Objective:To explore the function and mechanism of transcription factor En1 in esophageal squamous cell carcinoma (ESCC).Methods:The correlations of En1 with prognosis were analyzed using the overall survival data of 9 397 pan-cancer patients and progression-free survival data of 4 349 pan-cancer patients from The Cancer Genome Atlas (TCGA) database. The En1 expression data in 53 and 155 cases of ESCC and their paired adjacent tissues were from Gene Expression Omnibus (GEO) database and National Genomics Data Center-Genome Sequence Archive(NGDC-GSA)database. Lentivirus was used to generate En1 stable knockout cell lines KYSE180 and KYSE450. The proliferation ability of the cells was detected by cell counting kit 8 and clone formation assay. The migration ability of the cells was detected by Transwell assay. The effect of En1 on the proliferation of ESCC was detected by xenograft experiment in BALB/c-nu/nu mice. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of En1, glioma-associated oncogene family zinc finger 1 (GLI1), glioma-associated oncogene family zinc finger 2 (GLI2) and smoothened (SMO).Results:Pan-cancer data from TCGA showed that patients with low En1 expression had longer overall survival and progression-free survival than patients with high En1 expression ( P< 0.001). Data from GEO and GSA databases also showed a high expression level of En1 in ESCC tissues compared with paired tissues ( P＜0.001). Proliferation was inhibited after knockout of En1 in KYSE180 and KYSE450 cells ( P＜0.001). The colony formation numbers decreased. The colony formation numbers of KYSE180 cells in the shEn1#1 group and the shEn1#2 group were 138.33±23.07 and 127.00±19.70, respectively, significantly lower than that of the shNC group 340.67±12.06 ( P<0.001). The colony formation numbers of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were 65.33±2.52 and 9.00±3.00, respectively, significantly lower than that of the shNC group 139.00±13.00 ( P<0.001). The migration numbers was inhibited after knockout of En1 [the Transwell numbers of KYSE180 cells in the shEn1#1 group and the shEn1#2 group were 66.67±12.66 and 71.33±11.02, respectively, significantly lower than that of the shNC group 334.67±16.56 ( P<0.001). The Transwell numbers of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were 112.33±14.57 and 54.33±5.51, respectively, significantly lower than that of the shNC group 253.33±21.03 ( P<0.001)]. Xenograft model showed a slower growth rate of shEn1#1 and shEn1#2 cell lines ( P＜0.001). The tumor weights of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were (0.046±0.026)g and (0.047±0.025)g, respectively, significantly lower than that of the shNC group (0.130±0.038)g ( P＜0.001). After knockdown of En1, the relative expression levels of GLI1 in KYSE180 cells of the shEn1#1 group and the shEn1#2 group were 0.326±0.162 and 0.322±0.133, and the relative expression levels of GLI1 in KYSE450 cells of the shEn1#1 and shEn1#2 groups were 0.131±0.006 and 0.352±0.050, respectively, which were all lower than that in the shNC group ( P＜0.01). After knockdown of En1, overexpression of GLI1 attenuated the inhibitory effect of knockdown of En1 on cell proliferation ( P＜0.001), colony formation[the colony formation numbers of the shEn1#1-GLI1 group were 151.00±9.54, higher than 102.33±10.02 ( P=0.004) of the shEn1#1-vector group] and migration [the migration numbers of the shEn1#1-GLI1 group were 193.67±10.07, higher than 109.33±11.50 ( P<0.001) in the shEn1#1-vector group]. In clinical samples of ESCC, major regulatory factors of the Hedgehog pathway were up-regulated and the pathway was activated. Conclusion:En1 promotes the proliferation and migration of ESCC cells by regulating the Hedgehog pathway and can be used as a new potential target for targeted therapy of ESCC.

Objective:To explore the function and mechanism of transcription factor En1 in esophageal squamous cell carcinoma (ESCC).Methods:The correlations of En1 with prognosis were analyzed using the overall survival data of 9 397 pan-cancer patients and progression-free survival data of 4 349 pan-cancer patients from The Cancer Genome Atlas (TCGA) database. The En1 expression data in 53 and 155 cases of ESCC and their paired adjacent tissues were from Gene Expression Omnibus (GEO) database and National Genomics Data Center-Genome Sequence Archive(NGDC-GSA)database. Lentivirus was used to generate En1 stable knockout cell lines KYSE180 and KYSE450. The proliferation ability of the cells was detected by cell counting kit 8 and clone formation assay. The migration ability of the cells was detected by Transwell assay. The effect of En1 on the proliferation of ESCC was detected by xenograft experiment in BALB/c-nu/nu mice. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expressions of En1, glioma-associated oncogene family zinc finger 1 (GLI1), glioma-associated oncogene family zinc finger 2 (GLI2) and smoothened (SMO).Results:Pan-cancer data from TCGA showed that patients with low En1 expression had longer overall survival and progression-free survival than patients with high En1 expression ( P< 0.001). Data from GEO and GSA databases also showed a high expression level of En1 in ESCC tissues compared with paired tissues ( P＜0.001). Proliferation was inhibited after knockout of En1 in KYSE180 and KYSE450 cells ( P＜0.001). The colony formation numbers decreased. The colony formation numbers of KYSE180 cells in the shEn1#1 group and the shEn1#2 group were 138.33±23.07 and 127.00±19.70, respectively, significantly lower than that of the shNC group 340.67±12.06 ( P<0.001). The colony formation numbers of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were 65.33±2.52 and 9.00±3.00, respectively, significantly lower than that of the shNC group 139.00±13.00 ( P<0.001). The migration numbers was inhibited after knockout of En1 [the Transwell numbers of KYSE180 cells in the shEn1#1 group and the shEn1#2 group were 66.67±12.66 and 71.33±11.02, respectively, significantly lower than that of the shNC group 334.67±16.56 ( P<0.001). The Transwell numbers of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were 112.33±14.57 and 54.33±5.51, respectively, significantly lower than that of the shNC group 253.33±21.03 ( P<0.001)]. Xenograft model showed a slower growth rate of shEn1#1 and shEn1#2 cell lines ( P＜0.001). The tumor weights of KYSE450 cells in the shEn1#1 group and the shEn1#2 group were (0.046±0.026)g and (0.047±0.025)g, respectively, significantly lower than that of the shNC group (0.130±0.038)g ( P＜0.001). After knockdown of En1, the relative expression levels of GLI1 in KYSE180 cells of the shEn1#1 group and the shEn1#2 group were 0.326±0.162 and 0.322±0.133, and the relative expression levels of GLI1 in KYSE450 cells of the shEn1#1 and shEn1#2 groups were 0.131±0.006 and 0.352±0.050, respectively, which were all lower than that in the shNC group ( P＜0.01). After knockdown of En1, overexpression of GLI1 attenuated the inhibitory effect of knockdown of En1 on cell proliferation ( P＜0.001), colony formation[the colony formation numbers of the shEn1#1-GLI1 group were 151.00±9.54, higher than 102.33±10.02 ( P=0.004) of the shEn1#1-vector group] and migration [the migration numbers of the shEn1#1-GLI1 group were 193.67±10.07, higher than 109.33±11.50 ( P<0.001) in the shEn1#1-vector group]. In clinical samples of ESCC, major regulatory factors of the Hedgehog pathway were up-regulated and the pathway was activated. Conclusion:En1 promotes the proliferation and migration of ESCC cells by regulating the Hedgehog pathway and can be used as a new potential target for targeted therapy of ESCC.

Objective:To investigate the status of occupational stress and its influencing factors among undergraduate interns.Methods:Three hundred and seventeen interns in Subei People's Hospital of Jiangsu Province were collected as research objects. In this study, the Job Content Questionnaire (JCQ), Effort-Reward Imbalance (ERI) Questionnaire and self-compiled general information questionnaire were used to evaluate the status of occupational stress and its influencing factors, and the regression analysis was carried out.Results:The results showed that 101 (31.9%) and 115 (36.3%) undergraduate interns were in significantly higher level occupational stress according to the JCQ and the ERI questionnaire respectively. Logistic regression analysis showed that the risk factors of the included occupational stress included the planning of further studies, more than 40 hours per week for internship, high score of intrinsic engagement and low score of social support. Exercise of more than three times a week was a protective factor for interns' occupational stress.Conclusion:The occupational stress of the undergraduate interns is at a high level and affected by many factors. Schools and hospitals should provide targeted mental health education for interns and improve relevant management policies.

Objective:To investigate the status of occupational stress and its influencing factors among undergraduate interns.Methods:Three hundred and seventeen interns in Subei People's Hospital of Jiangsu Province were collected as research objects. In this study, the Job Content Questionnaire (JCQ), Effort-Reward Imbalance (ERI) Questionnaire and self-compiled general information questionnaire were used to evaluate the status of occupational stress and its influencing factors, and the regression analysis was carried out.Results:The results showed that 101 (31.9%) and 115 (36.3%) undergraduate interns were in significantly higher level occupational stress according to the JCQ and the ERI questionnaire respectively. Logistic regression analysis showed that the risk factors of the included occupational stress included the planning of further studies, more than 40 hours per week for internship, high score of intrinsic engagement and low score of social support. Exercise of more than three times a week was a protective factor for interns' occupational stress.Conclusions:The occupational stress of the undergraduate interns is at a high level and affected by many factors. Schools and hospitals should provide targeted mental health education for interns and improve relevant management policies.

Animals ; Cdc20 Proteins/metabolism* ; Cell Line ; Embryo, Mammalian/embryology* ; Embryonic Development ; Female ; Infertility, Female/metabolism* ; Mice ; Mutation ; Oocytes/metabolism*

To address the increasing need for detecting and validating protein biomarkers in clinical specimens, mass spectrometry (MS)-based targeted proteomic techniques, including the selected reaction monitoring (SRM), parallel reaction monitoring (PRM), and massively parallel data-independent acquisition (DIA), have been developed. For optimal performance, they require the fragment ion spectra of targeted peptides as prior knowledge. In this report, we describe a MS pipe-line and spectral resource to support targeted proteomics studies for human tissue samples. To build the spectral resource, we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on Docker. We then applied the workflow to gen-erate DPHL, a comprehensive DIA pan-human library, from 1096 data-dependent acquisition (DDA) MS raw files for 16 types of cancer samples. This extensive spectral resource was then applied to a proteomic study of 17 prostate cancer (PCa) patients. Thereafter, PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was validated. As a second application, the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma (DLBCL) patients and 18 healthy control subjects. Differentially expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by PRM. These data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker discovery. DPHL is freely accessible at https://www.iprox.org/page/project.html?id=IPX0001400000.

OBJECTIVE： To study the correlation of HLA-DP gene polymorphisms with the immune response to antiviral treatment for hepatitis C virus （HCV） patients. METHODS： A total of 106 HCV Han-nationality patients were collected from our hospital during May 2013 to Aug 2017. All patients received PEG IFNα+ribavirin （RBV） for 48 weeks， and then 24 week follow-up after drug withdrawal. Age， body weight and baseline level of HCV-RNA were recorded. The typing of rs3077 and rs2395309 site of HLA-DP gene were detected by RT-qPCR with Taqman-MGB fluorescent probe. According to treatment outcome， the patients were divided into two groups such as sustained viral response （SVR） group and no-sustained viral response （N-SVR） group. Univariate and multivariate analysis were performed for influential factors （gender， age， body weight index， HCV-RNA baseline level， gene polymorphisms） of immune response to antiviral treatment for HCV patients with Logistic regression model. RESULTS： Among 106 patients， the frequencies of CC， CT and TT genotype at rs3077 site were 40.6％， 35.8％ and 23.6％； those of GG， GA and AA genotype at rs2395309 site were 50.0％， 39.6％， 10.4％， respectively， which were in line with Hardy-Weinberg genetic balance （P＞0.05）. Totally 80 HCV patients were obtained in SVR group， and 26 HCV patients in N-SVR group. The patient’s age， the proportion of CT and TT genotype of rs3077 site and GA and AA genotype of rs2395309 site in SVR group were significantly lower than N-SVR group. The proportion of CC genotype at rs3077 site and GG genotype at rs2395309 site were significantly higher than N-SVR group （P＜0.05）. There was no statistical significance in gender， body weight index or HCV-RNA baseline level between 2 groups （P＞0.05）. Univariate and multivariate analysis showed that gender， body mass index， HCV-RNA baseline level， CC genotype at rs3077 site and GG genotype at rs2395309 site were not related to the immune response to antiviral treatment （P＞0.05）. Age， CT and TT genotype at rs3077 site， GA and AA genotype at rs2395309 site were associated with the immune response to antiviral therapy [OR were 1.135， 1.766， 1.283， 1.218， 1.103， 95％CI were （1.017，1.267）， （1.007，3.100）， （1.038，1.585）， （1.011，1.467）， （1.038，1.172）， respectively， P＜0.05]. CONCLUSIONS： Age and the polymorphisms of HLA-DP gene at rs3077 and rs2395309 site are related to immune response to PEG IFNα+RBV antiviral treatment for HCV Han-nationality patients. Young patients may have higher antiviral immune response rate， while carriers with T and A mutation alleles may have lower antiviral immune response rate.

Objective To observe the effect of platelet rich plasma(PRP)combined with core decompression on the steroid-induced avascular necrosis of the femoral head(SANFH)and on MMP/TIM in rabbits.Methods A total of 42 New Zealand white rabbits were used in this study,and were randomly divided into 3 groups(decompression,combination and control groups,each n=14). The rabbit model of SANFH was established by i.m. injecting prednisolone acetate in the decompression group and combination group. The improved Landesberg method was used to make the platelet rich plasma. The decompression group received core decompression treatment while the combination group received PRP combined with core decompression for bone repair. X-ray photography of the hip joint of the two groups were taken at 2,6 and 10 weeks after the surgery,and the Lane-Sandhu X-ray scores and new bone area ratio were compared. Venous blood samples of the 3 groups were collected and the bilateral femoral heads were taken for further examination. The left femoral heads were used for histopathological observation and the right ones were used to determine the expression of the mRNA of MMP-2,MMP-9, TIMP-1, and TIMP-2. Results The levels of MMP-2 and MMP-9 in the decompression group were higher than that in the combination group and control group after surgery. The levels of TIMP-1 and TIMP-2 in the decompression group were significantly lower than the combination group and control group(P < 0.05). The IL-6 level and the rate of empty bone lacunae in the decompression group were significantly higher than the combination group and control group(P < 0.05), and that of the combination group was higher than the control group(P < 0.05). The combination group had a better joint imaging and histopathological evaluation than the decompression group after surgery. Conclusions Our findings demonstrate that PRP combined with core decompression can exert a positive effect on the MMP/TIMP and bone tissue repair in the treatment of steroid-induced avascular necrosis of the femoral head in rabbits.

Objective To investigate the genetic carrier rate of thalassemia and its gene mutation types as well as the distribution characteristics among couples of reproductive age in Dongfang City of Hainan Province,and to provide a basis for making prevention and control strategies against thalassemia.Methods Samples were collected from 1 000 couples undergoing premarital and pregestational screenings for thalassemia in Dongfang City of Hainan Province from September 2012 to March 2013,in which the positive ones in preliminary screening were further tested by genetic diagnoses and the genotypes were retrospectively analyzed.Results Among 1 000 couples,322 spouses were diagnosed with thalassemia gene mutation and the carrying rate was 16.10％ (322/2 000).In those carriers,246 spouses were α-thalassemia and the carrying rate was 12.30％ (246/2 000),accounting for 76.40％(246/322) of all thalassemia carriers,among them,there were 197 cases of α-deficiency genotype,accounting for 61.18％ (197/322),32 carried mutated α-gene,accounting for 9.94％ (32/322),17 carried both deleted and mutated α-gene,accounting for 5.28％ (17/322);43 spouse were β-thalassemia and the carrying rate was 2.15％(43/2 000);33 spouse were both α-and β-thalassemia and the carrying rate was 1.65％ (33/2 000).In spouses diagnosed with α-thalassemia,the major genotype was-α37/αα,accounting for 19.25％ (62/322);the second ranked was-α4.2/αα,accounting for 17.70％ (57/322),and the third ranked was--SEA/αα,accounting for 8.70％ (28/322).In spouses diagnosed with β-thalassemia,the major genotype was CD41-42/N,accounting for 9.63％ (31/322).Conclusions The population carrying rate of thalassemia in Dongfang City of Hainan Province is high,and its major type is α-thalassemia.For the purpose of decreasing the birth rate of thalassemia,major,local public health department should attach great importance to thalassemia prevention,and strengthen premarital and pregestational screening for thalassemia.

OBJECTIVE: To comprehend the status of knowledge-attitude-practice( KAP) and its effect on blood lead in workers exposed to lead.METHODS: Three hundred and fifty-one first-line lead exposed workers in the smelting industry were chosen as study subjects by cluster sampling method.Blood lead levels in peripheral venous blood were detected.Questionnaire survey was conducted by self-compiled Questionnaire of Knowledge-attitude-practice on Occupational Health in Lead Workers.RESULTS: Among the study subjects,145 workers had abnormal blood lead level( ≥600 μg/L),the abnormal rate was 41.3%.The correct scoring rate in occupational knowledge,attitude,and practice were 25.1%,45.3% and 15.7%,respectively.Multivariate logistic regression analysis results showed that the four risk factors of high blood lead level were wearing no personal protective equipment,not bathing and changing clothes before returning home,not gargling and washing hands before meals,smoking and eating in workplace.CONCLUSION: Poor occupational behaviors can increase the risk of high blood lead level in lead exposed workers.