This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

To investigate the distribution and biological characteristics of Trueperella pyogenes in caprine mastitis,67 milk samples from were collected from goats with mastitis in 9 dairy goat farms in the Guanzhong area of Shaanxi Province.A total of 21 strains of T.pyogenes were isolated through bacterial separation and culture.PCR amplification was conducted for 8 virulence genes of the 21 isolated strains,and the susceptibility of the strains to 10 antimicrobial drugs was tested u-sing the micro broth dilution method.Nine selected strains were subjected to genomic sequencing using Illumina Nova Seq 6000 next-generation sequencing technology,and their pathogenicity was verified by inoculating mice through the teat canal.The results of virulence gene detection showed that the positive rates of plo,cbpA,nanH,nanP,fimA,fimC-,and fimE genes among the 21 iso-lated strains were 100.0％,38.1％,52.6％,31.6％,61.9％,57.1％,and 52.4％,respectively.In con-trast,the fimG gene was not detected,which was consistent with the results of genomic compari-son by next-generation sequencing.The drug susceptibility test results showed that the susceptibil-ity rates of the isolated strains to clindamycin(CLI),ceftiofur(CEF),and vancomycin(VAN)were 100.0％,while the resistance rates to penicillin(PEN),erythromycin(ERY),sulfamethox-azole-trimethoprim(SXT),florfenicol(FFC),tetracycline(TE),gentamicin(GEN),and enroflox-acin(ENR)were 76.2％,52.1％,52.4％,52.4％,47.6％,42.9％,and 33.3％,respectively.The pro-portion of strains resistant to at least three antimicrobial drugs was 71.4％(15/21).ResFinder a-nalysis of the resistant genes identified 8 types of resistance genes in the genomes of the 9 strains,including ant(2")-Ia and ant(3")-Ia mediating aminoglycoside resistance,cmlA1 and cmx media-ting chloramphenicol resistance,erm(X)and lnu(A)mediating macrolide resistance,sul1 media-ting sulfonamide resistance,and tet(W)mediating tetracycline resistance.Except for tet(W),cm-lA,and erm(X),the other resistance genes were detected for the first time in T.pyogenes in Chi-na.In animal pathogenicity tests,mice inoculated with the isolated strains showed signs of mastitis such as redness,congestion,and inflammatory cell infiltration in the glandular tissue within 24 h,indicating that the isolated strains have pathogenicity to mouse mammary tissue.This study pro-vides preliminary insights into the biological characteristics and pathogenicity of T.pyogenes from caprine mastitis,enriching the data on its drug resistance and genomics,and offering reference and guidance for the prevention and treatment of T.pyogenes infections in large-scale dairy goat farm-ing.

This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

To investigate the distribution and biological characteristics of Trueperella pyogenes in caprine mastitis,67 milk samples from were collected from goats with mastitis in 9 dairy goat farms in the Guanzhong area of Shaanxi Province.A total of 21 strains of T.pyogenes were isolated through bacterial separation and culture.PCR amplification was conducted for 8 virulence genes of the 21 isolated strains,and the susceptibility of the strains to 10 antimicrobial drugs was tested u-sing the micro broth dilution method.Nine selected strains were subjected to genomic sequencing using Illumina Nova Seq 6000 next-generation sequencing technology,and their pathogenicity was verified by inoculating mice through the teat canal.The results of virulence gene detection showed that the positive rates of plo,cbpA,nanH,nanP,fimA,fimC-,and fimE genes among the 21 iso-lated strains were 100.0％,38.1％,52.6％,31.6％,61.9％,57.1％,and 52.4％,respectively.In con-trast,the fimG gene was not detected,which was consistent with the results of genomic compari-son by next-generation sequencing.The drug susceptibility test results showed that the susceptibil-ity rates of the isolated strains to clindamycin(CLI),ceftiofur(CEF),and vancomycin(VAN)were 100.0％,while the resistance rates to penicillin(PEN),erythromycin(ERY),sulfamethox-azole-trimethoprim(SXT),florfenicol(FFC),tetracycline(TE),gentamicin(GEN),and enroflox-acin(ENR)were 76.2％,52.1％,52.4％,52.4％,47.6％,42.9％,and 33.3％,respectively.The pro-portion of strains resistant to at least three antimicrobial drugs was 71.4％(15/21).ResFinder a-nalysis of the resistant genes identified 8 types of resistance genes in the genomes of the 9 strains,including ant(2")-Ia and ant(3")-Ia mediating aminoglycoside resistance,cmlA1 and cmx media-ting chloramphenicol resistance,erm(X)and lnu(A)mediating macrolide resistance,sul1 media-ting sulfonamide resistance,and tet(W)mediating tetracycline resistance.Except for tet(W),cm-lA,and erm(X),the other resistance genes were detected for the first time in T.pyogenes in Chi-na.In animal pathogenicity tests,mice inoculated with the isolated strains showed signs of mastitis such as redness,congestion,and inflammatory cell infiltration in the glandular tissue within 24 h,indicating that the isolated strains have pathogenicity to mouse mammary tissue.This study pro-vides preliminary insights into the biological characteristics and pathogenicity of T.pyogenes from caprine mastitis,enriching the data on its drug resistance and genomics,and offering reference and guidance for the prevention and treatment of T.pyogenes infections in large-scale dairy goat farm-ing.

With bibliometrics method,the 60 published issues from 2011 to 2015 of Chinese Journal of Laboratory Medicine were analyzed from the following aspects:the ratio of fund papers,sources and the number of funds,the collaborating rate of co-authors and the cooperation degree,to reveal the overall level of found paper in medical field.

INTRODUCTIONThe current metastatic category (M) of nasopharyngeal carcinoma (NPC) is a "catch-all" classification, covering a heterogeneous group of tumors ranging from potentially curable to incurable. The aim of this study was to design an M categorization system that could be applied in planning the treatment of NPC with synchronous metastasis.

METHODSA total of 505 NPC patients diagnosed with synchronous metastasis at Sun Yat-sen University Cancer Center between 2000 and 2009 were involved. The associations of clinical variables, metastatic features, and a proposed M categorization system with overall survival (OS) were determined by using Cox regression model.

RESULTSMultivariate analysis showed that Union for International Cancer Control (UICC) N category (N1-3/N0), number of metastatic lesions (multiple/single), liver involvement (yes/no), radiotherapy to primary tumor (yes/no), and cycles of chemotherapy (>4/≤4) were independent prognostic factors for OS. We defined the following subcategories based on liver involvement and the number of metastatic lesions: M1a, single lesion confined to an isolated organ or location except the liver; M1b, single lesion in the liver and/or multiple lesions in any organs or locations except the liver; and M1c, multiple lesions in the liver. Of the 505 cases, 74 (14.7%) were classified as M1a, 296 (58.6%) as M1b, 134 (26.5%) as M1c, and 1 was not specified. The three M1 subcategories showed significant difference in OS [M1b vs. M1a, hazard ratio (HR) = 1.69, 95% confidence interval (CI) = 1.16-2.48, P = 0.007; M1c vs. M1a, HR = 2.64, 95% CI = 1.75-3.98, P < 0.001].

CONCLUSIONSWe developed an M categorization system based on the independent factors related to the prognosis of patients with metastatic NPC. This system may be helpful to further optimize individualized care for NPC patients.

Objective To investigate the efficacy differences of different doses of cyclophosphamide(CTX)among subcategories of diffuse proliferative lupus nephritis(LN). Methods Clinical data of 133 LN patients diagnosed by renal biopsy with class IV or class IV +V who were treated with corticosteroid plus CTX were analyzed retrospectively. The baseline Scr, 24 h urine protein, CTX dosages and prognosis were compared among different dosages for each subcategory. Results The average cumulative dose of CTX within 6 months was(11.1 4.1)g. The high dose group was >12 g, the medium dose group was >6-12 g and the low dose group was ≤6 g. Compared to low dose group, high dose CTX increased the remission rate of class Ⅳ +Ⅴ(67% vs 40%, P=0.314)and chronic renal lesion(43% vs 0%, P=0.212), but such enhancement was not obvious in class Ⅳ(Ⅳ-S: 67% vs 50%, P=0.548, Ⅳ-G: 65% vs 70%, P= 0.560). Difference of overall adverse reactions was not significant between high dose group and low dose group(51% vs 37% ,P=0.224). Conclusion Corticosteroid plus high dose CTX may improve the remission rate of patients with class IV + V and chronic renal lesions.

Objective To investigate the clinicopathologic characteristics, classification and outcome in lupus nephritis(LN)patients with thrombotic microangiopathy(TMA). Methods LN patients with TMA proven by renal biopsy, from January 2000 to February 2009 in our hospital were enrolled. They were classified as poly-immunocomplex deposit group(n =35)and pauci-immunocomplex deposit group(n=25). Their clinicopathologic features and outcome were analyzed retrospectively. Results(l)The incidence of TMA in lupus nephritis was 9.2%(n=62), which presented severe hypertension, prominently elevated serum creatinine, anemia, thrombocytopenia, and was also the poorest prognosis of all the vascular lesion types. The incidence of death/end stage renal disease(ESRD)was 25.0%, with a mortality rate of 13.6%.(2)According to immunocomplex deposit in renal tissue, LN complicated with TMA could be classified as "poly immunocomplex deposit subtype" and "pauci-immunocomplex deposit subtype". The former presented better response to steroid and immunosuppressant therapy, in spite of more active clinicopathologic manifestations. The incidences of death/ESRD of poly subtype and pauci subtype were 8.8% and 32.0% respectively. Conclusions TMA presenting severe manifestations and the poorest prognosis is not rare in LN. LN with TMA may be classified as poly-immunocomplex deposit subtype and pauci-immunocomplex deposit subtype. This classification may be helpful in prognosis because the latter shows bad response to steroid-immunosuppressant therapy.

BACKGROUND: A new type of materials, Collamer, is polymerized with type V collagen and HEMA by Staar company, which is a good material for the establishment of intraocular contact lens(ICL). There is no similar material in our country.OBJECTIVE: To evaluate the biocompatibility of ICL made mainly by collagen-polymer in animals.DESIGN: A randomized controlled trial.SETTING: Testing Center of Shanghai Institute of Biomaterial.MATERIALS: Our study was conducted in the Testing Center of Shanghai Institute of Biomaterial from January 2000 to April 2000. Cell strain: hearty L-929 cells(fibroblast of mouse) after 48-72 hours of passage; Twenty white guinea pig of either gender aged between 1 month and 3 months with a body mass between 300 g and 500 g; Seven healthy adult New Zealand rabbit of either gender(female rabbits were not required pregnant for pyrogenic reaction test) with a body mass between 1.7 kg and 3.0 kg or between 2.5 kg and 3.5 kg(experimental animals were obtained from Shanghai experimental animal center, ordinary grade).INTERVENTIONS: Collagen was used to perform the following biological tests: ① cytotoxicity test (cell growth rate); ② anaphylaxis test; ③ pyrogenic reactions test; ④ subcutaneous implant test. The data were analyzed and evaluated according to criteria.MAIN OUTCOME MEASURES: ① Growth and proliferation of fibroblast in mice; ② Reactions of erythema and edema of every provocation site and every observatory time after the induction of intradermal injection and local patch test; ③ Increase of body temperature of the rabbit after the injection of the extraction of the material into auricle marginal vein; ④ Reactions around the materials after 4 weeks of collagen implantation in rabbits subcutaneously.RESULTS: ① Cytotoxicity test: relative growth rate of collagen group was 99％ to 106%, and the toxicity grade was level 0 - 1. ② Anaphylaxis test:there was no erythema or edema reaction at every provocation site at each time point of collagen group. The skin reaction to collagen-polymer was level 0. ③ Pyrogenic reactions test: the temperature increased in rabbit was below 0.6 ℃ and the total increase of the temperature was below 1.4 ℃ in rabbits. ④ Subcutaneous implant test: there was very little lymphocyte infiltration around the sample in both collagen group and control group with Level I inflammatory reaction. And the evaluation of the formation of fiber cyst was level I.CONCLUSION: Results of material biological evaluation tests demonstrate that this collagen-polymer has high biocompatibility, and can become an ideal material for ICL.