[Auxiliary diagnosis and prognosis evaluation of KIF4A, RAD51AP1 and CDKN3 in esophageal cancer].

To investigate the expression of mRNA in esophageal cancer (ESCA) tissues and its potential and diagnostic and prognostic value by high-throughput sequencing data. Using the Cancer Genome Atlas Program (TCGA) database in USA by integrative bioinformatics analysis methods, the gene expression profiles and clinical data of 173 patients with ECSA were collected. The mRNA expression levels in ESCA tissue and para-cancerous tissue samples were analyzed using DESeq2, edgeR and limma to screen the differentially expressed genes (DEGs). DEGs-related protein network diagrams were drawn. GO and KEGG function enrichment analysis were performed and the hub genes were screened and the survival analysis of hub genes was analyzed. Genes related to the prognosis of ESCA were selected and their prognostic value in ESCA was analyzed. Finally, the receiver operating characteristic curve was drawn to evaluate its diagnostic value. The results showed that using TCGA cancer data, a total of 620 up-regulated DEGs and 668 down-regulated DEGs with significant differential expression between ESCA and para-cancerous tissues were screened. DEGs were mainly involved in receptor complexes, ubiquitin ligase complexes, etc., playing GTPase activity, phospholipid binding, and other molecular functions, and participating in the regulation of intracellular substance transport, small molecule metabolism, and other biological processes. Protein functional enrichment analysis showed that these proteins were mainly enriched in the IL-17 signaling pathway, TNF signaling pathway, Toll-like receptor signaling pathway, Epstein-Barr virus infection, neutrophil extracellular trap formation, and other pathways involved in the formation and development process of ESCA. Survival analysis showed that the overall survival rate of ESCA patients with high expression of KIF4A, RAD51AP1, and CDKN3 was significantly shortened, and the difference was statistically significant (P<0.05). Furthermore, the areas under the curve (AUC) of KIF4A, RAD51AP1, and CDKN3 for diagnosing esophageal cancer were 0.956, 0.951 and 0.979, respectively, with sensitivities and specificities both exceeding 80%. Additionally, ROC results of the combined diagnostic model of these three genes showed an AUC of 0.979, with sensitivities and specificities of 0.914 and 1, respectively. This indicates that KIF4A, RAD51AP1 and CDKN3 have individual or combined auxiliary diagnostic value for ESCA. In conclusion, KIF4A, RAD51AP1 and CDKN3 have high diagnostic efficiency for ESCA, and their increased expression is closely related to the prognosis, suggesting that these three genes could be used as auxiliary diagnostic and prognostic factors for ESCA.

Genome-wide association studies uncover genes associated with litter traits in the pig.

Litter traits are critical economic variables in the pig industry as they represent a production indicator that can serve to determine sow fertility. In this study, a genome-wide association study on litter traits, including total number born (TNB), number born alive (NBA), litter birth weight (LBW), average birth weight (ABW), and piglet uniformity (PU), was carried out on two pig breeds (Yorkshire and Landrace). A total of 3 637 pigs of both breeds were genotyped using the GeneSeek GGP Porcine 50K SNP BeadChip. A mixed linear model (MLM) and fixed and random model circulating probability unification (FarmCPU) were employed in the genome-wide association studies for litter traits using combined data from the two pig breeds and data from each breed separately. Additionally, the heritability of traits was estimated using three methods-pedigree-based best linear unbiased prediction (PBLUP), genomic best linear unbiased prediction (GBLUP), and single-step best linear unbiased prediction (ssGBLUP)-and was found to lie between 0.065 and 0.1289, 0.0478 and 0.0938, 0.0793 and 0.0935, 0.1862 and 0.2163, and 0.0327 and 0.0419 for TNB, NBA, LBW, ABW, and PU, respectively. We also compared the genomic prediction accuracies and unbiasedness for litter traits of the three BLUP models. Our results indicated that the ssGBLUP method provided higher predictive accuracies and more rational unbiasedness compared with the PBLUP and GBLUP methodologies. Furthermore, based on their possible roles, eight candidate genes (INHBA, LEPR, HDHD2, CTNND2, RNF216, HMX1, PAPPA2, and NTN1) were identified as being linked with litter traits. In the middle of the test, these genes were found to be connected with pig metabolism and ovulation rate. Our results provide the insights into the genetic architecture of litter traits in pigs, and the potential single nucleotide polymorphisms (SNPs) and candidate genes identified may benefit economic profits in pig-breeding industry and contribute to improve litter traits.

[Investigation on the health status of workers exposed to benzene in one province].

Objective: To explore the health status of occupational benzene exposure workers, and to provide a scientific basis for the development of reasonable health monitoring and effective protective measures. Methods: In March 2019, the occupational health surveillance data were collected including blood pressure, electrocardiogram, blood routine, urine routine, liver function, etc of 7810 benzene contact workers in 150 enterprises in Jiangxi Province in 2017, to analyze and assess their health status. Results: Among the 7810 benzene workers, there were 5451 males and 2359 females; the average age was (40.5±9.9) years; and the median benzene working age was 3.5 years. The detection rate of hypertension was 17.0% (734/4317) , the abnormal rate of urine routine was 15.7% (1227/7810) , the abnormal rate of liver function was 8.6% (356/4147) , and the abnormal rate of electrocardiogram was 12.3%(963/7810). The detection rates of low count number of leukocytes, platelets, neutrophils and occupational contraindications were 4.6%(360/7810) , 1.4%(108/7810) , 4.2%(330/7810) and 1.4%(110/7810) , respectively. The detection rates of low count number of leukocytes, platelets and neutrophils in female were all higher than those in male (P<0.05). The detection rates of low count number of platelets, neutropenia and occupational contraindications increased with age and working age (P<0.05 ). There were significant differences in the detection rates of low count number of leukocytes, platelets, neutrophils and occupational contraindications among benzene workers in different economic types (P<0.05) , and the highest among foreign companies, followed by private enterprise. There were statistically significant differences in the detection rates of low count number of platelets, neutrophils and occupational contraindications in benzene workers of different enterprise sizes (P<0.05) , and the highest was found in micro enterprises, followed by small enterprises. Conclusion: In 2017, the occupational health status of workers exposed to benzene in Jiangxi province is not optimistic. It is necessary to strengthen the occupational health supervision of small and micro-sized enterprises, foreign enterprises and private enterprises, take the initiative to improve health surveillance, and effectively protect the physical and mental health of workers.

[Association between soluble receptor for advanced glycation end products (sRAGE) and coronary and cerebral atherosclerosis].

Objective: To investigate the Association between soluble receptor for advanced glycation end products (sRAGE) and coronary and cerebral atherosclerosis. Methods: A total of 232 consecutive patients who synchronously undertook coronary angiography and craniocerebral CT angiography (or total cerebral angiography) were included between May 2018 and December 2018 in Beijing Tiantan Hospital, Capital Medical University in this study. Patients were divided into the control group (without coronary artery disease (CAD) and cerebrovascular stenosis (CVS), n=55), CAD group (n=118), CVS group (n=11), concomitant CAD and CVS group (CAD+CVS, n=48). Plasma sRAGE level was measured by enzyme-linked immunosorbent assay (ELISA) and compared among the four groups. The relationship between sRAGE and Gensini Score (GS) and cerebrovascular stenosis severity was assayed. sRAGE levels were compared among low, middle and high GS group as well as between extracranial and intracranial arteries stenosis. Results: The levels of sRAGE in CAD group (1.96 µg/L) were higher than those in the control group (1.66 µg/L, P=0.025) or the CVS group (1.53 µg/L, P=0.013). However, no significant difference in sRAGE level was found between the groups of CAD and CAD+CVS (1.89 µg/L, P>0.05). Meanwhile, sRAGE was positively associated with GS in the entire study population (r=0.153, P=0.023) or in the diabetic patients (r=0.242, P=0.017). The sRAGE leves in both middle GS and high GS groups were higher than those in low GS group (P<0.05). No association between sRAGE and CVS severity and vascular count. Additionally, no significant difference in levels of sRAGE was found between extracranial (1.84 µg/L) and intracranial arteries stenosis (1.66 µg/L, P=0.523). Conclusion: Plasma sRAGE level is positively associated withseverity of CAD, but its association with cerebral atherosclerosis needs further studies.

[Efficacy and outcome of transcatheter closure of patent foramen ovale in patients with cryptogenic stroke].

Objective: To investigate the efficacy and outcome of transcatheter patent foramen ovale (PFO) closure in patients with cryptogenic stroke (CS). Methods: Sixty consecutive patients with cryptogenic stroke who undertook transcatheter PFO closure between May 2015 and September 2017 in Beijing Tiantan Hospital were enrolled in this prospective study.Transcranial Doppler (TCD) bubble test was performed and right-left shunt(RLS) was confirmed in all patients.Closure success rate,effective closure rate, complications, recurrence of ischemic stroke and new onset atrial fibrillation were evaluated. Results: A total of 60 patients (42 male,age range 24-68 (47±11)years) were included in the study.PFO size (motionless state) was (1.6±0.6)mm.RLS before closure was graded and 11 patients had moderate RLS and 48 patients had large RLS (include 41 patients who experienced shower or curtain effect).Closure success rate was 100% (60/60).No severe complications were observed.At 6 months,45 patients completed TCD bubble test.Of these, 4 patients suffered from moderate to large residual and thus effective closure rate was 91%(41/45).The mean follow-up period was 2-29 (median 12) months. During the follow-up, only 1 patient experienced recurrent cerebral infarction.New onset atrial fibrillation was not detected. Conclusion: Transcatheter PFO closure is effective,safe and related with a good outcome in reduction of recurrent CS for patients with PFO.

[Molecular epidemiologic study on Mycobacterium tuberculosis from drug resistance monitoring sites of Guangdong Province, 2015].

Objective: To understand the characteristics of Mycobacterium tuberculosis (MTB) in epidemiology and distribution from Guangdong Province, and to explore the risk factors associated with drug resistance. Methods: A total of 225 clinical strains of MTB collected from 5 drug resistance monitoring sites of Guangdong Province in 2015 were tested by Regions of Difference 105 (RD105) deletion test and 15 loci mycobacterial interspersed repetitive units (MIRU) were used for genotyping. Gene clustering was analyzed using BioNumerics7.6. Drug susceptibility test was tested by proportion method. The statistical analysis used chi-square test and multivariate logistic regression. Results: There were 158 (70.2%) Beijing family strains from the 225 cases. Hunter-gaston index of MIRU loci varied from each other. The MTBs from Guangdong Province were categorized into 2 gene clusters by clustering analysis in which the rate of cluster of complexⅠwas significantly higher than complexⅡ(χ(2) values were 9.331, P values were 0.020). It was found by multivariate logistic regression that Qub11b was associated with resistance to rifampicin and isoniazid (P values were 0.013, 0.012 respectively.), ETR F with resistance to isoniazid, streptomycin, ethambutol and ofloxacin (P values were 0.039, 0.040, 0.023 and 0.003 respectively), Mtub21 with resistance to capreomycin (P values were 0.040), and QUB26 with resistance to ethionamide (P values were 0.047). Conclusions: The genes of MTB from Guangdong Province were of polymorphisms and the distribution of strains were stable. QUB11b, ETR F, Mtub21 and QUB26 could be related to biomarkers for predicting drug resistance.

[Artemisinin inhibits proliferation of gallbladder cancer cell lines through triggering cell cycle arrest and apoptosis].

OBJECTIVE: To evaluate the effects of artemisinin on proliferation, cell cycle and apoptosis of gallbladder cancer cells. METHODS: Gallbladder carcinoma cell lines(GBC-SD and NOZ)were cultured in vitro. The effects of artemisinin in different concentration on proliferation of the two cell lines in vitro were examined using MTT assay. The cell cycle distribution of GBC-SD and NOZ cells 24 h after treatments with artemisinin(20 µmol/L) were examined using flow cytometry. The apoptosis of GBC-SD and NOZ cells 24 h after treatments with artemisinin (20 µmol/L) were examined using Annexin V/PI staining.The expressions of p-ERK1/2, CDK4, cyclin D1, p16, cytochrome C and caspase-3 were examined by Western blot assay. t-test and one way ANOVA were used to evaluate the differences between two groups and more than two groups, respectively. RESULTS: The cell proliferation was significantly inhibited by artemisinin, the IC50 of artemisinin against GBC-SD and NOZ cells were 14.05 µmol/L and 12.42 µmol/L, respectively.Artemisinin induced cycle arrest, and G1 population of GBC-SD and NOZ cells increased to 74.60% and 78.86%. Cell apoptosis and apoptotic population of GBC-SD and NOZ cells were increased to 15.67% and 16.51% after dealt with artemisinin, respectively. In addition, expression of p16 was increased, and expressions of p-ERK1/2, CDK4 and cyclin D1 were down-regulated by artemisinin(all P<0.05). Cytochrome C was released from mitochondria to cytoplasm leading to the activation of caspase-3 and PARP after dealt with artemisinin(P<0.05). CONCLUSION: The inhibition effect of artemisinin on the proliferation gallbladder cancer cells is accompanied by down-regulation of ERK1/2 signaling pathway, G1 phase arrest and triggering caspase-3-mediate apoptosis.

Expression of Smac induced by the Egr1 promoter enhances the radiosensitivity of breast cancer cells.

Breast cancer is one of the most prevalent cancers worldwide. Moreover, despite advances in antineoplastic therapies, induction of tumor cell death without off-target cytotoxicity remains a challenge. However, recent developments in localized radiotherapy and gene therapy have provided an opportunity to explore the potential for these strategies to be additive for the induction of cell death in tumor cells. Here, a novel adenoviral shuttle vector containing the proapoptotic gene Smac under the control of the ionizing radiation (IR)-induced Egr1 promoter was constructed. Following the transient transfection of the construct into MCF-7 and MDA-MB-435 breast cancer cell lines, acute and abundant expression of Smac was observed in response to IR treatment. Further analysis confirmed that the induction of Smac expression resulted in a decrease in cell viability, a slower rate of cell growth, a higher level of apoptosis and altered cell cycle progression. Using a clonogenic assay, IR-induced Smac expression was also found to significantly sensitize Smac-expressing cells to radiation-induced cell death. Taken together, these data suggest that Smac expression driven by the Egr1 promoter has the potential to serve as a radiotherapy-dependent gene therapy agent.

Polymorphisms of the angiotensin II type 1 receptor gene affect antihypertensive response to angiotensin receptor blockers in hypertensive Chinese.

The renin-angiotensin-aldosterone system plays a key role in regulating blood pressure by maintaining vascular tone and the water/sodium balance. Many antihypertensive drugs target the renin-angiotensin-aldosterone system, but the effect differs considerably among hypertensive patients. We investigated whether genetic variants of the angiotensin II type 1 receptor are associated with blood pressure response to angiotensin II receptor blockers in hypertensive Chinese patients. After a 2-week single-blind placebo run-in period, 148 patients with mild-to-moderate primary hypertension received monotherapy with 80 mg/day telmisartan and then were followed up for 8 weeks. The 1166A/C, 573T/C, -810A/T, and -521C/T polymorphisms of the AT1R gene were determined through PCR and RFLP analysis. The relationship between these polymorphisms and changes in blood pressure was observed and evaluated after 8 weeks of treatment. Patients with the AT1R -521CC genotype had a significant reduction in diastolic blood pressure compared to those carrying the T allele. No significant reduction in blood pressure was found in individuals with the 1166A/C, 573T/C, or -810A/T polymorphisms of the AT1R gene. We conclude that only the AT1R -521CC genotype is associated with a significant decrease in blood pressure in response to telmisartan treatment in Chinese hypertensive patients.

Radiosensitization of breast cancer cells by TRAIL-endostatin-targeting gene therapy.

One of the key issues in cancer radiotherapy research is to sensitize tumor cells to the cell killing effects of ionizing radiation while leaving normal tissues intact. One potential approach to achieve this is gene-radiotherapy, i.e. a combination of radiation therapy and gene therapy. It is to choose certain exogenous radiation-inducible regulatory genes, for example, early growth response-1 (Egr-1), and transcript its downstream tumor-therapeutic genes under ionizing radiation so as to kill the tumor cells synergistically by the expressed gene products together after transfection and irradiation exposure. In this study, we engineered a plasmid encoding both TRAIL and endostatin under the control of the radiation-inducible Egr-1 promoter, and evaluated its anti-tumor efficacy in combination with radiotherapy. Our plasmid showed significant efficacy in up-regulating the levels of TRAIL and endostatin proteins after transfected into breast cancer cells and exposed to X-ray irradiation. The detected cellular effects in vitro manifested that TRAIL-endostatin-based gene therapy could enhance radiosensitizing effects in breast cancer cells in terms of tumor cell growth inhibition, promoting apoptosis and the induction of cell cycle arrest. In summary, our results suggest that TRAIL-endostain-targeting approach might be a promising method to sensitize solid tumors to radiation therapy.

The fluorescence recovery of polydiacetylene/fluorophore vesicles by interaction with cetyltrimethylammonium bromide.

Polydiacetylene (PDA) has been widely used as a biosensor candidate because it exhibits specific blue-red transition upon varied external stimulations, such as heat, pH, or mechanical pressure. We prepared mixed vesicles using the 10,12-pentacosadiynoic acid (PCDA) and fluorescence probe BO558, and investigated the interaction of surfactant with PCDA/BO558 vesicles. Experiments exhibited that the fluorescence of BO558 was significantly quenched owing to the ene-yne conjugated backbones in the PCDA vesicles. However, the fluorescence was gradually recovered in the presence of cetyltrimethylammonium bromide (CTAB) to the system. Meanwhile, the turbidity of the mixed solution decreased greatly with the increase of CTAB concentration. We investigated the mechanism of the fluorescence quenching and recovery in the system of PCDA/BO558 vesicles and CTAB by UV-vis spectrum, stable fluorescence, transmission electron microscopy. The morphology of vesicles transformed with the change of surfactant concentration.

An effective dose of valdecoxib in experimental mouse models of pain.

The effects of selective cyclooxygenase-2 (COX-2) inhibitors in biological functions are frequently investigated in animal models. However, there is little data on their analgesic efficacy in experimental animals. This study aimed to determine whether oral gavage of 5 mg/kg valdecoxib in mice is active as an analgesic at this dose and whether it is associated with therapeutic blood levels. A nonselective COX inhibitor, ketorolac, was also investigated for comparison. A total of 106 C57 BL/6N mice were administered a single oral dose of 5 mg/kg of valdecoxib, ketorolac or placebo. The antinociceptive effects of both drugs were tested using hot-plate and formalin tests. For the hot-plate test, reaction time (latency) of the mouse before jumping was recorded. The total time that the mouse spent on licking/biting the injected paw (with dilute formalin) was recorded in the formalin test. Apart from the behavioral tests, plasma concentrations of the drugs at this dose were also determined. Mice were fed with 5 mg/kg of either valdecoxib or ketorolac. Blood samples were collected between 1 and 9 h postingestion. Valdecoxib and ketorolac concentration in the plasma was determined by high performance liquid chromatography with ultraviolet detection (HPLC-UV). Effective antinociception was observed for both drugs in the hot-plate test from 75 min to 2 h after oral dosing. Also, both drug treatments showed a significantly reduced nociceptive response in the second phase in the formalin test (20-30 min after injection). Both valdecoxib and ketorolac showed plasma concentrations comparable to the therapeutic concentrations in humans. A single oral dose of valdecoxib or ketorolac (5 mg/kg) is able to produce a therapeutic analgesic effect in mice.

Expression of HSP70-2 gene during germ cell apoptosis in rat unilateral cryptorchid testes.

To investigate the role of Hsp70-2 gene in germ cell apoptosis induced by heat stress, its expression changes were examined in rat normal and unilateral cryptorchid testes by using in situ hybridization, immunohistochemistry, and northern blot analysis techniques. The results showed that the expression level of Hsp70-2 gene declined slightly at the early stage of germ cell apoptosis, and dropped dramatically when most of the germ cells were undergoing apoptosis on day 7.5 after the induction of cryptorchidism. This report suggests for the first time that Hsp70-2 gene might not inhibit the apoptosis of germ cells at the early stage in cryptorchid testes. Hsp70-2 gene does not belong to the immediate early related genes that are responsible for germ cell apoptosis induced by heat stress.

[Germ cell apoptosis and regulation in testis].

Both spontaneous and induced germ cell apoptosis occur during the process of germ cell differentiation in testis, which is an important mechanism to remove surplus and abnormal germ cells. It has been known that germ cell apoptosis is under the control of endocrine, cell social association and gene. To elucidate the molecular mechanisms by which germ cell apoptosis is regulated and the elements of the death machinery will provide a basis for effective management of male infertility as well as more targeted approaches to male contraception.

Cloning of novel temperature-related expressed sequence tags in rat testis during spermatogenesis.

Spermatogenesis needs the relatively cool environment of the scrotum in most mammals, it would be arrested when the testis was exposed to abdominal temperature. In this study, we have used a differential display PCR technique (DD-PCR) to screen temperature-related ESTs during spermatogenesis (TRS) in scrotal testes through a unilateral cryptorchid rat model after in situ analysis of testis cell DNA fragmentation. We reported here the cloning and sequencing of three such ESTs: TRS1, TRS3, and TRS4. Northern blot analysis confirmed that they were expressed specifically in scrotal testes. In situ hybridization showed that TRS1 was mainly expressed in the spermatocytes and the round spermatids in scrotal testis. Homology searches revealed that TRS1 and TRS3 were unknown cDNA sequences, and TRS4 was identical to a known EST whose function had not been reported. TRS1, TRS2, and TRS3 were first found to be temperature-related during spermatogenesis.

Localization of plasminogen activator and inhibitor, LH and androgen receptors and inhibin subunits in monkey epididymis.

Epididymis is a site of sperm maturation and storage. Limited and directed-proteolysis regulated by plasminogen activator (PA), plasminogen activator inhibitor type-1 (PAI-1) and other related factors may play an essential role in these processes. Our previous studies have demonstrated that rat epididymis expressed luteinizing hormone receptor (LHR), tissue type (t) and urokinase type (u)PA, mRNAs, and tPA activity was stimulated in vitro by human chorionic gonoadotrophin (HCG). In the present study we further examined localization of mRNAs for tPA, uPA, LHR, androgen receptor (AR), as well as inhibin subunits alpha, betaA and betaB in rhesus monkey epididymis. Using in-situ hybridization with digoxygenin-labelled cRNA probes, we have demonstrated that tPA and PAI-1 mRNAs were localized in epithelial cells of adult monkey epididymis. uPA mRNA was localized in the same areas, but to a much smaller extent. tPA, uPA and PAI-1 mRNAs were greatly expressed in the caput and corpus of adult epididymis than in other regions. In-vitro experiments showed that both tPA and uPA activities in epididymal cells were dramatically stimulated by HCG, but not by follicle stimulating hormone (FSH). LHR (but not FSH receptor) and AR mRNAs were localized in the epithelial cells of the epididymis. However, LHR mRNA was detected in both adult and immature infant monkeys, whereas AR was found only in the adult. Inhibin alpha, betaA and betaB mRNAs were also detected in this organ, betaA mRNA being more strongly expressed in the caput than in other regions of the epididymis. We suggest that LH and androgen may be the key hormones in coordination with the PA-PAI-1 system in regulating epididymal differentiation and sperm maturation.