites can occur in high-risk individuals,such as the very young,the elderly,immunedeficiency individuals,particularly in HIV-positive patients.So far,molecular epidemiological data have confirmed the presence of 30 species and over 40 genotypes with genus Cryptosporidium,with 21 species and genotypes being found in humans.The majority of human cryptosporidiosis cases are responsible for C.hominis and C.parvum.Human cases caused by C.meleagridis,C.ubiquitum,C.felis and C.canis have been increasing.Besides that,with data accumulation of molecular epidemiology of human cryptosporidiosis,some more Cryptosporidium species and genotypes were newly identified in humans.This paper mainly reviews epidemiology status of these new emerging Cryptosporidium species and genotypes in humans.

The STAT-3 inhibitor WP1066 promotes oral squamous cell carcinoma invasiveness by cisplatin in vitro / 天津医药

Objective To investigate the effect of signal transducers and activators of transcription 3(STAT-3)on sen-sitizing oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miRNA-21. Methods Tscca and Tca8113P160 human tongue squamous cell carcinoma cell lines were employed in this study. WP1066 was used to suppress STAT-3 signaling pathway. Cells were divided into three groups:dimethyl sulphoxide (DMSO) group, cis-dichlorodiamine-platinum (DDP) group and WP1066+DDP group. Transcription level of miR-21 was assessed by real-time PCR, while the expression levels of STAT-3, p-STAT-3, tissue inhibitor of metalloproteinase-3 (TIMP-3) and matrix metalloproteinase-2/9 (MMP-2/9 ) were evaluated by Western blot assay. Matrigel matrix and transwell assay were used to determine cancer cell colony formation and invasive ability respectively. Expression level of miR-21 was examined by luciferase reporter gene as-say. Results Expression levels of STAT-3, pSTAT-3 and miR-21 were significantly suppressed by WP1066 treatment. The diameters of culture colony in cells treated with WP1066 and DDP were smaller than those in control group. The number of tongue cancer cells that migrated through the transwell membrane in WP1066 and DDP treated group was less than that in control group. Additionally, MMP-2/9 expression decreased while TIMP-3 increased dramatically in both cell lines in WP1066+DPP group compared to the other two groups. Conclusion Reduction of STAT-3 can sensitize oral squamous cell carcinoma to cis-dichlorodiamineplatinum via downregulating miR-21. Our study shows that DDP, in combination with WP1066, might be used as a potential target in the treatment of human oral squamous cell cancer.

Signal transducer and activator of transcription-3 inhibitor WP1066 affects human tongue squamous cell carcinoma proliferation and apoptosis in vitro and in vivo / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the antitumour molecular mechanisms of WP1066 (STAT-3 inhibitor ) to human tongue squamous cell carcinoma in vitro and in vivo.</p><p><b>METHODS</b>WP1066 was used to inhibit the p-STAT-3 expression in Tscca human tongue squamous cell carcinoma cell line .Real-time PCR was used to detect the microRNA-21 expression after treatment with DMSO and WP1066. Methyl thiazolyl tatrozolium (MTT) assay was employed to determine cell survival. Flow cytometry (FCM) was used to measure apoptosis. The expression level of STAT-3/p-STAT-3, programmed cell death protein 4 (PDCD-4), antigen 67 (Ki-67), B cell lymphoma 2 (Bcl-2) and cleaved cysteinyl aspartate specific proteinase-3 (CCASP-3) was examined by Western blotting. Luciferase reporter gene assay was conducted to verify the regulation of STAT-3 to microRNA-21. Tscca xenograft tumor model was established in BALB/c nude mice and the tumors were divided into control, DMSO and WP1066 treated groups. The tumor tissues were measured by immunohistochemistry stain and terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay.</p><p><b>RESULTS</b>STAT-3/p-STAT-3 protein was suppressed after treatment with WP1066 (STAT-3:F = 15.336, P = 0.004, p-STAT-3: F = 52.837, P = 0.000). MicroRNA-21 relative expression level was down-regulated (F = 8.197, P = 0.019). Cell survival rate was significantly reduced after treatment with WP1066 than control groups (F = 94.388, P = 0.000). Early apoptosis rate increased after treatment with WP1066 (F = 217.080, P = 0.000) . PDCD-4 and cleaved cysteinyl aspartate specific proteinase-3 (CCASP-3) protein expression was increased after treatment with WP1066 (PDCD-4:F = 8.771, P = 0.017; CCASP-3: F = 26.611, P = 0.001) .Ki-67 and Bcl-2 protein was down regulated (Ki-67:F = 5.854, P = 0.039; Bcl-2:F = 125.502, P = 0.000). Luciferase reporter gene assay proved that STAT-3 combined with specific promoter region of microRNA-21.In vivo, the tumor volume after treatment with WP1066 was significantly smaller than control groups by the end of observation (F = 15.390, P = 0.000) .Immunological histological chemistry indicated that PDCD-4 and CCASP-3 protein expression was up-regulated simultaneously while Ki-67 and Bcl-2 protein of tumor tissue was down-regulated after treatment with WP1066 than control groups. TUNEL assay suggested that apoptosis index rose after treatment with WP1066 than control groups (F = 133.368, P = 0.000) .</p><p><b>CONCLUSIONS</b>WP1066 affected Tscca cancer cell proliferation and apoptosis via inhibiting STAT-3/microRNA-21.WP1066 provided new direction and possibility to human tongue squamous cell carcinoma therapy.</p>

Targeting regulatory T cells and immunotherapy of tumor / 国际肿瘤学杂志

Regulatory T cells (Tregs) possess the function of immune suppression and achieve immunosuppression by the approach of expressing cytotoxic T lymphocyte-associated antigen,perforin-particle enzyme mediated cytotoxic effect,and secretion of cytokines such as IL-10,TGF-β,et al.The antibodies,vaccines and chemical drugs can be applied for targeting FOXP3 + CD25 + CD4 + Tregs,depleting Tregs in peripheral blood and tumor tissues of patients of various forms of cancers.The function of immune suppression can be decreased and antitumor curative effects by immunotherapy can be enhanced.

Molecular identification of Cryptosporidium ryanae isolate from dairy cows in Harbin / 中国人兽共患病学报

One Cryptopsoridium isolate from a 3 month-old calf in a dairy cow farm of Harbin was identified by molecular method, in which the common genes of 18S rRNA and actin used in Cryptosporidium genotyping were selected, the primers were designed, and the approximate 840 bp and 1066 bp fragments of these two genes were amplified respectively by nested PCR. The PCR products were sequenced by two directions, and then the sequences were analyzed by using the soft wares of ClustlX, DNAstar, and PHLIP. The experimental result showed that in the 18SrRNA gene, there was the highest identities compared to other reported isolates of cow-derived C. ryanae (DQ871345, AY587166, and EU410344), with 100% of similarity. Likewise, at actin locus, the present isolate also had 100% similarity with an American cow-derived C. ryanae isolate (EU410345) and a deer-like genotype isolate (AY741309). Phylogenetic analysis indicated that the present isolate shared the same evolution branch in the two gene loci. These analyses demonstrated that the present isolate belonged to C. ryanae. These results provide a significant reference for the study on the species distribution and assessment in the public health significance of Cryptosporidium in cow milk in China.

Study on C21 steroidal glycosides of Cynanchum atratum by LC-MS / 中国中药杂志

High performance liquid chromatography coupled with on-line electrospray tandem mass spectrometry (HPLC/ESI-MS/MS) was used to identify C21 steroidal glycosides in the roots of Cynanchum atratum. The structures of C21 steroidal glycosides were deduced from mass fragments features in positive and negative mode. The constituents of C. atratum were separated and detected. 7 compounds were identified by comparing their ESI-MS/MS data with the reference compounds and 2 compounds were inferred solely by the ESI-MS/MS data. The method is sensitive, and provides good separation and rapid qualitative characterization of C21 steroidal glycosides in the roots of C. atratum.

Effects of Zhitaiqing Granule on aortic endothelial cellsIn rabbit with experimental atherosclerosis / 中华中医药杂志

Objective:To investigate the effects of Zhitaiqing Granule on aortic endothelial cells in rabbit with experimental atherosclerosis. Methods:10 New Zealand rabbits were selected as blank control randomly and fed with normal forage,while the rest rabbits were fed with high-fat forage in formula A. After six weeks,hyperlipidemia models were formed and they were randomly divided into model group,Zhitaiqing treatment group,simvastatin control group and hawthorn tablet control group. From the 7th week,rabbits in treatment and control groups were fed with corresponding drugs while the blank and model groups with the same quantity of normal saline by lavage. After 11 weeks lavage,total cholesterol (CHO),triglyceride (TG),high-density lipoprotein cholesterol (HDL-C),apolipoprotein AI (apo AI),apolipoprotein B (apo B).were detected. With the help of optical microscope,the pathological changes of the aortic arch,thoracic aorta and abdominal aortic intima were observed. Results:In Zhitaiqing group,CHO,LDL-C,and ApoB reduced dramatically (P

Clinical observation of Jianpitongmai soup in treating hyperlipidemia / 中华中医药杂志

Objective:To observe the e ects Of Jianpitongmai soup in regulating blood fat.Methods:120 cases of hyperlipidemia patients were randomly divided into two groups:Jianpitongmai group with 60 cases and simvastatin control group with 60 cases.To compare the treatment e cacy after 8 weeks.Results:Jianpitongmai soup can signi cantly improve indicators of blood fat(P0.05).Conclusions:Jianpitongmai soup can control blood fats comprehensively,and the e ect will be cumulative with the continuation of treatment.