A brand new era of cancer immunotherapy: breakthroughs and challenges / 中华医学杂志(英文版)

Immunotherapy has opened a new era in cancer treatment. Drugs represented by immune checkpoint inhibitors have led to important breakthroughs in the treatment of various solid tumors, greatly improving the survival rate of cancer patients. Many types of immunotherapeutic drugs have become widely available; however, their efficacy is variable, and relatively few patients with advanced cancer experience life-altering durable survival, reflecting the complex and highly regulated nature of the immune system. The research field of cancer immunotherapy (CIT) still faces many challenges in pursuing the broader social goal of "curing cancer." Increasing attention has been paid to strengthening the understanding of the molecular or cellular drivers of resistance to immunotherapy, actively exploring more effective therapeutic targets, and developing combination therapy strategies. Here, we review the key challenges that have emerged in the era of CIT and the possible solutions or development directions to overcome these difficulties, providing relevant references for basic research and the development of modified clinical treatment regimens.

Correlation of circulating tumor DNA EGFR mutation levels with clinical outcomes in patients with advanced lung adenocarcinoma / 中华医学杂志(英文版)

BACKGROUND@#Circulating tumor DNA (ctDNA) is a promising biomarker for non-invasive epidermal growth factor receptor mutations (EGFRm) detection in lung cancer patients, but existing methods have limitations in sensitivity and availability. In this study, we used the ΔCt value (mutant cycle threshold [Ct] value-internal control Ct value) generated during the polymerase chain reaction (PCR) assay to convert super-amplification-refractory mutation system (superARMS) from a qualitative method to a semi-quantitative method named reformed-superARMS (R-superARMS), and evaluated its performance in detecting EGFRm in plasma ctDNA in patients with advanced lung adenocarcinoma.@*METHODS@#A total of 41 pairs of tissues and plasma samples were obtained from lung adenocarcinoma patients who had known EGFRm in tumor tissue and were previously untreated. EGFRm in ctDNA was identified by using superARMS. Through making use of ΔCt value generated during the detection process of superARMS, we indirectly transform this qualitative detection method into a semi-quantitative PCR detection method, named R-superARMS. Both qualitative and quantitative analyses of the data were performed. Kaplan-Meier analysis was performed to estimate the progression-free survival (PFS) and overall survival (OS). Fisher exact test was used for categorical variables.@*RESULTS@#The concordance rate of EGFRm in tumor tissues and matched plasma samples was 68.3% (28/41). At baseline, EGFRm-positive patients were divided into two groups according to the cut-off ΔCt value of EGFRm set at 8.11. A significant difference in the median OS (mOS) between the two groups was observed (EGFRm ΔCt ≤8.11 vs. >8.11: not reached vs. 11.0 months; log-rank P = 0.024). Patients were divided into mutation clearance (MC) group and mutation incomplete clearance (MIC) group according to whether the ΔCt value of EGFRm test turned negative after 1 month of treatment. We found that there was also a significant difference in mOS (not reached vs. 10.4 months; log-rank P = 0.021) between MC group and MIC group. Although there was no significant difference in PFS between the two groups, the two curves were separated and the PFS of MC group tended to be higher than the MIC group (not reached vs. 27.5 months; log-rank P = 0.088). Furthermore, EGFRm-positive patients were divided into two groups according to the cut-off of the changes in ΔCt value of EGFRm after 1 month of treatment, which was set at 4.89. A significant difference in the mOS between the two groups was observed (change value of ΔCt >4.89 vs. ≤4.89: not reached vs. 11.0 months; log-rank P = 0.014).@*CONCLUSIONS@#Detecting EGFRm in ctDNA using R-superARMS can identify patients who are more likely sensitive to targeted therapy, reflect the molecular load of patients, and predict the therapeutic efficacy and clinical outcomes of patients.

Value of serum gamma-glutamyl transpeptidase combined with direct bilirubin in the diagnosis of biliary atresia in infants / 中国当代儿科杂志

OBJECTIVE@#To study the value of serum gamma-glutamyl transpeptidase (GGT) combined with direct bilirubin (DB) in the diagnosis of biliary atresia.@*METHODS@#A total of 667 infants with cholestasis who were hospitalized and treated from July 2010 to December 2018 were enrolled as subjects. According to the results of intraoperative cholangiography and follow-up, they were divided into biliary atresia group with 234 infants and cholestasis group with 433 infants. The two groups were compared in terms of age of onset, sex, and serum levels of total bilirubin (TB), DB, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bile acid (TBA), and GGT. A receiver operating characteristic (ROC) curve analysis was performed for indices with statistical significance, and the area under the ROC curve (AUC) and the optimal cut-off value for diagnosis were calculated.@*RESULTS@#The biliary atresia group had a significantly younger age of onset than the cholestasis group (P0.05), while the biliary atresia group had significantly higher serum levels of TB, DB, TBA, and GGT than the cholestasis group (P<0.05). GGT combined with DB had the highest AUC of 0.892 (95% confidence interval: 0.868-0.916) in the diagnosis of biliary atresia. At the optimal cut-off values of 324.0 U/L for GGT and 115.1 μmmol/L for DB, GGT combined with DB had a sensitivity of 79.8% and a specificity of 83.2% in the diagnosis of biliary atresia.@*CONCLUSIONS@#GGT combined with DB has high sensitivity and specificity in the diagnosis of biliary atresia and can be used as an effective indicator for diagnosis of biliary atresia in infants.

Anorectal malignant melanoma:diagnosis, treatment, and prognostic analysis of 36 cases / 中国肿瘤临床

Objective:To analyze the clinicopathological features, diagnosis, and treatment of anorectal malignant melanoma (ARMM), and to explore its prognostic factors and misdiagnosis. Methods:A total of 36 patients with ARMM were enrolled in this study from January 2000 to November 2016 in Nanfang Hospital, Zhujiang Hospital, and Guangdong Provincial Hospital of Traditional Chinese Medicine. Results: The clinical manifestations of ARMM were not specific. The odds of misdiagnosis were as high as 52.8% in this study. The 1-and 3-year survival rates were 75%and 35%, respectively, with median survival time of 24.51 months. Survival rate was correlated with tumor size, invasion depth, clinical stage, and lymph node metastasis (P<0.05), but was not related to patient age and gender. The median survival time of the three groups of patients (surgery alone, surgery-based combination therapy, untreated) were 39.21, 26, and 15 months. The difference was not statistically significant. No difference in survival was found between patients under-going abdominoperineal resection and wide local excision. Conclusion:ARMM has poor prognosis and is easily misdiagnosed as a ma-lignant tumor. The prognostic factors are tumor size, invasion depth, clinical stage, and lymph node metastasis. Surgical treatment can extend survival. To avoid misdiagnosis and prolong survival, early diagnosis and early treatment are recommended.

Claudin-3 expression in colorectal carcinoma and its significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of claudin-3 in colorectal carcinoma and its association with the occurrence, progression and prognosis of colorectal cancer.</p><p><b>METHODS</b>Forty surgical specimens of colorectal carcinoma and 22 adjacent normal tissues resected between October, 2010 and January, 2013 at Nanfang Hospital were examined for claudin-3 expression using immunohistochemistry, which was analyzed in association with the clinicopathological parameters and the survival of the patients.</p><p><b>RESULTS</b>Claudin-3 was expressed mainly on the cell membrane, and its positivity rate was significantly higher in cancer tissues than in normal tissues (92.50% vs 59.09%, P<0.05). In 13 cases claudin-3 expression was detected in both the cancer tissues and adjacent normal tissues with average expression scores of 4.538 and 3.269, respectively (P<0.05). In the cancer tissues, the strongly positive expression rate was significantly higher in poorly differentiated tissues (85.71%) than in well (21.43%) and moderately (36.48%) differentiated tissues (P<0.05), and was higher in cases with lymph node metastasis than in those without (61.11% vs 22.72%, P<0.05). The strongly positive expression rate of claudin-3 was not correlated with the patients'age, gender, tumor location or tumor size (P>0.05). Of the 33 cancer patients followed up, 14 had a postoperative survival time no longer than 3 years and 19 had longer survival time, and their average expression scores differed significantly (4.50 vs 3.526, P<0.05).</p><p><b>CONCLUSION</b>Claudin-3 is over-expressed in colorectal cancer tissues, and its high expression may promote the occurrence and progression of colorectal cancer. Claudin-3 may serve as a molecular biomarker for early diagnosis and prognostic evaluation.</p>

Study on the Molecular Docking of Penehyclidine Optical Isomers and Muscarinic Receptor Subtypes / 中国药房

OBJECTIVE:To study the affinity of penehyclidine optical isomers to muscarinic(M)receptor subtypes,and pro-vide reference for revealing the action targets and efficacy selectivity of penehyclidine. METHODS:Homology modeling,molecu-lar docking and other molecular simulation technologies were used to analyze and predict the binding energy of 4 optical isomers to M receptor subtypes and judge its affinity by comparing the binding energy of different optical isomers R1 (3R,2′R),R2 (3R, 2′S),S1(3S,2′R),S2(3S,2′S)with M receptor subtypes M1-M5. RESULTS:All the 4 optical isomers can dock into the ac-tive sites of M receptor subtypes,and different optical isomers showed great differences in the molecular docking with different M receptor subtypes. Penehyclidine isomers showed larger binding energy to M3,the binding energy of 4 optical isomers ranged in 5736.519-5907.143 kcal/mol. The binding energy of R1 to M1 was 1190.041 kcal/mol;while those of other optical isomers to each receptor subtype were lower or negative. CONCLUSIONS:R1 shows the affinity to M1 receptor. And all the 4 optical isomer show the affinity to M3.

Application of ultrasound-guided modified-fascia iliaca compartment block in the supine position in elderly patients with hip fracture / 临床麻醉学杂志

Objective To compare the analgesic effect of the ultrasound-guided modified-fascia iliaca compartment block with ultrasound-guided fascia iliaca compartment block injection in the treat-ment of elderly patients with hip fracture.Methods Sixty elderly patients with hip fracture,17 males and 43 females,falling into ASA physical status Ⅱ or Ⅲ,were randomly divide into two groups (n=30 each):ultrasound-guided modified fascial iliaca compartment block group (group M)and ultra-sound-guided fascial iliaca compartment block group (group F).The patients in group M received M-FICB using ultrasound-guided injection of 0.4% ropivacaine 5 ml in obturator nerve,15 ml in the fas-cial iliac space.The patients in group F received ultrasound-guided injection of 0.4% ropivacaine 20 ml in the fascial iliac space.FICB or MFICB was performed 20 min before epidural anesthesia in group F or group M respectively.The time of ultrasound-guided nerve block was recorded,and the onset time of femoral nerve,lateral femoral cutaneous nerve and obturator nerve block were recorded in the two groups.Visual analogue pain scores (VAS)were recorded before nerve block (T0 ),after nerve block,10 min (T1 ),20 min (T2 ),placing spinal anesthesia position (T3 ),and postoperative 24 h (T4 ).Results The onset time of obturator nerve block in group M was significantly shorter than that in group F [(4.1±1.4)min vs (10.1 ±3.9)min,P <0.05].The time of ultrasound-guided nerve block has no difference between the two groups [(2.2 ± 0.5 )min vs (2.1 ± 0.5 )min].Compared with group F,the VAS scoress at T1-T3 were lower in group M (P <0.05).Compared with T0 ,the VAS scores at T1-T4 decreased in both groups (P < 0.05 ).Conclusion Ultrasound-guided fascia iliaca compartment block is more effective in reducing the VAS scores during the supine position and reducing postoperative pain.

Therapeutic value of endoscopical therapy for small gastric stromal tumor / 中华消化内镜杂志

Objective To investigate the safety and effectiveness of endoscopical therapy for small grastric stromal tumor( d<2 cm) . Methods Clinical records of 80 patients dignosed as having small grastric stromal tumor by pathology and immunohistochemistry were retrospectively analyzed. Endoscopic or laparo?scopic or open surgery was performed, so patients were divided into endoscopic treatment group ( n=37) and surgical treatment group ( n=43) accordingly. Perioperative period data, effectiveness and follow?up data were analysed. Results Thirty?five cases( 94?6%) of endoscopic treatment group were successfully treated by endoscopic surgery. Intraoperative perforation occurred in 8 cases, among which 2 cases transferred for conventional operations.Pneumoperitoneum occurred in 2 cases (better after symptomatic treatment), while no serious infection or perioperative death occurred. There were no significant differences in the complete re?section rate[ 91?89% ( 34/37) VS 100?0%( 43/43) , P=0?095) , the incidence of postoperative complica?tions[5?71%(2/35)VS 2?33%(1/43),P=0?855)or the improvement of postoperative symptoms[93?10%( 27/29) VS 85?71% ( 30/35) , P=0?589] ,while statistically significant differences were found in operation time[(37?41±13?45)min VS(84?56±38?37)min, P=0?000], intraoperative blood loss[(5?65±5?88)ml VS(31?48 ±39?57)ml, P=0?000], postoperative feeding time[(2?47±0?61)d VS(3?26 ±1?27)d, P=0?001], postoperative hospital stay[(5?76 ±2?28)d VS(7?64±2?99)d,P=0?022],and medical expenses [(18 554?4±9 736?45)yuan VS(31 138?11±1 206?24)yuan, P=0?000]. Conclusion Endoscopic re?section is safe and effective for small grastric stromal tumor with minimal invasion,simple procedure, faster recovery and less economic burden for the patients.

Clinical and pathologic features of small gastrointestinal stromal tumor / 中华消化内镜杂志

Objective To investigate the clinical and pathologic features of small gastrointestinal stromal tumors (small GISTs,d ＜ 2.0 cm).Methods Medical records of 95 patients undergoing surgery (endoscopic surgery,thoracoscopic/laparoscropic surgery and open surgery)and diagnosed as having GISTs by pathology and immunohistochemistry in Nanfang hospital from October 2003 to June 2014 were retrospectively analyzed.Based on clinical and pathological results,correlation analyses between risk factors for endoscopic ultrasonography(EUS) and Mitotic count(MI),clinicopathologic parameter and NIH risk classification were performed.Results Among 95 cases (104 lesions),88 were single,while 7 were multiple;81.7％ (85/104) small GISTs arose from stomach,including 87.1％ (74/85)in middle-upper stomach;5 cases (5.3％) presented calcification of different degrees,3 cases(3.2％) presented local necrosis and 2 cases (2.1％) with arrangement of epithelioid cells;88 cases (92.6％) were very low grade of NIH risk classification,6 cases (6.3％) were intermediate risk and 1 case(1.1％) was high risk.Positive rates of CD34 and CD117 were 95.8％ (91/95) and 96.8％ (92/95) respectively.The risk factors (border,mucosal surface,echo and heterogeneity) of EUS had no correlation with mitotic count(P＞0.05).The correlation analysis between clinicopathologic features and NIH risk classification revealed tumors more than 1.5 cm had a striking correlation with NIH risk classification (P＜ 0.05).Conclusion Most small GISTs,single or multiple,located at middle-upper stomach,were of very low or low risk,and have a favorable prognosis.But it has worse biological behavior and a higher grade risk when the diameter is more than 1.5 cm,intervention should be recommended.

Brain gray matter abnormalities revealed by voxel-based morphometry in patients with chronic low back pain / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the morphometric abnormalities of brain gray matter (GM) in patients with chronic low back pain (CLBP).</p><p><b>METHODS</b>Thirty patients with CLBP and 30 healthy individuals were enrolled and examined with a 3.0 T magnetic resonance (MR) scanner. High-resolution T1 structural MR data were acquired and data analysis was performed using voxel-based morphometry (VBM) in FMRIB Software Library. The morphological differences were compared between the two groups.</p><p><b>RESULTS</b>s Compared with the healthy control subjects, patients with CLBP showed decreased GM volumes in several brain cortical areas including the bilateral superior frontal gyrus, right frontal pole, left insular cortex, left middle and left inferior temporal gyrus (P<0.05, after TFCE correction). Increased GM volumes were found in the patients in the subcortical structures including the left thalamus, bilateral putamen, bilateral nucleus accumben and right caudate nucleus (P<0.05, after TFCE correction).</p><p><b>CONCLUSION</b>Patients with CLBP have different patterns of GM abnormalities in different brain regions, characterized by reduced GM volume in cerebral cortical regions and increased GM volume in the subcortical nuclei. Such changes might be associated with the maladaptation of the brain in chronic pain state.</p>

Interaction between folate deficiency and aberrant expression related to fragile histidine triad gene in the progression of cervical cancerization / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the interaction between folate deficiency and aberrant expression related to fragile histidine triad (FHIT) gene in the progression of cervical cancerization.</p><p><b>METHODS</b>A total number of 80 patients with histological diagnosis of cervix inflammation (CI), 55 cervical intraepithelial neoplasm I (CIN I), 55 cervical intraepithelial neoplasm II/III (CIN II/III) and 64 cervical squamous cell carcinoma (SCC) were included in this study. Levels of serum folate were detected by microbiological assay method and the methylation status of FHIT gene CpG islands was tested by methylation-specific PCR (MSP). FHIT protein levels were measured by Western blot. In vitro, cervical cancer cell lines CaSki (HPV16-positive) was treated with different concentrations of folate. Proliferation and apoptosis of cells, methylation of FHIT gene and the levels of FHIT protein expression were measured in each group. All analyses were performed with SPSS (version 17.0) statistical software. Differences among groups were assessed by chi-square test, Kruskal-Wallis test. Spearman correlation, and the interaction effects were evaluated by additive model.</p><p><b>RESULTS</b>The levels of serum folate (H = 59.08, P < 0.001) and FHIT protein expression (H = 50.93, P < 0.001) decreased gradually along with the severity of cervix lesions, while the methylation rates of FHIT gene CpG islands increased (trend χ² = 28.34, P < 0.001). Both levels of serum folate levels and FHIT protein expression were positively correlated (r = 0.213, P = 0.001), with an additive interaction seen between them in CIN I, CIN II/III, SCC groups. In vitro, both rates related to proliferation inhibition (r = 0.98, P < 0.001) and apoptosis (r = 0.99, P < 0.001) together with the levels of FHIT protein expression (r = 0.97, P < 0.001) were all increased gradually with the increase of folate concentration while the methylation status of FHIT gene CpG islands all changed from positive to negative gradually.</p><p><b>CONCLUSION</b>Results from our study revealed that both folate deficiency and FHIT protein aberrant low expression might increase the risk of developing cervical cancer and cervix precancerous lesions, and thus play a synergistic action in the progression of cervical cancerization.</p>

Comparison of endoscopic variceal ligation and pericardial devascularization in treatment of portal hypertension / 临床肝胆病杂志

Objective To compare the clinical efficacy of endoscopic variceal ligation versus pericardial devascularization in the treatment of portal hypertension.Methods The clinical data of 101 cirrhotic patients with gastroesophageal varices and variceal hemorrhage from January 2010 to January 2012 were analyzed.Fifty-three patients received endoscopic variceal ligation,and forty-eight patients received pericardial devascularization.Postoperative changes in liver function and hypersplenism were compared between the two groups.The rate of rehaemorrhagia and incidence of postoperative complications after surgery were compared as well.Continuous data were expressed as mean ±SD,and categori-cal data were expressed as number of cases or percentage.Comparison of continuous data between the two groups was made by independent-samples t test,and comparison of categorical data was made by chi-square test.Results After surgery,the variceal ligation group showed no significant changes in liver reserve function,while the albumin level was significantly decreased in the pericardial devascularization group (t=2.512,P0.05),but significant increases in the counts of white blood cells and platelets were detected in the pericardial devascular-ization group (P0.05), 6 months,7.5%vs 8.3%(χ2 =0.021,P>0.05),and 1 year,9.4%vs 8.3%(χ2 =0.038,P>0.05).The incidence rates of postopera-tive complications in the two groups were 24.5%and 50%,respectively (χ2 =7.040,P<0.05).Conclusion Compared with pericardial devascularization,endoscopic variceal ligation causes fewer microlesions,preserves liver function,and leads to a lower incidence of postopera-tive complications.However,if hypersplenism is observed in the cirrhotic patients with gastroesophageal varices and variceal hemorrhage,peri-cardial devascularization can be used to control the hypersplenism and prevent esophageal hemorrhage.

Relationship between TGF-β1 Gene Polymorphism and Liver Cirrhosis after Hepatitis B Virus Infection:A Meta-Analysis in Chinese Population / 天津医药

Objective To evaluate the relationship between genetic polymorphism of transforming growth factor (TGF)-β1 and susceptibility of liver cirrhosis after hepatitis B virus infection in Chinese population. Methods CBM, VIP, CNKI, Wanfang technological periodical full-text databases and Pubmed from set up to July, 2013 were electronically searched to identify case-control studies on the relationship between genetic polymorphism of TGF-β1 promoter 509 site, co-don 869 site and liver cirrhosis after hepatitis B virus infection. The data were quantitatively analyzed by RevMan 5.1 soft-ware after assessing the quality of included studies. Results Six case-control studies were selected for Meta-analysis based on our inclusion and exclusion standards. The results of Meta-analysis showed that the pooled OR value for liver cir-rhosis among Chinese patients after hepatitis B virus infection with T allele of TGF-β1 gene at promoter 509 was 1.02 (95%CI:0.67-1.54), the pooled OR values for patients with TT and CT genotypes were 0.80 (95%CI:0.36-1.78). OR values for pa-tients with C allele of TGF-β1 gene at codon 869 was 1.05 (95%CI:0.69-1.62), the pooled OR values for patients with CC and CT genotypes were 0.98 (95%CI:0.48-2.00). No significant publication bias was found. Conclusion The genetic poly-morphism of TGF-β1 at promoter 509 and codon 869 showed no association with susceptibility of liver cirrhosis after hepati-tis B virus infection in Chinese population.

Influence of folate on fragile histidine triad gene expression, cell proliferation and apoptosis in cervical cancer cell lines / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation, expression of mRNA and protein of fragile histidine triad (FHIT)gene in cervical cancer cells.</p><p><b>METHODS</b>Cervical cancer cell lines including CaSki (HPV16-positive) and C33A (HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR (MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR, respectively.</p><p><b>RESULTS</b>Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.98, P < 0.001) and the apoptosis rate increased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.99, P < 0.001) along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1 µg/ml and 10 µg/ml, partially positive at 100 µg/ml and 250 µg/ml, but negative at 500 µg/ml and 1 000 µg/ml in both C33A and CaSki cells. Comparing with the control group, the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells, and showing that the FHIT gene mRNA expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.94, P < 0.001) and protein expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.97, P < 0.001) both increased along with the increase of folate concentration.</p><p><b>CONCLUSION</b>Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro, so would reverse the aberration mRNA and protein expression of FHIT gene.</p>

Influence of folate on fragile histidine triad gene expression,cell proliferation and apoptosis in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.

Influence of folate on fragile histidine triad gene expression,cell proliferation and apoptosis in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.

Effect of DNA methyltransferase 1 and methyl-CpG-binding protein 2 abnormal expression on cervical lesions and related interaction / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the effect of DNA methyltransferase 1 (DNMT1) and methyl-CpG-binding protein 2 (MeCP2) on cervical cancer and cervix precancerous lesion.</p><p><b>METHODS</b>74 patients with cervix squamous cell carcinoma(SCC), 52 patients with cervical intraepithelial neoplasm I (CIN I), 60 patients with cervical intraepithelial neoplasm II - II (CIN II-III)and 58 patients with histologically diagnosed cervix inflammation(CI), were included in this study. Information as demography, reproductive history, life style, HPV infection were collected. Western Blot were used to detect the expression of DNMT1 protein and MeCP2 protein. Real-time PCR was used to detect the expression of DNMT1 and MeCP2 mRNA.</p><p><b>RESULTS</b>Levels of DNMT1 and MeCP2 protein expression increased gradually with the deterioration of cervical lesion (H = 94.33, P < 0.001;F = 21.580, P < 0.001). Along with the deterioration of cervical lesion, levels of DNMT1 and MeCP2 mRNA expression were gradually increasing( F = 4.758, P = 0.003; F = 7.804, P < 0.001). Data from Correlation analysis showed that both protein (r = 0.287, P < 0.001) and mRNA(r = 0.179, P = 0.005)were positive correlated with DNMT1 and MeCP2.</p><p><b>RESULTS</b>of our study indicated that there was an additive interaction between high-expression of DNMT1 protein and high-expression of MeCP2 protein in SCC or CIN II-III. However, there was an additive interaction between high-expression of DNMT1 mRNA and high-expression of MeCP2 mRNA in SCC or CIN II-III.</p><p><b>CONCLUSION</b>Results from our study revealed the fact that both high expression of DNMT1 protein and high expression of MeCP2 protein could increase the risk of cervix cancerization. According to our findings, there might be a synergistic action existed between DNMT1 and MeCP2 during the progression of cervix cancelation.</p>

Effects of gastrin on rat intestinal epithelial 1,25(OH)2D3-membrane associated rapid response steroid binding protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effects of gastrin on the expression of 1,25(OH)2D3-membrane associated rapid response steroid (1,25D3-MARRS) binding protein in rat intestinal epithelium.</p><p><b>METHODS</b>SD rats received intraperitoneal injections of gastrin, omeprazole or physiological saline. The protein expression of 1,25D3-MARRS binding protein in SD rat intestinal was determined with Western blotting and immunohistochemistry, and its mRNA levels determined by RT-PCR. The serum calcium and phosphate levels in the rats were also detected.</p><p><b>RESULTS</b>Immunohistochemistry showed that 1,25D3-MARRS binding protein was expressed mainly in the nuclei, cytoplasm and membrane of the intestinal epithelial cells. Both the protein and mRNA expression levels of 1,25D3-MARRS binding protein were up-regulated after treatments with gastrin and omeprazole (P<0.05), but the serum calcium and phosphate concentrations showed no obvious increase.</p><p><b>CONCLUSION</b>1,25D3-MARRS binding protein, which is widely expressed with versatile functionalities, is regulated by gastrin and shows high potentials in the study of gastrointestinal diseases.</p>

Effect of folate in modulating the expression of DNA methyltransferase 1 and methyl-CpG-bingding protein 2 in cervical cancer cell lines / 中华流行病学杂志

Objective To explore the effects of folate on the expression of DNA methyltransferase 1 (DNMT1) and methyl-CpG-bingding protein 2 (MeCP2) in cervical cancer cell lines.Methods Experimental study was carried out in vitro.Human cervical cancer cell lines,including C33A cell with HPV negative and Caski cell with HPV16 positive,were treated with different concentration of folate.The expression of DNMT1 and MeCP2 protein (by Western blot)and mRNA (by real-time PCR) were then detected in the two cell lines.Results It was found that supplement of folate was able to reduce the cell proliferation in C33A cell (r=0.984,P＜0.001) and Caski cell (r=0.978,P=0.002),as well as induced the cell apoptosis (C33A:r=0.989,P＜0.001 ;Caski:r=0.994,P＜0.001).Results showed that the expression levels of DNMT1 protein (C33A:r=-0.914,P＜ 0.001 ; Caski:r=-0.859,P=0.003) and MeCP2 protein (C33A:r=-0.830,P=0.005 ;Caski:r=-0.981,P＜0.001) decreased gradually with the increase of folate concentrations,but the expression of DNMT1 and MeCP2 mRNA was not observed in Caski or C33A cell.When at the same levels of folate,the expression of DNMT1 protein or mRNA was higher in Caski cell than in C33A cell.However,the expression of MeCP2 protein or mRNA was higher in C33A cell than in Caski cell.Conclusion Our fimding indicated that adequate foleta could effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,thus would reverse the aberration protein expression of DNMTl and MeCP2.That there might be a synergistic action between HPV16 infection and parafunction of DNMT l in cervical cancer,being noticed.

A survey of occupational health among polyether-exposed workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the occupational health of the workers simultaneously exposed to acrylonitrile, epoxyethane, epoxypropane, and styrene.</p><p><b>METHODS</b>A questionnaire survey was conducted in 70 front-line workers simultaneously exposed to acrylonitrile, epoxyethane, epoxypropane, and styrene (exposure group) and 50 managers (control group) in a polyether manufacturer; in addition, air monitoring at workplace and occupational health examination were also performed. The obtained data were analyzed.</p><p><b>RESULTS</b>The female workers in exposure group and the spouses of male workers in exposure group had significantly higher spontaneous abortion rates than their counterparts in control group (P < 0.01). The exposure group had a significantly higher abnormal rate of blood urea nitrogen than the control group (P < 0.01). The workers with different polyether-exposed working years had significantly higher mean levels of DNA damage than the control group (P < 0.01); the workers with not less than 5 and less than 20 polyether-exposed working years and those with not less than 20 polyether-exposed working years had significantly higher mean micronucleus rates than the control group (P < 0.01); there were no significant differences in overall chromosome aberration rate and mean level of DNA damage between each two groups of workers with different polyether-exposed working years (P > 0.05); the workers with not less than 5 and less than 20 polyether-exposed working years and workers with not less than 20 polyether-exposed working years had significantly higher mean micronucleus rates than those with less than 5 polyether-exposed working years (P < 0.01).</p><p><b>CONCLUSION</b>Simultaneous exposure to acrylonitrile, epoxyethane, epoxypropane, and styrene causes occupational hazards among the workers in polyether manufacturer.</p>