Objective To investigate the correlation of variations of ABCB11 gene and primary intrahepatic stone (PIS).Methods The retrospective case-control study was conducted.The clinical data of 319 patients with PIS and 344 healthy controls who were admitted to the Southwest Hospital of the Third Military Medical University between December 2012 and December 2015 were collected.Three hundred and ninteen patients with PIS and 344 healthy controls were respectively allocated into the PIS and control groups.Twenty-seven exons in ABCB11 gene were detected via polymerase chain reaction (PCR) and DNA sanger sequencing.Observation indicators included:(1) variations and genotype frequency distribution of ABCB11 gene in the 2 groups;(2) correlation analysis between variations of ABCB11 gene and clinical data of patients with PIS:preoperative indicators of liver function test,recurrent episodes of cholangitis,preoperative obstructive jaundice,type and recurrence of stones.Comparison between groups in variations and genotype frequency distribution of ABCB11 gene was analyzed by the Logistic regression.The Kruskal-Wallis H test was performed to explore the correlation between genotype of ABCB11 gene and clinical test results of patients with PIS.Correlation between genotype of ABCB11 gene and clinicopathological data of patients with PIS was analyzed by the chi-square test.Results (1) Variations and genotype frequency distribution of ABCB11 gene in the 2 groups:whole-exome sequencing results showed that synonymous mutations of rs3815675,rs2287616 and rs497692 and missense mutations of rs2287617,rs2287622 and rs118109635 in the PIS group were respectively detected in exon 4,9,24 and 9,13,21.CT genotype frequency of rs118109635 was 4.70％(15/319) in the PIS group and 1.45％ (5/344) in the control group,respectively,with a statistically significant difference [OR=3.49,95％ confidence interval (CI):1.17-10.40,P＜0.05].GG and AG+GG genotype frequency of rs497692 were 46.08％ (147/319),87.46％ (279/319) in the PIS group and 37.79％ (130/344),79.36％ (273/344) in the control group,respectively,with a statistically significant difference (OR=1.73,1.65,95％ CI:1.05-2.83,1.04-2.61,P＜0.05).(2) Correlation analysis between variations of ABCB11 gene and clinical data of patients with PIS:levels of glutamyl-transpeptidase (GGT),alkaline phosphatase (ALP) and direct bilirubin (DBil) in the PIS group were 167 U/L (range,7-1 968 U/L),166 U/L (range,36-1 527 U/L),4 μmol/L(range,1-272 μmol/L) in the CC genotype of rs118109635 and 433 U/L(range,17-864 U/L),232 U/L (range,85-613 U/L),6 μmol/L(range,2-173 μmol/L) in the CT genotype of rs118109635,respectively,with a statistically significant difference (H=6.025,5.879,8.056,P＜0.05).Globulin level of PIS group was respectively 32 g/L (range,20-40 g/L),34 g/L(range,17-50 g/L) and 33 g/L(range,14-49 g/L) in the AA,AG and GG genotype of rs497692,respectively,with a statistically significant difference (H=12.119,P＜0.05).Of 81 patients with recurrence of PIS,GG and GA genotypes of rs2287617 were detected in 78 and 3 patients,respectively,with a statistically significant difference (x2=5.367,P＜0.05);TT,TC and CC genotypes of rs2287622 were detected in 12,39 and 30 patients,respectively,with a statistically significant difference (x2=6.153,P＜0.05).Of 127 patients with obstructive jaundice,116 and 11 patients had CC and CT genotypes of rs118109635,respectively,with a statistically significant difference (x2=7.381,P＜0.05);11,43 and 73 patients had AA,AG and GG genotypes of rs497692,respectively,with a statistically significant difference (x2 =11.364,P＜0.05).Conclusion There is a correlation between rs118109635 and rs497692 of ABCB11 gene and PIS,meanwhile,the above variation loci are associated with obstruction of biliary tract and cholestasis.

AIMTo study the characteristics of neural information encoding of the epileptic networks involved caudate-putamen(CPu) and the hippocampi induced by tetanization of the right CPu in rats.

METHODSExperiments were performed on 59 SD rats. Acute or chronic tetanization of the right CPu (ATRC or CTRC) (60Hz,0.4-0.6 mA, 2 s) was used to induce rat epilepsy.

RESULTS(1) The bursting or primary unit afterdischarges of single neurons were asymmetric in dual hippocampi induced by the ATRC. (2) Continuous sharp waves were observed in ipsilateral or contralateral CPu induced by the CTRC. The oscillatory network seizures with phase shift appeared between two sharp waves in ipsilateral CPu. The frequency of oscillatory waves was negatively correlated with the time and fluctuated from 70 Hz to 110 Hz, then to 35 Hz, and finally to 30 Hz. (3) In the contralateral side primary network after discharges in the CPu were induced by the CTRC. Therefore, the characteristic primary network afterdischarges could be shifted from the CPu or to the HPC, but amplified. On the other hand, HPC sharp waves could be depressed when the CPu network seizures occurred.

CONCLUSIONThe reestablishment of CPu-hippocampal epileptic networks could be transhemispherically promoted by over-activation of the right CPu network, in which bilateral hippocampal neuronal network and CPu neural network were involved in some particular pathophysiological information encoding.

Animals ; Caudate Nucleus ; metabolism ; Epilepsy ; physiopathology ; Hippocampus ; metabolism ; Male ; Neurons ; metabolism ; Putamen ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMThe electrographic and behavioral kindling effects were induced by chronic tetanization of the right caudate-putamen (CPu) to study the target-behavior expression involved in the CPu or hippocampus (HPC) network abnormalities.

METHODSExperiments were performed on 58 SD rats. Tetanization (60Hz,0.4 - 0.6mA, 2s) was delivered into the CPu or the HPC, once a day, for 7-12 days. Animal behaviors were observed every day and depth electrographs were recorded at the beginning or at the end of the experiments.

RESULTSChronic tetanization of the CPu or of the HPC induced: (1) Rhythmic sharp waves in the CPu and paroxysmal epileptiform events in the HPC electrographs. (2) Primary behavioral seizures, secondary behavioral seizures, and kindling effects, including wet dog shakes (WEDS), rearing, face washing, immobility, chewing and head nodding. (3) Lower rate of primary WEDS (P < 0.01), and higher rate of secondary WEDS (P < 0.01) in the CPu-tetanized rats. (4) Longer silent period of behavioral seizures before kindling appeared in the CPu-tetanized rats.

CONCLUSIONKindling effects in the CPu-tetanized rats resembles those in the HPC-tetanized rats. The CPu might participate in the origin of epileptic focus and be involved in reestablishment of limbic epileptic networks, which may be responsible for the target-behavioral seizures.

Animals ; Behavior, Animal ; Caudate Nucleus ; Electric Stimulation ; Epilepsy ; physiopathology ; Kindling, Neurologic ; Male ; Rats ; Rats, Sprague-Dawley ; Seizures ; physiopathology

Objective: To analyze the status of occupational medical examination (OME) institutions in Guangdong Province and the critical control points affecting their future development. Methods: A total of 211 OME institutions registered in Guangdong Province were selected as the research subjects. Their current statue was investigated, and the risk assessment of their future development was carried out based on hazard analysis and critical control point theory. Results: The OME institutions were mainly concentrated in the Pearl River Delta, accounting for 72.0%. The public and private medical and health institutions accounted for 70.6% and 29.4% respectively. The filing rates of different OME categories from high to low in the order were physical factors, chemical factors, dust, other, radiation factors, biological factors (P<0.01), and the filing rates were 89.6%, 88.6%, 84.8%, 63.0%, 7.1% and 1.0%, respectively. The rates of excellence in the field assessment from high to low in the order were hearing atlas analysis personnel, lung function examination operators, chest film reading personnel, chief examination physician, technical director, quality director (P<0.01), and the rates of excellence were 49.4%, 26.2%, 20.6%, 10.8%, 8.2% and 4.6%, respectively. A total of 37 institutions did not submit OME information, of which 23 institutions did not carry out OME work after filing. The required diagnosis rate and confirmed diagnosis rate of suspected occupational diseases in 2020 were 34.3% and 55.3%, respectively. In 2021, the detection rate of suspected occupational diseases was only 1.1‰. Among the institutions with biological monitoring records, 64.1% did not participate in external quality assessment program in laboratory, and the unqualified rate of 38 participating institutions was 55.3%. The risk assessment results show that the future development of OME institutions needs to focus on private institutions, institutions that have not reported OME information, institutions that have missed diagnosis and missed reporting of suspected occupational diseases, institutions that have not participated in external quality assessment program in laboratory and core personnel such as quality managers, technical leaders, and main inspection physicians. Conclusion: OME institutions in Guangdong Province show the characteristics of centralized filing category, centralized filing region and centralized filing institution nature. Combined with the potential risks of OME institutional capacity building, it is necessary to implement classified management, strengthen information construction, supervision and management, implement post responsibilities, and promote the high-quality development of OME institutions.

Objective To analyze the distribution features of newly reported cases of occupational welder′s pneumoconiosis (OWP) in Guangdong Province from 2013 to 2022. Methods Data of newly reported cases of OWP from National Occupational Disease Network Direct Reporting System of Guangdong Province from 2013 to 2022 was analyzed using conventional data analyzing Methods. Results A total of 294 cases of OWP were reported in Guangdong Province from 2013 to 2022 with an overall downward trend. The average age at diagnosis for new OWP cases was 44±6 years old, and the median of dust exposure duration was 11 years. Males accounted for 99.3% of the cases, and 94.2% of the cases were phase Ⅰ OWP. Guangzhou City had the highest regional distribution with 74.8% of the cases. The industry distribution was mainly manufacturing, accounting for 86.7% of the cases. The scale of enterprises was most common in small enterprises, followed by large and medium-sized enterprises, accounting for 38.4%, 30.6% and 24.5% of the cases respectively. The economic types of enterprises were mostly private enterprises, followed by state-owned enterprises, accounting for 54.1% and 32.0% of the cases respectively. Conclusion In Guangdong Province, the newly reported OWP cases were clustered in terms of disease stage, regional distribution, industry, enterprise scale and enterprise economic type. There was a tendency of younger age at diagnosis and shorter dust exposure duration.

OBJECTIVETo investigate the role of microRNA-34a (miR-34a) in regulating the cell cycles of bladder cancer cell line J82 and explore the underlying mechanism.

METHODSJ82 cells were transfected with a miR-34a mimic or an inhibitor to induce miR-34a overexpression or silencing. The RNA level of miR-34a in the transfected cells was detected by real-time PCR, and CD44 expressions at the mRNA and protein levels were detected by real-time PCR and Western blotting. Luciferase reporter assay was used to detect the activation of 3'UTR of CD44, and flow cytometry was performed to analyze the cell cycle changes.

RESULTSThe expression level of miR-34a was significantly increased and CD44 expression significantly lowered in cells transfected with miR-34a mimic; miR-34a inhibitor transfection caused reverse effects on miR-34a and CD44 expressions. MiR-34a mimics downregulated while miR-34a inhibitor enhanced the activation of 3'UTR of CD44 with corresponding changes in the expressions of some cell cycle-related proteins. MiR-34a mimics and miR-34a inhibitor induced opposite changes in J82 cell cycle, which were partly reversed by CD44.

CONCLUSIONMiRNA-34a regulates cell cycles by targeting CD44 in human bladder carcinoma cell line J82.

Cell Cycle ; Cell Line, Tumor ; Down-Regulation ; Humans ; Hyaluronan Receptors ; metabolism ; MicroRNAs ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Transfection ; Urinary Bladder Neoplasms ; pathology

OBJECTIVETo explore the molecular mechanism underlying the biological function of lncRNA PTENP1 in bladder cancer.

METHODSExpressions of PTENP1, PTEN and miR-17 were examined by quantitative reverse transcriptase PCR (qRT-PCR) in 12 bladder cancer tissues. The expression of PTEN was examined by Western blotting in bladder cancer cell lines T24 and 5637 overexpressing PTENP1. Luciferase reporter assay was performed to confirm the targeting of miR-17 to PTENP1 and PTEN. T24 and 5637 cell lines with stable overexpression of PTENP1 and mir-17 were used to investigate effect of PTNE and miR-17 on the function of PTENP1 in bladder cancer.

RESULTSThe expression of miR-17 was up-regulated and PTENP1 and PTEN were down-regulated in bladder cancer tissues, where a positive correlation was found between PTENP1 and PTEN expressions and a negative correlation between PTENP1 and miR-17 (P<0.05). Overexpression of PTENP1 in bladder cancer cell lines T24 and 5637 obviously enhanced the expression of PTEN protein. miR-17 was found to target both PTENP1 and PTEN and promote the growth of bladder cancer. miR-17 could partially restore the tumor-suppressing activity of PTENP1 in bladder cancer.

CONCLUSIONBy binding with miR-17, lncRNA PTENP1 functions as a PTEN competing endogenous RNA (ceRNA) to suppress the progression of bladder cancer.

The purpose of this study was to investigate the impact of leucine-rich repeats and immunoglobulin-like domains 3 (LRIG3) on the biological features of bladder cancer cell lines. The plasmids of over-expressed LRIG3 and the blank plasmid serving as control were transfected into the bladder cancer cell lines, T24, EJ and BIU-87, and the expression levels of LRIG3 mRNA and protein were detected by using real-time PCR and Western blotting. The changes in the cell cycle and apoptosis were examined by using flow cytometry. The invasive ability was measured by Transwell assay, and CCK-8 assays were used to measure the proliferation of cells. As compared with the control group, the LRIG3 mRNA and protein expression levels in LRIG3 cDNA-transfected group were raised significantly (P<0.05). The average number of cells with up-regulated LRIG3 passing through the inserted filter was decreased significantly as compared with the control group (P<0.05). Up-regulation of LRIG3 also could inhibit proliferation and induce apoptosis of T24, EJ and BIU-87 cells. Except BIU-87, the T24 and EJ cells transfected with LIRG3 cDNA were arrested in G(0)/G(1) phase compared to the control group (P<0.05). In conclusion, the over-expression of LRIG3 could influence the cell cycle and invasion, inhibit proliferation and induce apoptosis in the three bladder cancer cell lines.
Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Humans ; Membrane Proteins ; genetics ; metabolism ; Neoplasm Invasiveness ; Up-Regulation ; Urinary Bladder Neoplasms ; metabolism ; pathology

Objective To analyze the surgical treatment and prognostic factors of patients with hepatolithiasis-associated intrahepatic cholangiocarcinoma (HICC).Methods The retrospective case-control study was conducted.The clinicopathological data of 109 patients with HICC who were admitted to the First Affiliated Hospital of Army Medical University between January 2011 and December 2015 were collected,including 40 males and 69 females,aged from 29 to 81 years,with an average age of 55 years.The patients underwent imaging and laboratory examinations before operation,and then the operation plan was worked out according to the results of examination and intraoperative conditions,including radical operation,palliative surgery and simple biopsy.Observation indicators:(1) surgical situations;(2) follow-up;(3) analysis of risk factors affecting postoperative survival of HICC patients.The follow-up using telephone interview and outpatient examination was performed to detect patients' survival up to January 2018.The normality test was done by ShapiroWilk.Measurement data with normal distribution were represented as Mean±SD and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range) and comparison between groups was analyzed using the Wilcoxon rank sum test.Categorical variables were analyzed using the chi-square test.The survival curve and cumulative survival rate were respectively drawn and calculated using the Kaplan-Meier method.The univariate analysis was done using the Log-rank test.The indexes with P＜ 0.05 were incorporated into COX regression model for multivariate analysis in the forward wald.Results (1) Surgical situations:109 patients with HICC underwent surgery.Distribution of hepatolithiasis and HICC in the 109 patients:the stones of 58,26 and 25 patients were located in the left liver,right liver and bilateral sides,respectively.The tumors of 65,31 and 13 patients were located in the left liver,right liver and bilateral sides,respectively.Operation methods:radical operation was performed in 51 cases (combined choledochojejunostomy in 38 cases,regional lymph node dissection in 33 cases,vascular reconstruction in 3 cases and pancreaticoduodenectomy in 2 cases),including 33 with hemihepatectomy and 18 with non-hemihepatectomy.Palliative surgery was performed in 46 cases (local resection of tumors in 13 cases,choledochojejunostomy in 19 cases,gastrointestinal anastomosis in 9 cases,jejunostomy in 13 cases,simple biliary drainage in 5 cases,other operations in 12 cases,some patients combined with a variety of surgical methods),and simple biopsy was performed in 12 cases.Invasion and metastasis in the 109 patients:intrahepatic metastasis occurred in 22 cases,vascular invasion in 52 cases,lymph node metastasis in 55 cases,extrahepatic invasion and metastasis in 58 cases (64 cases were complicated with multiple invasion and metastasis).The operation time and volume of intraoperative blood loss of 109 patients were respectively (359± 170)minutes and (556±382)mL,and 53 patients received intraoperative blood transfusion.Postoperative complications occurred in 35 patients,including pulmonary infection,pleural and peritoneal effusion,abdominal hemorrhage,abdominal infection,bile leakage and organ failure.Twenty-nine of the 35 patients were improved or recovered after treatment,and 6 died (3 died of severe pulmonary infection,1 died of liver failure,1 died of septic shock and 1 died of gastrointestinal hemorrhage).Histopathological characteristics of tumors in the 109 patients:mass type,peritubular infiltration type and intratubular growth type were detected respectively in 50,49 and 10 cases;according to the degree of tumor differentiation,high-differentiated,moderate-differentiated and low-differentiated tumors were detected in 22,50 and 37 cases,respectively.(2) Follow-up:107 patients were followed up for 1-84 months,with a median followup time of 51 months.The median survival time was 25.0 months (range,17.9-32.1 months).The 1-,3-and 5-year overall survival rates were 78.7％,39.4％ and 9.8％,respectively.The 1-,3-and 5-year survival rates were 86.3％,61.8％,20.6％ in patients with radical surgery and 88.4％,19.1％,0 in patients with palliative surgery and 34.6％,0,0 in patients with simple biopsy,respectively,showing a statistically significant difference (x2 =43.237,P＜0.05).(3) Analysis of risk factors affecting postoperative survival of HICC patients:① the results of univariate analysis showed that course of stone,cirrhosis,operation method,tumor diameter,tumor gross classification,degree of tumor differentiation,intrahepatic metastasis,vascular invasion,lymph node metastasis,extrahepatic invasion and metastasis,percentage of neutrophils,total bilirubin (TBiL),direct bilirubin (DBiL),albumin (Alb) and CA19-9 were the related factors affecting the prognosis of HICC patients after surgical treatment (x2 =5.764,8.768,43.273,4.086,11.995,21.910,15.436,6.469,17.181,35.307,24.676,10.691,11.367,5.808,3.907,P＜0.05).② The results of multivariate analysis showed that simple biopsy as operation method,course of stone ≥ 60 months,cirrhosis,low-differentiated tumor,vascular invasion,extrahepatic invasion and metastasis and Alb ＜ 35 g/L were independent factors affecting the prognosis of HICC patients after surgical treatment (hazard ratio =3.317,1.809,1.917,1.882,1.761,2.283,0.502,95％ confidence interval:1.263-8.712,1.132-2.892,1.061-3.463,1.291-2.744,1.087-2.852,1.220-4.271,0.304-0.827,P＜0.05).Conclusions Some patients with chronic hepatolithiasis can progress to HICC,and surgical resection is the first choice.However,due to the high malignancy,there is usually a combination of invasion and mnetastasis,resulting in poor prognosis.The simple biopsy as operation method,course of stone ≥ 60 months,cirrhosis,low-differentiated tumor,perihepatic vascular invasion,extrahepatic invasion and metastasis,Alb ＜ 35 g/L are independent risk factors affecting the prognosis of HICC patients after surgical treatment.

OBJECTIVE: To investigate the mechanism by which long non-coding RNA TUG1 affects bladder cancer cell migration and invasion. METHODS: The expressions of TUG1 and miR-29c-3p were examined by quantitative RT-PCR (qRT-PCR) in 10 bladder cancer tissues and 5 bladder cancer cell lines. Trans-well assay was used to detect the changes in migration and invasion abilities of bladder cancer T24 cells after TUG1 knockdown using RNA interference technique, and the alteration in the expression of CAPN7 was also detected. The expression of CAPN7 was examined in T24 cells overexpressing mir-29c-3p by Western blotting, and luciferase reporter assay was performed to confirm the targeting of miR-29c-3p to TUG1 and CAPN7. The effects of CAPN7 overexpression and sh-TUG1 on the migration and invasion of T24 cells were investigated. RESULTS: The expression of TUG1 was up-regulated and mir-29c-3p was down-regulated significantly in bladder cancer tissue with a negative correlation between their expressions. TUG1 knockdown significantly inhibited the migration and invasion of T24 cells ( < 0.01). Overexpression of mir-29c-3p in T24 cells obviously down-regulated the expression of CAPN7 protein, whose expression was positively correlated with TUG1 expression (=0.4081, =0.0139). The results of luciferase reporter assay confirmed both TUG1 and CAPN7 as the targets of mir-29c-3p. CAPN7 overexpression could partially reverse the tumor suppressing effect of sh-TUG1 in T24 cells. CONCLUSIONS Mir-29c-3p targeting TUG1 affects the migration and invasion of bladder cancer cells by regulating the expression of CAPN7.