BACKGROUND: Choriocarcinoma is a kind of trophoblastic neoplasm with highly aggressive phenotypes. Forkhead box D3 (FoxD3) is an embryonic and trophoblastic stem cel transcription factor. It plays important roles in different physical and pathological situations such as embryogenesis, carcinogenesis and tumor progression.
OBJECTIVE:To investigate the role of FoxD3 in choriocarcinoma malignancy and the possible mechanism.
METHODS:The human choriocarcinoma JAR cel line was employed in this study. The mRNA and protein expressions of genes were measured by quantitative RT-PCR (qRT-PCR) and western blot, respectively. The FoxD3 specific short hair RNA was applied to down-regulate gene expression. The cel proliferation was evaluatedin vitro by cel counting assay andin vivo by tumor growth. The migration/invasion was determined by transwel assay. The profile of FoxD3 targeted genes was investigated with an Agilent microarray and verified by qRT-PCR.
RESULTS AND CONCLUSION: The FoxD3 mRNA and protein expressions in JAR cells were significantly higher than those in primarily cultured normal trophoblastic cells. Knockdown of FoxD3 by short hair RNA in JAR cells could inhibit cell proliferation and migration/invasionin vitro, and suppress thetumor growth with decreased β-human chorionic gonadotropin secretionin vivo. A profile of seven focal adhesion molecules (ITGA5, ITGB6, THBS4, COL6A3, VTN, NRXN3 and NLGN1) was verified to be targeted by FoxD3. Furthermore, knockdown of FoxD3 by short hair RNA could decrease the activation of focal adhesion kinase. All these findings suggest the overexpression of FoxD3 can contribute to the aggressive phenotype of choriocarcinoma JAR cells by regulating the profile of focal adhesion molecules and focal adhesion kinase.

Objective By studying the different blood volume blood donors machine adopt platelet product aggregation,platelet content,the machine adopt circulating volume,etc,analysis of different blood volume machine adopt donors on appropriate conditions.Methods The base material of randomly selected 307 blood donors,through blood related index in the study of circulating volume,blood volume,gender,platelets,Hct,Pct count before collection,machine adopt of platelet aggregation and the relationship between the acquisition time;different blood volume,blood platelet count grouping machine adopt the different of platelet aggregation rate,different amount of platelets collect blood volume group,the comparison of blood circulation.The use of statistical software SPSS 17.0,data analysis,analysis including multiple regression analysis,chi-square test,t test,etc.Results 1)The lower count of platelets,Hct,Pct count before collection,the longer the acquisition time,and gender and Pct has nothing to do with the acquisition time the longer the acquisition time,the higher the machine adopt the possibility of platelet aggregation.2)Low blood volume and low platelet count group,machine mining platelet aggregation rate is higher.3)Machine adopt donors of blood volume is higher,the machine adopt the platelet collection amount,the more and the less blood circulation.Conclusion According to the different blood volume blood donors check-up indicators,further carries on the reasonable analysis,optimized machine mining scheme,especially to reasonable arrangement of low blood volume blood donors,so as to improve the quality of platelet collection.

Objective To evaluate the efficacy and safety of therapeutic drug monitoring (TDM ) based vancomycin dose adjustment in patients with gram‐positive infections .Methods A cohort study was designed with 128 inpatients undergoing TDM in Huashan Hospital from January 2005 to September 2014 .The clinical data of these patients were used to analyze the efficacy and safety of vancomycin therapy by Cox model and survival analysis .Results The patients undergoing TDM‐based dose adjustment had a higher daily dose and blood trough concentration ,which may lead to better bacteriological efficacy and overall efficacy .Cox proportional hazards model analysis showed that TDM‐based dose adjustment is a protective factor .No safety‐related risk factor was found .Conclusions TDM‐based vancomycin dose adjustment is important for patients to achieve better outcomes in fighting gram‐positive infections .

Objective To observe the change of JAK-STAT-SOCS signal molecules after interferons alpha acting on the cancerous oral cells in 3 different degrees,namely NOK,DOK and KB cells,and to provide research foundation for the deep understanding of OSCC (oral squamous cell cancer) tumor ceils immune escape mechanism.Methods NOK,DOK,and KB cells were all cultured respectively,and then the third passage cells in the logarithmic growth phase were inoculated in cell culture plate.Blank control group of each hole was added 2 ml complete medium containing 10 ％ FPS.DMSO control group of each hole was added 2 ml complete medium containing 0.1％ DMSO.And in experimental groups containing 10 U/ml,100 U/ml,and 500 U/ml interferons,complete culture medium were added to each hole containing different concentrations of interferons alpha.CP-690550 (100 μmol/L) was added before interferons alpha was added 1 h.All were detected by RT-PCR test and Western blot test after conventional cultured for 24 h.Results RT-PCR detection showed that JAK1 and JAK2 in NOK cells had a small amount of expression,interferons alpha and CP-690550 cells could not influence the expression of JAK1 and JAK2 of NOK group,and there was no statistically significant difference (P ＞ 0.05).Interferons alpha in 100 and 500 U/ml could stimulate the increase of JAK1 and JAK2 expression in DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).CP-690550 could effectively reduce the JAK1 expression of DOK and KB cells,while had no effect on the expression of JAK2,and the differences were statistically significant (P ＜ 0.05).Western blot showed that STAT1,STAT3 and pSTAT3 (Tyr705) all expressed in the control group,while pSTAT1 (Tyr701) didn't express in the control group.Interferons alpha and CP-690550 cells had no effect on STAT1,STAT3 and pSTAT3 (Tyr705) expression of NOK group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increases of pSTAT3 (Tyr705) expression of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).While they had no effect on pSTAT1 (Tyr701) expression.CP-690550 could effectively reduce the pSTAT3 DOK and KB cells (Tyr705) expression,and the differences were statistically significant (P ＜ 0.05).Western blot showed that there were expression of SOCS1 and SOCS3 in control group.Interferons alpha and CP-690550 had no effect on SOCS1 and SOCS3 expression of NOK cell group,and there was no statistically significant difference (P ＞ 0.05).100 U/ml and 500 U/ml of interferons alpha could stimulate the increase of SOCS 1 expression of DOK and KB cells,and differences were statistically significant (P ＜ 0.05).For the expression of SOCS3,no influence.CP-690550 could effectively reduce the expression of SOCS1 of DOK and KB cells,and the differences were statistically significant (P ＜ 0.05).Conclusions Interferons alpha activate DOK JAK1 and KB cells and the expression of JAK2,mainly JAK1 activation.Interferons alpha,by activating DOK JAK1 and KB cells and the expression of JAK2,promote STAT3 phosphorylation in Tyr705 locus.Interferons alpha,by promoting STAT3 phosphorylation,further promote the expression of SOCS1,which plays the role in inhibiting interferons alpha and reducing the apoptosis.

Objective To observe the transdifferentiation of renal tubular epithelial phenotype in allograft biopsy samples of patients with various rejections,and to analyze the association between rejection and transdifferentiation.Method Immunohistochemistry (SP method) was applied to detect α-SMA expression in tubular epithelial cells from 55 renal allograft biopsy samples with various rejection.Results Positive α-SMA expression was found in all the atrophic tubular epithelial cells adjacent to cytoplasm of basement membrane,which indicated the atrophic renal tubular epithelial cells appeared the phenotypic transdifferentiation.Positive α-SMA was also detected in some renal epithelial cells without atrophy.No phenotypic change was found in 7 cases without obviously rejection.Among 28 cases of acute T-cell-mediated rejection IA grade,α-SMA positive expression rate of non-atrophy renal epithelial cells was 25％-50％ in 1 case and 10％-25％ in 3 cases.Among 14 cases of more severe acute rejection group IB grade,α-SMA positive expression rate was over 50％ in 1 case,25％-50％ in 2 cases and 10％-25％ in 2 cases.Conclusion When acute T-cell-mediated rejection becomes more serious in renal allograft,the phenotype transdifferentiation aggravates in renal tubular epithelial cells.

Objective: To explore the effect of the aortic arch type on technical indicators in patients with carotid artery stent implantation.
Methods: We retrospectively analyzed 224 consecutive patients treated in Fu Wai hospital for unilateral carotid artery stent implantation from 2011-01 to 2012-12. We summarized the catheter category, type and the operating techniques including ① retracement, turn and insertion of the catheter, ② retracement, turn of catheter+the guidance of guide wire,③ retracement, turn of catheter+the guidance of guide wire+the supporting of another catheter, ④ using special graphic catheter+the guidance of guide wire+the supporting of another catheter. The procedural X-ray exposure time, dosage of contrast agent and operation related complications were recorded. According to Myla classiifcation, the aortic arches were divided into Myla I, Myla II and Myla III types.
Results: There were 7/224 (3.1%) patients with Myla I aortic arch, 113 (50.4%) with Myla II aortic arch and 104 (46.4%) with Myla III aortic arch. A total of 48/104 (46.2%) Myla III patients used special techniques (tech③, tech④), it was more than the patients with Myla I, (1/7,14.3%) and Myla II (17/113, 15.0%), P<0.01. The patients with Myla III aortic arch had the longer X-ray exposure time and used the higher dose of contrast agent, all P<0.01. The procedural success rate in patients with
Myla III was 96.2%, it was lower than those with Myla I (100%) and Myla II (100%), P=0.045. The procedural complication rate in patients with Myla III was 22.1%, it was higher than those with Myla I (0%) and Myla II (8.9%), P=0.007.
Conclusion: The aortic arch type is the important inlfuential factor for the techniques used in carotid stent implantation. There were more dififculties and complications for stent implantation in patients with Myla III aortic arch.

Objective: To investigate the duration of Norovirus (NoV) shedding among infected school children during a NoV outbreak in a kindergarten,and to provide scientitic basis for epidemic prevention and control. Methods: Specimens and epidemiological data were collected from suspected cases, and specimens were detected using real-time RT-PCR to determine whether or not infecting with NoV. Specimens were collected every 3-7 days from NoV-infected children until specimens became negative for NoV. Results: A total of 14 suspected cases were reported, and 12 of them were infected with NoV. The average duration of NoV shedding was (26.58±17.94)d. The specimens among 9 from 12 Nov-infected cases were positive at 7 days, 8 NoV-intected cased remained positive at 14 days and 7 Non-infected cased at least 21 days. Conclusion Since NoV shedding duration among NoV-infected children tends to longer than their isolation time during outbreaks, reinforcement of hygiene practices among these school children is especially necessary to reduce the risk of virus secondary transmissions after their return to school.

Using mutation PCR, we cloned the target gene containing 421-480nt (141-160aa) and 598-639nt (200-213aa) of VP1 gene of foot and mouth disease virus (FMDV) into the deleted region (508-532aa) of Nsp2 gene of a highly pathogenic porcine reproductive and respiratory syndrome virus derived vaccine strain (HuN4-F112) that was used as vector. The recombinant cDNA was in vitro transcribed followed by transfection of BHK-21 cells for 36 h. Then, the supernatant of the cell culture was continuously seeded to monolayer of MARC-145 cells for recovery of the recombinant virus. CPE was obviously visible after a couple of passages in the seeded MARC-145, and the rescued virus (designated as rPRRSV-F112-O/VP1ep) was identified by Mlu I digestion, sequencing and immunofluorescence assay. Meanwhile, expression of inserted FMDV epitopes was also detected by indirect immunofluorescence assay with polyclonal antibodies against VP1 protein of FMDV. The analysis of biological characteristics shows that the titer of the rescued recombinant PRRSV (TCID50 = -log10(-6.75)/0.1 mL) was similar to its direct parental virus rHuN4-F112-delta508-532, but higher than rHuN4-F112.
Animals ; Antigens, Viral ; immunology ; Base Sequence ; Capsid Proteins ; immunology ; Cell Line ; Cysteine Endopeptidases ; genetics ; Epitopes ; genetics ; Foot-and-Mouth Disease ; immunology ; prevention & control ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Molecular Sequence Data ; Mutation ; Porcine respiratory and reproductive syndrome virus ; genetics ; immunology ; Recombination, Genetic ; Swine ; Transfection ; Vaccines, Attenuated ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology ; Viral Vaccines ; genetics ; immunology

OBJECTIVETo analyze the diagnostic value of larval excretory-secretory antigen in Angiostrongylus cantonensis (LESA) infection.

METHODSA.cantonensis larvae harvested from mice brain were cultured in vitro. The LESA and the adult worm antigens of A.cantonensis (AWA) were collected and analyzed using SDS-PAGE and Western blotting. Two ELISA systems were established using the two antigens (LESA-ELISA and AWA-ELISA) to detect the serum spectra from different sources.

RESULTSSDS-PAGE and Western blotting displayed fewer protein and antigen bands for LESA than for the adult antigen. Two distinct bands of LESA (with relative molecular masses of 40 000 and 26 000) showed reactivity with the sera from patients with A. cantonensis infection. The serum levels of IgG and IgM antibodies to LESA increased at the beginning of infection in mice, reaching the peak on day 5 after infection and decreased on day 10. Compared with AWA-ELISA, LESA-ELISA showed a lower seropositive ratio in suspected patients with A.cantonensis, with also a lower cross-positive ratio in patients with schistosomiasis and clonorchis sinensis.

CONCLUSIONLESA possesses fewer antigen reaction bands than AWA. Although with a slightly lower positive ratio than AWA, LESA has a higher specificity for detecting serum antibodies in suspected cases of A.cantonensis infection, and therefore shows a potential for the diagnosis of angiostrongyliasis especially in the early stage and in current infection.

Angiostrongylus cantonensis ; immunology ; Animals ; Antigens, Helminth ; immunology ; Enzyme-Linked Immunosorbent Assay ; Humans ; Larva ; immunology ; Mice ; Mice, Inbred BALB C ; Rats ; Rats, Sprague-Dawley ; Strongylida Infections ; diagnosis ; parasitology

Objective:To explore the relationship between CDKN1B expression and clinicopathological features in colorectal cancer.Methods:Human Protein Atlas (HPA) database and Gene Expression Profiling Interactive Analysis (GEPIA) database were used to analyze the expression of CDKN1B in colorectal cancer tissues and its relationship with the prognosis of colorectal cancer. The data of 98 patients with colorectal cancer who underwent surgery from January 2020 to December 2021 at Yixing Clinical College of Yangzhou University Medical School were retrospectively analyzed, and pathological specimens were collected. Immunohistochemistry method was used to detect CDKN1B protein expression level in colorectal cancer and paracancerous normal tissues (2 cm from the tumor site) and the correlation of CDNK1B expression with clinicopathological characteristics was analyzed.Results:The results of bioinfomatics analysis and the prediction from HPA database and GEPIA database suggested that the expression level of CDKN1B in colorectal cancer was lower than that in the normal colorectal tissues; In HPA database, the 5-year overall survival rate of patients in the CDKN1B high expression (425 cases) was higher than that of those in the CDKN1B low expression (172 cases) (65% vs.51%), and the difference in the overall survival of both group was statistically significant ( P < 0.001). GEPIA database staging module analysis showed that CDKN1B gene expression level was correlated with the pathological stage of patients with colorectal cancer ( P = 0.033). Immunohistochemistry analysis showed that CDKN1B expression was localized in the nucleus and cytoplasm. The proportion of patients with CDKN1B high expression in colorectal cancer tissues was lower than that in paracancerous normal tissues [18.37% (18/98) vs. 90.82% (89/98), P < 0.01]. The proportion of CDKN1B high expression in cancer tissues of colorectal cancer patients with poor differentiation [poor differentiation vs. high-middle differentiation: 3.70% (1/27) vs. 23.94% (17/71)], lymph node metastasis [metastasis vs. non-metastasis: 6.38% (3/53) vs. 29.41% (15/45)], TNM higher stage [stage Ⅳ vs. Ⅲ vs. Ⅱ vs. Ⅰ: 5.00% (1/20) vs. 13.95% (3/33) vs. 20.59% (8/30) vs. 36.36% (6/15)] was lower (all P < 0.05), while there were no statistically significant differences in the proportion of patients with CDKMB high expression in colorectal cancer tissues among different subgroups stratified by gender, age and tumor size (all P > 0.05). Conclusions:CDKN1B is mainly expressed in the nucleus and cytoplasm, and is lowly expressed in colorectal cancer. The lower CDKN1B expression may indicate the poorer prognosis of patients. CDKN1B can be used as a marker for clinical diagnosis, treatment and prognosis evaluation of colorectal cancer.