Acupuncture at "umbilical four-acupoints" for chronic insomnia and its comorbid symptoms / 中国针灸

OBJECTIVE@#To observe the effects of acupuncture at "umbilical four-acupoints" on chronic insomnia and its comorbid symptoms.@*METHODS@#A total of 120 patients with chronic insomnia were randomly divided into an observation group (60 cases, 8 cases dropped off) and a control group (60 cases, 5 cases dropped off). The patients in the observation group were treated with acupuncture at regular acupoints (Baihui [GV 20] and bilateral Shenmen [HT 7], Neiguan [PC 6], Anmian [Extra]) and "umbilical four-acupoints", while the patients in the control group were treated with acupuncture at regular acupoints. Acupuncture was given once a day, 6 times a week, for a total of 3 weeks in the two groups. The Pittsburgh sleep quality index (PSQI), insomnia severity index (ISI) scores were observed before treatment, after treatment and in follow-up of one month after treatment completion; the Beck anxiety inventory (BAI), Beck depression inventory (BDI), fatigue severity scale (FSS), and Epworth sleepiness scale (ESS) scores were observed before and after treatment; the sleep parameters of polysomnography (PSG), including sleep latency (SL), awake-up time (AT), sleep efficiency (SE) and total sleep time (TST), were observed before and after treatment using polysomnography monitor in the two groups.@*RESULTS@#Compared with those before treatment, the PSQI and ISI scores in both groups were reduced after treatment and in follow-up (P<0.05), and the PSQI and ISI scores in the observation group were lower than those in the control group after treatment and in follow-up (P<0.05). Compared with those before treatment, the BAI, BDI, FSS and ESS scores in both groups were reduced after treatment (P<0.05), and the BAI, BDI, FSS and ESS scores in the observation group were lower than those in the control group after treatment (P<0.05). Compared with those before treatment, the SL and AT in both groups were reduced after treatment (P<0.05), while SE and TST were increased after treatment (P<0.05); after treatment, the SL and AT in the observation group were lower than those in the control group (P<0.05), while SE and TST in the observation group were higher than those in the control group (P<0.05).@*CONCLUSION@#On the basis of regular acupoint selection, acupuncture at "umbilical four-acupoints" could improve sleep quality, alleviate the severity of insomnia, and improve the comorbid symptoms i.e. anxiety, depression, fatigue and lethargy in patients with chronic insomnia.

Association between serum vitamin D level and immune imbalance in advanced schistosomiasis patients with liver fibrosis / 中国血吸虫病防治杂志

Objective To examine the relationship between serum vitamin D level and immune imbalance in advanced schistosomiasis patients with liver fibrosis. Methods A total of 120 advanced schistosomiasis patients with liver fibrosis that were admitted to the Department of Schistosomiasis of The First Hospital of Jiaxing City from May 2016 to September 2018 were recruited as the observation group, and 50 healthy volunteers randomly sampled from the hospital during the same period served as the control group. The serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ cell proportion, CD8+ cell proportion, 25-hydroxyvitamin D [25(OH)D] levels were compared between the two groups. Liver fibrosis was classified into grade I, II and III according to the classification criteria of liver fibrosis by ultrasonography, and the serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ proportion, CD8+ proportion, 25(OH)D levels were compared among patients with grade I, II and III liver fibrosis. In addition, all patients were classified into the sufficient group, the insufficient group and the deficient group according to the serum vitamin D level, and the serum IgG antibody, IgA antibody, C3 complement, C4 complement, CD4+ proportion, CD8+ proportion, 25(OH)D levels were compared among these three groups. Moreover, the associations of the serum vitamin D level with these immune indicators were examined. Results The 120 advanced schistosomiasis patients with liver fibrosis included 58 men and 62 women, and had a mean age of (72.00 ± 3.00) years. There were 32 cases with grade I liver fibrosis, 46 cases with grade II liver fibrosis, and 42 cases with grade III liver fibrosis. There were no significant differences between the observation group and the control group in terms of serum D-dimer, total cholesterol (TC), triglyceride (TG), C3 complement or C4 complement levels (t = 2.467, 0.322, 0.790, -2.432 and -2.630, all P values > 0.05); however, there were significant differences seen in alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood calcium, blood phosphorus, IgG antibody, IgA antibody, CD4+ proportion, CD8+ proportion, and 25(OH)D levels (t = 5.130, 6.382, -1.341, 2.361, 8.708, 11.783, -2.995, -6.543 and -3.022, all P values < 0.05). In addition, there were significant differences in AST, ALT, blood phosphorus, IgA antibody, C3 complement, CD8+ cell proportion and 25(OH)D levels among patients with grades I, II and III liver fibrosis (F = 19.704, 16.254, 62.669, 49.347, 5.430, 5.434 and 5.783, all P values < 0.05). There were significant differences in ALT, blood phosphorus, IgA antibody, CD8+ cell proportion and 25(OH)D levels between patients with grades I and III liver fibrosis (all P values < 0.05), and significant differences were seen between patients with grades II and III liver fibrosis in terms of blood phosphorus, IgA antibody and CD8+ cell proportion (all P values < 0.05), while there was a significant difference in the CD8+ cell proportion between patients with grades I and II liver fibrosis (P < 0.05). Moreover, there were significant differences among the sufficient, insufficient and deficient groups in terms of IgG antibody, IgA antibody, C3 complement, CD4+ cell proportion and CD8+ cell proportion (F = 13.303, 59.623, 8.698, 9.969 and 12.805, all P values < 0.05), and there was a significant difference in the CD8+ cell proportion between the insufficient and deficient groups (P < 0.05). Pearson correlation analysis revealed that serum 25(OH)D level were negatively associated with IgG and IgA antibody levels (r = -0.754 and -0.773, both P values < 0.05), and positively associated with C3 complement, CD4+ cell proportion and CD8+ cell proportion in advanced schistosomiasis patients with liver fibrosis (r = 0.827, 0.850 and 0.830, all P values < 0.05). Conclusion Immune imbalance occurs in advanced schistosomiasis patients with liver fibrosis, and serum vitamin D level may correlate with immune imbalance in advanced schistosomiasis patients with liver fibrosis.

Effect of Duhuo Jisheng Tang on PERK/Bip Signaling Pathway in Knee Osteoarthritis Model Rats / 中国实验方剂学杂志

Objective: To explore the possible mechanism of Duhuo Jisheng Tang in relieving knee osteoarthritis based on protein kinase R-like endoplasmic reticulum kinase (PERK)/immunoglobulin-binding protein (Bip) signaling pathway.Method: A model of knee osteoarthritis was established by cold stimulation.Rats were randomly divided into blank group,model group,celecoxib group (0.021 g·kg-1),low,medium and high-dose Duhuo Jisheng Tang groups (8.37,16.72,33.48 g·kg-1).Blank group and model group were given equal volume of physiological saline.The changes of knee joint diameter were recorded.The pathological changes of rat articular cartilage were observed by hematoxylin-eosin (HE) staining.The expressions of tumor necrosis factor-alpha (TNF-α),interleukin-1β(IL-1β) and hyaluronic acid (HA) in serum were detected by enzyme-linked immunosorbent assay (ELISA).The mRNA and protein expression levels of PERK,Bip and cysteinyl as parates pecific protein-9(Caspase-9) in cartilage were detected by Real-time PCR and Western blot.Result: The knee joint redn ess and the joint diameter of celecoxib group and high-dose Duhuo Jisheng Tang group were improved,and the joint diameter was reduced significantly (Pα,IL-1β and HA were increased in model group (PPPα,IL-1β and HA in serum of celecoxib group and high-dose Duhuo Jisheng Tang group were decreased (PPPPConclusion: Duhuo Jisheng Tang can alleviate the symptoms of knee osteoarthritis model rats,and its mechanism may be related to the regulation of PERK/Bip signaling pathway in rat cartilage.

The regulation of retinoid X receptor-mediated oxidative stress pathway in rat pulmonary ischemia/reperfusion injury / 生理学报

The aim of this study was to investigate the regulatory role of retinoid X receptor (RXR)-mediated oxidative stress pathway in rat pulmonary ischemia/reperfusion injury (PIRI) and the underlying mechanism. Seventy-seven male Sprague-Dawley (SD) rats were randomly divided into 7 groups (n = 11): control group, sham group, sham+9-cis-retinoid acid (9-cRA, RXR agonist) group, sham+HX531 (RXR inhibitor) group, ischemia/reperfusion (I/R) group, I/R+9-cRA group, and I/R+HX531 group. The unilateral lung I/R model was established by obstruction of left lung hilus for 30 min and reperfusion for 180 min in vivo. The rats in I/R+9-cRA and I/R+HX531 groups were given intraperitoneal injection of 9-cRA and HX531 before thoracotomy. After reperfusion, the left lung tissue was taken to evaluate the lung tissue injury, and the oxidative stress-related indexes of the lung tissue were detected by the corresponding kits. The lung tissue morphology and the ultrastructure of the alveolar epithelial cells were observed by HE staining and transmission electron microscope, respectively. The protein expression of RXR in lung tissue was observed by immunofluorescence labeling method, and the expression level of nuclear factor E2-related factor (Nrf2) protein was detected by Western blot. The results showed that, compared with the sham group, the I/R group exhibited obviously injured lung tissue, decreased SOD activity, increased MDA content and MPO activity, and down-regulated expression level of Nrf2 protein. Compared with the I/R group, the I/R+9-cRA group showed alleviated lung tissue injury, increased activity of SOD, decreased MDA content and MPO activity, and up-regulated expression levels of RXR and Nrf2 protein. The above-mentioned improvement effects of 9-cRA were reversed by HX531 treatment. These results suggest that RXR activation can effectively protect the lung tissue against I/R injury, and the mechanism may involve the activation of Nrf2 signaling pathway, the enhancement of antioxidant level and the reduction of oxidative stress response.

Serum vitamin D expression in advanced schistosomiasis patients with hepatic fibrosis and its association with disease progression / 中国血吸虫病防治杂志

Objective To compare the expression of serum vitamin D in advanced schistosomiasis patients with grade I and II hepatic fibrosis, and to preliminarily examine its associations with the internal diameter of the main portal vein and progression of hepatic fibrosis. Methods The medical records of 126 advanced schistosomiasis patients with grade I and II hepatic fibrosis referred to Jiaxing First Hospital from March 2012 to September 2015 were retrospectively analyzed. The internal diameter of the main portal vein and serum 25-hydroxyvitamin D3 [25(OH)D3] level were measured. The progression of hepatic fibrosis was followed up, and the serum vitamin D level was compared between patient with disease progression and stable disease. Results The 126 advanced schistosomiasis patients included 72 men and 54 women, and had ages of 62 to 80 years. There were 58 cases with grade I hepatic fibrosis and 68 cases with grade II hepatic fibrosis. There were significant differences between patients with grade I and II hepatic fibrosis in terms of hemoglobin, white blood cell count, prothrombin time, international normalized ratio, activated partial thromboplastin time, fibrinogen or 25(OH)D3 level (all P > 0.05), and significant differences were seen in alanine aminotransferase, aspartate aminotransferase, blood calcium, blood phosphorus levels and the internal diameter of the main portal vein (all P values < 0.05). In addition, a lower serum 25(OH)D3 level was detected in patients with broadened internal diameter of the main portal vein than in those with normal internal diameter of the main portal vein [(19.08 ± 1.36) nmol/L vs. (25.61 ± 6.69) nmol/L, P < 0.05]. Following 3-year follow-up, there were 73 cases with progression of hepatic fibrosis, and a significantly lower serum vitamin D level was found in patients with disease progression than in those with stable disease [(20.00 ± 0.81) nmol/L vs. (25.47 ± 5.91) nmol/L, P < 0.05]. Conclusions Vitamin D deficiency is common in advanced schistosomiasis patients with hepatic fibrosis, and it may be associated with the internal diameter of the main portal vein and the progression of hepatic fibrosis disease.

Associations between circulating irisin, opg and gestational diabete mellitus / 医学研究生学报

Objective In recent decades, the morbidity of gestational diabetes mellitus (GDM) has been increased dramatically. Researches have found irisin can improve islet function, however there are few researches on opg and gestational diabetes. This study aimed to investigate the changes of circulating irisin, opg levels in GDM patients and their correlation with insulin resistance (IR), pancreatic β-cell function to explore the potential mechanism of GDM.Methods We enrolled newly diagnosed GDM women (GDM group, n=70) and normal glucose tolerance women (NGT group, n=70) who undertook regular prenatal checkups in Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from May to December 2015. HbA1c, HOMA-IR, insulin sensitivity, HOMA-β, early insulin secretion index(EISI, ΔI60/ΔG60), serum irisin and opg levels of all participants were tested. Correlation analysis were performed between serum irisin and opg levels and each index to analyze GDM risk factors.Results The fasting serum opg level in GDM group ([2224.76±881.47]pg/mL) was significantly higher than that in NGT group([1148.15±700.13]pg/mL) and the fasting serum irisin level in GDM group ([417.26.76±113.75]ng/mL) was significantly lower than that in NGT group([530.62±150.76]ng/mL) , both the differences were of statistical significance(P<0.05). HOMA-IR, OGTT AUCg and AUCi in GDM group were significantly higher than those in NGT group, while insulin sensitivity and ΔI60/ΔG60 index were significantly lower than those in NGT group (P<0.05). The fasting serum irisin level was in negative correlation with HOMA-IR and OGTT AUCg(r=-0.179, -0.258, P<0.05) and in positive correlation with insulin sensitivity(r=0.212, P<0.05); the opg level was in positive correlation with HOMA-IR(r=0.219, P<0.05) and in negative correlation with insulin sensitivity and ΔI60/ΔG60 index (r=-0.290, 0.183, P<0.05). Logistic regression analysis showed irisin (OR=0.994, 95%CI: 0.990~0.998) and opg (OR=5.802, 95%CI: 2.835~11.873) were the influencing factors of GDM (P<0.05).Conclusion Irisin and opg have certain correlation with IR and β-cell function. Either the decrease of serum irisin level or the increase of opg level may increase the risk of GDM.

Effects of low-intensity pulsed ultrasound on fracture healing and caveolin-1 gene expression in the radius defects of rabbits / 复旦学报（医学版）

Objective To explore the effect of low-intensity pulsed ultrasound (LIPUS) on long bone fracture healing and to examine caveolin-1 gene expression in the radius defects of rabbits.Methods A total of 24 New Zealand rabbits with 3-mm bone defects at lower 1/3 in both radii were randomly assigned to 4 groups (n=6).Daily LIPUS treatment was performed to the right fracture sites at a intensity of 30 mW/cm2 for 20 minutes,while the left sites received sham treatment with power off.To assess the effects of LIPUS on bone defects,X-ray imaging and hematoxylin-eosin staining were applied 7,14,21,28 days after the surgery.Additionally,the immunohistochemical staining was used to determine the subcllular localization of caveolin-1 and semi-quantify the caveolin-1 level,qPCR was performed to detect the mRNA level of caveolin-1,gene Col2a1 and Col10a1,and osteocalcin.Results On day 14,the radiological score of the right radii and mineralized callus area were significantly higher than that of the left ones,both of them were elevated with time flied.Histological examination suggested that the differentiation and apoptosis of chondrocytes along with the formation and bridging of the bone trabeculas appeared earlier in the right radius defects.The immunohistochemical staining showed that on day 7 and 14,the level of caveolin-1 increased with the proliferation and differentiation of condrocytes,and was significantly higher in callus tissues on the right sites.On day 21 and 28,the mesenchymal stem cells migrated to the surface of cartilage matrix started to differentiate into osteoblasts,the level of caveolin-1 decreased,and was significantly lower on the right sites.The result of qPCR indicated that compared with the left sites the caveolin-1 gene expression on the right sites was significantly higher on day 7,while significantly lower on day 21.The mRNA expression levels of Col2a1,Col10a1 and osteocalcin on the right sites were significantly higher on day 7 and 14,but they were significantly lower on day 21 and 28,except for Col10a1 on day 28.Conclusions Advancing endochondral ossification is considered to be a crucial mechanism during long bone fracture healing promoted by LIPUS.The caveolin-1 gene expression first increased in the chondrocytes then decreased in the mesenchymal stem cells during the process.

Role of TRPC6 in pulmonary artery smooth muscle cells proliferation and apoptosis under hypoxia and hypercapnia / 生理学报

The present study was to investigate the role of TRPC6 in pulmonary artery smooth muscle cells (PASMCs) proliferation and apoptosis under hypoxia and hypercapnia. PASMCs were isolated from chloral hydrate-anesthetized male Sprague-Dawley (SD) rats. Cellular purity was assessed by immunofluorescence staining for smooth muscle α-actin under fluorescence microscopy. Passage 4-6 PASMCs were starved for 24 h in serum-free DMEM and divided into 5 groups randomly: normoxia, hypoxia and hypercapnia, DMSO, TRPC6 inhibitor SKF-96365 and TRPC6 activator OAG groups. The normoxic group was incubated under normoxia (5% CO, 21% O, 37 °C) for 24 h, and the others were incubated with corresponding drugs under hypoxic and hypercapnic (6% CO, 5% O, 37 °C) atmosphere for 24 h. TRPC6 mRNA was detected by reverse transcription-PCR. TRPC6 protein was detected by Western blotting. The proliferation of PASMCs was performed by CCK-8 kit. Apoptosis of the PASMCs was detected using TUNEL assay. The [Ca]in the PASMCs was measured using Fura 2-AM fluorescence. The results showed that the expressions of TRPC6 mRNA and protein, and [Ca]were upregulated under hypoxic and hypercapnic conditions. Hypoxia and hypercapnia promoted cellular proliferation and inhibited apoptosis in the PASMCs. OAG enhanced the above-mentioned effects of hypoxia and hypercapnia, whereas SKF-96365 reversed these effects. These results suggest that TRPC6 may play a role in PASMCs proliferation and apoptosis under hypoxia and hypercapnia by regulating [Ca].

Inductive effect of zinc oxide nanoparticles on interleukin 8 gene expression in human bronchial epithelial cells and its regulatory mechanism / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To clarify the effect of zinc oxide nanoparticles (ZnO-NPs) (30 nm in diameter) on the interleukin 8 (IL-8) gene expression in human bronchial epithelial cells (BEAS-2B) and its regulatory mechanism.</p><p><b>METHODS</b>BEAS-2B cells were used in the study. The MTT assay was employed to evaluate the damage to BEAS-2B cells by ZnO-NPs. RT-PCR and ELISA were used to measure the mRNA and protein expression levels of IL-8 in the BEAS-2B cells exposed to ZnO-NPs. The IL-8 mRNA decay assay was used to determine the effect of ZnO-NPs on IL-8 mRNA stability.</p><p><b>RESULTS</b>Exposure to ZnO-NPs significantly increased the level of IL-8 mRNA in BEAS-2B cells and the level of IL-8 protein in supernatant medium. The transcription inhibitor significantly reduced the mRNA expression of IL-8 induced by ZnO-NPs. ZnO-NPs significantly delayed IL-8 mRNA degradation in the BEAS-2B cells that were pretreated with actinomycin D for terminating IL-8 mRNA synthesis.</p><p><b>CONCLUSION</b>ZnO-NPs can increase the mRNA and protein expression levels of IL-8 and IL-8 mRNA stability in BEAS-2B cells.</p>

Coal tar pitch smoke extract-induced pyroptosis in human bronchial epithelial cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore whether coal tar pitch smoke extract (CTP) induced pyroptosis in human bronchial epithelial cells (BEAS-2B).</p><p><b>METHODS</b>BEAS-2B cells were treated with different concentrations of CTP (1, 3 µg/ml) for 8h and 24 h, respectively. Lactic dehydrogenase (LDH) activity and interleukin-1 beta (IL-1β) levels in the supernatants of cell culture media were measured with LDH activity or human IL-1β ELISA kit, respectively. The activity of Caspase-1 was measured with Caspase-1 colorimetric assay kit.</p><p><b>RESULTS</b>The activity of caspase-1 in 1 and 3 µg/ml CTP groups were (9.29 ± 0.30) and (8.67 ± 0.59) µmol/ml respectively which were both significantly increased compared to that (7.42 ± 0.59) µmol/ml in the control group (P < 0.05) after 8 h exposure, but there was no significant difference in the activity of LDH and levels of IL-1β in the cell culture media among the CTP and control groups. 24 h after exposure, the activity of LDH in the CTP (1, 3 µg/ml) groups were (1323.03 ± 28.53) and (1148.45 ± 16.42) U/dl respectively which were significantly higher than that (1091.93 ± 26.64) U/dl in the control group (P < 0.05), and the levels of IL-1β in the CTP (1 and 3 µg/ml) groups were (125.37 ± 25.00) pg/ml and (92.04 ± 19.09) pg/ml respectively which were significantly higher than that (46.20 ± 14.43) pg/ml in the control group (P < 0.05), but there was no significant difference in the activity of Caspase-1 among CTP and control groups (P < 0.05).</p><p><b>CONCLUSION</b>CTP treatment induced early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels, indicative of pyroptosis in human bronchial epithelial cells.</p>

CT-Guided Core Needle Biopsy of Deep Suprahyoid Head and Neck Lesions

OBJECTIVE: To evaluate the efficacy of computer tomography (CT)-guided core needle biopsy (CNB) in the diagnosis of deep suprahyoid lesions in patients with treated head and neck cancers. MATERIALS AND METHODS: Between December, 2003 and May, 2011, 28 CT-guided CNBs were performed in 28 patients with deep suprahyoid head and neck lesions. All patients had undergone treatment for head and neck cancers. Subzygomatic, paramaxillary, and retromandibular approaches were used. The surgical results, response to treatment, and clinical follow-up were used as the diagnostic reference standards. RESULTS: All biopsies yielded adequate specimens for definitive histological diagnoses. A specimen from a right parapharyngeal lesion showed atypia, which was deemed a false negative diagnosis. Diagnostic accuracy was 27/28 (96.4%). Two minor complications were encountered: a local hematoma and transient facial palsy. Between the 18 or 20 gauge biopsy needles, there was no statistical difference in the diagnostic results. CONCLUSION: CT-guided core needle biopsy, with infrequent and minor complications, is an accurate and efficient method for the histological diagnosis of deep suprahyoid lesions in post-treated head and neck cancer patients. This procedure can preclude an unnecessary surgical intervention, especially in patients with head and neck cancers.

Changes of centrosome and related protein in malignant transformation of BEAS-2B cell induced by coal tar pitch smoke extracts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze the centrosome abnormalities in the malignant transformation of human bronchial epithelial cells (BEAS-2B) induced by coal tar pitch smoke extracts and to investigate the role and action mechanism of centrosome in the lung cancer induced by coal tar pitch.</p><p><b>METHODS</b>Medium-temperature coal tar pitch smoke extracts were used to treat immortalized human bronchial epithelial cells (BEAS-2B) and establish a malignant transformation model. The treated BEAS-2B cells were used as exposure group, and solvent control group and normal control group were also set for passage culture. The changes of centrosome in BEAS-2B cells seeded on coverslips were evaluated by indirect immunofluorescence assay. The mRNA expression of p53, p21, and cyclin E in BEAS-2B cells was measured by real-time quantitative RT-PCR, and their protein levels in BEAS-2B cells seeded on coverslips were measured by semiquantitative immunohistochemical analysis.</p><p><b>RESULTS</b>The overall rate of centrosome abnormalities in BEAS-2B cells at passage 20 was 6.56±1.01% in the exposure group, significantly higher than those in the normal control group (3.40±0.86%) and solvent control group (3.14±0.59%) (P < 0.05). In addition, the exposure group had a significantly higher overall rate of centrosome abnormalities in BEAS-2B cells at passage 30 compared with the normal control group and solvent control group (22.39±9.5% vs 4.34±1.04%, P < 0.05; 22.39±9.5% vs 4.33±1.20%, P < 0.05). Compared with the normal control group and solvent control group, the exposure group had significantly decreased mRNA and protein expression of p53 and significantly increased mRNA and protein expression of cyclin E in BEAS-2B cells at passages 20 and 30 (P < 0.05).</p><p><b>CONCLUSION</b>Centrosome abnormalities occur before the malignant transformation in BEAS-2B cells treated with coal tar pitch smoke extracts, and they may be mediated by the p53/p21/cyclin E signaling pathway.</p>

Effects of heat shock protein 70-2 gene polymorphisms on the transcription of HSP 70-2 mRNA and the translation of HSP 70 protein in lung cancer / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study aimed to investigate the effects of gene polymorphism of heat shock protein 70-2 (HSP 70-2) 1267A/G on the mRNA level HSP 70-2 mRNA and the protein level HSP 70 in human lung cancer.</p><p><b>METHODS</b>Forty six lung cancer patients diagnosed histopathologically between February and August 2008 from a hospital in zhengzhou were enrolled as the subjects in this study. Gene polymorphism of HSP 70-2 1276A/G in 46 patients with lung cancer was detected by PCR-RFLP. The mRNA levels of HSP 70-2 mRNA and the protein levels of HSP 70 in lung tissue and para-cancerous tissues of these subjects were determined by RT-PCR and Western blotting respectively.</p><p><b>RESULTS</b>The expression levels of HSP 70-2 mRNA (1.02 ± 0.30) and HSP 70 protein (0.44 ± 0.12) in the lung cancer tissues was significantly higher than that in para-cancerous tissues (0.19 ± 0.04, 0.12 ± 0.02). The relative levels of HSP 70-2 mRNA in the subjects with AA genotype (1.32 ± 0.22) were significantly higher than the patients with AG genotype or GG genotype (0.95 ± 0.17, 0.70 ± 0.16) at the site of 1267 (A/G) (P < 0.01); however, the relative protein levels of HSP 70 were 0.47 ± 0.13 (AA genotype), 0.42 ± 0.11 (AG genotype), 0.45 ± 0.11 (GG genotype), respectively, which showed no statistically significant difference (P > 0.05).</p><p><b>CONCLUSION</b>The polymorphism of HSP 70-2 1267 (A/G) is highly associated with the transcription level of HSP 70-2 mRNA, but not with the expression level of HSP 70 protein.</p>

The polymorphisms of XRCC1 gene and susceptibility to pulmonary cancer / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the relationship between the polymorphisms of DNA repair gene XRCC1 and susceptibility to pulmonary cancer.</p><p><b>METHODS</b>A case-control study of 209 lung cancer patients and 256 control subjects was conducted to investigate the role of XRCC1 gene in lung cancer. Genotyping was performed using PCR based restriction fragment length polymorphism (PCR-RFLP) technique.</p><p><b>RESULTS</b>The frequency (19.1%) of XRCC1-194 Trp/Trp in case group was significantly higher than that (10.9%) in control group (P < 0.05), OR for lung cancer was 2.215 (95% CI: 1.276-3.845). The frequency (6.7%) of XRCC1-280 His/His in case group was significantly higher than that (4.3%) in control group (P < 0.05), OR for lung cancer was 2.46 (95% CI: 1.141-5.304). There was no significant difference for XRCC1-399 Gln/Gln genotype between the two groups. Interaction analysis of gene polymorphisms and environment factors indicated that there was interactions between XRCC1-194 Trp/Trp and XRCC1-280 His/His genotypes and smoking. The risks of lung cancer in smokers with XRCC1-194 Arg/Trp+Trp/Trp and XRCC1-280 His/His+Arg/His were 4.889 (95% CI: 2.828-8.452) and 6.281(95% CI: 3.572-11.046), respectively.</p><p><b>CONCLUSION</b>These findings supported the hypothesis that the interaction of polymorphisms of XRCC1-194 Trp/Trp, XRCC1-280 His/His with smoking resulted in the increased risk of lung cancer, and the polymorphisms of XRCC and smoking could play an role in development of lung cancer.</p>

The effects of monocyte-macrophages on malignant transformation of human bronchial epithelial cells induced by extracts from coal tar pitch / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of monocyte-macrophages (THP-1) in malignant transformation of human bronchial epithelial cells (BEAS-2B) cells induced by coal tar pitch (CTP) and the expression of TNF-α in the process of the cell malignant transformation.</p><p><b>METHODS</b>BEAS-2B cells and THP-1 Cells were divided into four groups: coal tar pitch (CTP) group, benzo(a)pyrene [B(a)P] group, dimethyl sulfoxide (DMSO) group, BEAS-2B and THP-1 co-culture (co-culture group) group. Carcinogenesis model was established. The soft agar colony formation, chromosome aberrations and cell cycle tests were used to detect the cellular malignant transformation. The ELISA assay was utilized to measure the levels of TNF-α in the supernatant of CTP group and co-culture group.</p><p><b>RESULTS</b>The chromosome number abnormalities could be observed in early stage of the experiment (the 10th generation cells), which showed the increased ratio of aneuploid to polyploid, and the decreased number of diploid. The colony formation rate of co-culture group (the 20th generation cells) was 17.63‰ ± 0.97‰, which was significantly higher than that (13.94‰ ± 0.84‰) of CTP group and that (12.96‰ ± 1.62‰) of B(a)P group (P < 0.05). The proportion of S phase cells in the co-culture group was 44.49% ± 0.68%, which was significantly higher than that (38.19% ± 1.26%) of CTP group and that (36.41% ± 1.19%) of B(a)P group (P < 0.05). The TNF-α level in the co-culture group was significantly higher than that in CTP group (P = 0.001).</p><p><b>CONCLUSION</b>Monocyte-Macrophages can accelerate the malignant transformation of BEAS-2B cells induced by CTP and increase the expression level of TNF-α.</p>

DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats.</p><p><b>METHODS</b>The primary macrophages and fibrocytes of SD rats were co-cultured directly and indirectly, which were exposed to SiO2 at the doses of 25, 50 and 100 g/ml. The transdifferentiation of cFb was identified with immunohistochemical assay. The genomic DNA methylation levels of cFb were detected with HPLC.</p><p><b>RESULTS</b>Under the condition of indirect co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 19.9%, 26.9% and 30.3%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 22.0% (P < 0.05). Under the condition of ThinCert(TM) direct co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 22.2%, 30.2% and 36.7%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 20.6% (P < 0.05).</p><p><b>CONCLUSION</b>Under the co-culture condition in vitro, SiO2 could reduce the genomic DNA methylation levels of cFb. The ThinCert(TM) direct co-culture can be used to study the silicosis fibrosis.</p>

The change of telomere protein in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>By testing the changes of telomere binding protein in malignant transformation BEAS-2B cells induced by coal tar pitch smoke extracts, to study the role of protection of telomeres 1 (POT1), telomeric repeat binding factor 1 (TRF1) and TRF2 in tumorgenesis that contact with coal tar pitch.</p><p><b>METHODS</b>The BEAS-2B cells were induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, the protein expression variations were determined by cell culture overslip of immunohistochemical methods.</p><p><b>RESULTS</b>In malignant transformation cells, the mRNA expression level (POT1: 0.63 ± 0.04, TRF1: 0.36 ± 0.01) and the protein expression level (POT1: 0.36 ± 0.05, TRF1: 0.09 ± 0.03) of POT1 and TRF1 was statistically significant decreased compared to that of BEAS-2B group (mRNA: POT1: 1.00 ± 0.04, TRF1: 1.01 ± 0.16; protein: POT1: 0.55 ± 0.07, TRF1: 0.27 ± 0.07) and DMSO group (mRNA: POT1: 0.89 ± 0.12, TRF1: 0.90 ± 0.08; protein: POT1: 0.55 ± 0.10, TRF1: 0.26 ± 0.04) (P < 0.05); mRNA expression level (1.45 ± 0.07) and the protein expression level (0.88 ± 0.06) of TRF2 was increased compared to that of BEAS-2B group (mRNA: 1.00 ± 0.07, protein: 0.48 ± 0.06) and DMSO group (mRNA: 1.00 ± 0.06, protein: 0.50 ± 0.06) (P < 0.05).</p><p><b>CONCLUSION</b>The change of gene and protein expression level in POT1, TRF1, and TRF2 involved in the process that evolved into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.</p>

Involvement of Toll-like receptor in silica-induced tumor necrosis factor alpha release from human macrophage cell line / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To characterize the role of Toll-like receptor 4 (TLR4) in silica-induced production of tumor necrosis factor alpha (TNFalpha) from macrophage cell line.</p><p><b>METHODS</b>The human macrophage cell line THP-1 was incubated with silica suspension. Cell media were collected and TNFalpha levels in the supernatants measured with ELISA. To examine the involvement of TLR4 in silica-induced TNFalpha release, the neutralizing antibody (HTA125) against human TLR4 receptor was employed to pretreat THP-1 cells prior to silica treatment. Moreover, murine macrophages expressing wild type or mutated TLR4 were also treated with silica to verify the effect of TLR4 in silica-induced TNFalpha release.</p><p><b>RESULTS</b>Compared with the control group [(3.18 +/- 0.41) pg/ml], the TNFalpha release in cells exposed to 100 microg/ml silica for 4 h and 8 h [(4.71 +/- 0.84), (6.22 +/- 0.58) pg/ml, respectively] increased 1.48 and 1.96 fold, respectively. Pretreatment of THP-1 cells with 20 microg/ml HTA125 antibody significantly blocked silica-induced TNFalpha release by 27%. Furthermore, the TNFalpha content released from cells expressing mutated TLR4 reduced by 30% in compared with that from the cells expressing wild type TLR4 after silica stimulation.</p><p><b>CONCLUSION</b>TLR4 mediates silica-induced TNFalpha release from macrophages.</p>

Change of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin expression in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>to study the role of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin in tumorgenesis that contact with coal tar pitch.</p><p><b>METHODS</b>the BEAS-2B cells was induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, and the protein expression variation were determined by cell culture overslip of immunohistochemical methods.</p><p><b>RESULTS</b>in malignant transformation cells, the mRNA and the protein expression level of SMC1 gene was not statistically significantly different compared with the BEAS-2B group and DMSO group (P > 0.05); SMC3 and Separase was increased and Securin was decreased (P < 0.05), while the difference between other two control groups was not significant (P > 0.05).</p><p><b>CONCLUSIONS</b>the up expression level of SMC3 and Separase and the down expression level of Securin are involved in the process that evolves into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.</p>

Construction of lentiviral mediated CyPA siRNA and its functions in non-small cell lung cancer / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.</p><p><b>METHODS</b>First, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.</p><p><b>RESULTS</b>It was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Lentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.</p>