Pathologically triggered in situ aggregation of nanoparticles for inflammation-targeting amplification and therapeutic potentiation

Uncontrolled and persistent inflammation is closely related to numerous acute and chronic diseases. However, effective targeting delivery systems remain to be developed for precision therapy of inflammatory diseases. Herein we report a novel strategy for engineering inflammation-accumulation nanoparticles via phenolic functionalization. Different phenol-functionalized nanoparticles were first developed, which can undergo in situ aggregation upon triggering by the inflammatory/oxidative microenvironment. Phenolic compound-decorated poly (lactide-co-glycolide) nanoparticles, in particular tyramine (Tyr)-coated nanoparticles, showed significantly enhanced accumulation at inflammatory sites in mouse models of colitis, acute liver injury, and acute lung injury, mainly resulting from in situ cross-linking and tissue anchoring of nanoparticles triggered by local myeloperoxidase and reactive oxygen species. By combining a cyclodextrin-derived bioactive material with Tyr decoration, a multifunctional nanotherapy (TTN) was further developed, which displayed enhanced cellular uptake, anti-inflammatory activities, and inflammatory tissue accumulation, thereby affording amplified therapeutic effects in mice with colitis or acute liver injury. Moreover, TTN can serve as a bioactive and inflammation-targeting nanoplatform for site-specifically delivering a therapeutic peptide to the inflamed colon post oral administration, leading to considerably potentiated in vivo efficacies. Preliminary studies also revealed good safety of orally delivered TTN. Consequently, Tyr-based functionalization is promising for inflammation targeting amplification and therapeutic potentiation of nanotherapies.

Diagnostic Performance of Intestinal in Colorectal Cancer: A Meta-Analysis / 中华医学杂志(英文版)

<p><b>Background</b>Increasing evidence has supported the link of intestinal Fusobacterium nucleatum infection to colorectal cancer (CRC). However, the value of F. nucleatum as a biomarker in CRC detection has not been fully defined. In order to reduce the random error and bias of individual research, this meta-analysis aimed to evaluate the diagnostic performance of intestinal F. nucleatum in CRC patients and provide evidence-based data to clinical practice.</p><p><b>Methods</b>An article search was performed from PubMed, Embase, Cochrane Library, and Web of Science databases up to December 2017, using the following key words: "Fusobacterium nucleatum", "Fusobacterium spp.", "Fn", "colorectal cancer(s)", "colorectal carcinoma(s)", "colorectal neoplasm(s)", and "colorectal tumor(s)". Articles on relationships between F. nucleatum and CRC were selected according to the preestablished inclusion and exclusion criteria. This meta-analysis was performed using STATA 12.0 software, which included mapping of forest plots, heterogeneity tests, meta-regression, subgroup analysis, sensitivity analysis, and publication bias. The sensitivity, specificity, positive likelihood ratio (LR), negative LR, diagnostic odds ratio (DOR), and their corresponding 95% confidence interval (CI) of each eligible study were summarized.</p><p><b>Results</b>Finally, data for 1198 participants (629 CRC and 569 healthy controls) in 10 controlled studies from seven articles were included. The summary receiver operator characteristic curve was mapped. The diagnostic performance of intestinal F. nucleatum infection on CRC was as follows: the area under the curve: 0.86 (95% CI: 0.83-0.89), the pooled sensitivity: 0.81 (95% CI: 0.64-0.91), specificity: 0.77 (95% CI: 0.59-0.89), and DOR: 14.00 (95% CI: 9.00-22.00).</p><p><b>Conclusion</b>Intestinal F. nucleatum is a valuable marker for CRC diagnosis.</p>

Inflammation, insulin resistance and traditional chinese medicine treatment of diabetes mellitus / 中国药学杂志

Low grade inflammation has recently emerged as a common underlying cause of many chronic diseases. Heart disease, type 2 diabetes, Alzheimer's disease and many types of cancer have all been associated with chronic inflammation. Diabetes is a group of metabolic diseases characterized by hyperglycemia resulting from defects in insulin secretion, insulin action, or both. Insulin resistance is a key component in the development of diabetes mellitus which is caused by inflammation, and throughout the whole process of diabetes. In recent years, research on traditional Chinese medicine treatment of diabetes has got lots of achievements, and makes a further analysis about insulin resistance of diabetic, while has profound significance in the treatment of diabetes in the future. Insulin resistance can be improved with Chinese herb medicine using compound, single herbal drug and its extracts. This review focuses on the pathogenesis of inflammation and insulin resistance, and the effects of Chinese herb medicine on insulin resistance in diabetes.

A new bibenzyl compound from Dendrobium nobile / 药学学报

In this study, seven bibenzyl compounds were isolated from the stem of Dendrobium nobile by silica gel, MCI column chromatographic and preparative HPLC technology. By using spectroscopic techniques including NMR and MS, these compounds were identified as 4, α-dihydroxy-3, 5, 3'-trimethoxybibenzyl (1), 4, 5-dihydroxy-3, 3', α-trimethoxybibenzyl (2), 4, 4'-dihydroxy-3, 5, 3'-trimethoxybibenzyl (3), 4, 5-dihydroxy-3, 3'-dimethoxybibenzyl (4), 4, 3'-dihydroxy-3, 5-dimethoxybibenzyl (5), 5, 4'-dihydroxy-3, 3'-dimethoxybibenzyl (6) and 5, 3'-dihydroxy-3-methoxybibenzyl (7). Compound 1 is a new compound and compound 4 was isolated from this plant for the first time.

The influence of bFGF gene transfected BMSCs on inflammatory cytokines expression of COPD rat / 实用医学杂志

Objective To study the influence of bFGF gene transfected bone marrow-derived mesenchymal stem cells (BMSCs) on the inflammatory cytokines of COPD rat. Methods The BMSCs were separated from SD rat and cultured and then bFGF gene was imported to BMSCs by liposome transfection method. The samples were prepared into six groups: normal control group, COPD group (A), BMSCs group (B), pcDNA3.1-BMSCs group (C), bFGF-pcDNA3.1-BMSCs group (D), and bFGF group (E). The expressions of TNF-α and IL-1β by QRT-PCR were detected. Results Compared with COPD group, TNF-α and IL-1β genes from groups B to D dropped significantly (P 0.05). Conclusion BFGF transfected BMSCs, sample BMSCs and pcDNA3.1 transfected BMSCs can inhibit the expression of inflammatory cytokines of TNF-α and IL-1β, but there is no obvious advantage in comparison to bFGF transfected BMSCs and sample BMSCs in respect of inhibiting the expression of inflammatory cytokines of TNF-α and IL-1β.

Small bronchoscopic biopsy specimens for detecting egfr gene mutations in advanced lung adenocarcinoma / 第二军医大学学报

Objective To examine epidermal growth factor receptor (EGFR) gene mutations in small bronchoscopic biopsy specimens, so as to provide guidance for clinical targeted therapy. Methods Fifty-six female patients with advanced-stage III B-IV lung adenocarcinoma underwent endoscopic endobronchial biopsy of tumor tissues or transbronchial needle aspiration of mediastinal and hilar lymph nodes. Under the endoscope, 20 patients underwent only bronchial biopsy, 28 underwent only transbronchial needle aspiration (TBNA) lymph node biopsy, and 8 underwent both endobronchial biopsy and TBNA biopsy of mediastinal and hilar lymph nodes. A total of 64 specimens were collected and were subjected to detection of EGFR gene mutations after confirmation of lung adenocarcinoma. The specimens were then divided into endobronchial metastasis group and lymph node metastasis group, and the mutations of exons 19 and 21 were detected and the clinical efficacy of targeted therapy was analyzed. Results Exon 19 had higher positive rate in the endobronchial metastasis group (χ2=4.304,P=0.038), and exon 21 had higher positive rate in lymph node metastasis group (χ2=18.727,P=0.000). A total of 24 cases were included for the clinical efficacy assessments: 10 had endobronchial metastasis (exon 19 mutations in 8 cases, 21 exon in 2 cases), with the disease control rate being 90% (9/10) and median progression-free survival period being 14.8 months; 14 patients had lymph node metastasis (19 exon 3 cases, 21 exon 11 cases), with the disease control rate being 78.57% (11/14) and the median progression-free survival period being 9.2 months; the disease control rates were not significantly different between the two groups (P>0.05) and the median progression-free survival periods were significantly different between the two groups(χ2 = 4.134, P=0.042). Conclusion Mutations of different EGFR exons might relate to the metastasis forms of female advanced lung adenocarcinoma, with exon 19 prone to endobronchial metastasis and exon 21 to lymph node metastasis. Targeted therapy for patients with endobronchial metastasis has a better outcome than that for patients with lymph node metastasis.

Establishment of a cell model of insulin-resistant 3T3-L1 adipocytes / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes.</p><p><b>METHOS</b>Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10(-8), 10(-7), and 10(-6) mol·L(-1)) were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulin- resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1µmol·L(-1) DEX, and the changes of glucose concen- tration in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay.</p><p><b>RESULTS</b>Treatment of 3T3-L1 cells with DEX, IBMX and 10(-6) mol·L(-1)) insulin for 9 days resulted in the differentiation of >90% of the cells into mature adipocytes. IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 µmol·L(-1) DEX showed significantly increased glucose consumption (P=0.0003) as compared with the control group at 36 h (P<0.001).</p><p><b>CONCLUSION</b>3T3-L1 cells can be induced into mature adipocytes by exposure to 1 µmol·L(-1) DEX, 0.5 mmol·L(-1) IBMX and 10(-6) mol·L(-1)) insulin. A 96 h exposure to 1 µmol·L(-1) DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.</p>

Effects of rutaecarpine on inflammatory cytokines in insulin resistant primary skeletal muscle cells / 中国中药杂志

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.

Current prevalence rate of congenital heart disease in 12 month-old and younger infants among four regions of Guangdong province / 中华心血管病杂志

<p><b>OBJECTIVE</b>To summarize prevalence rate and region distribution of congenital heart disease (CHD) in 12-month-old and younger infants among four regions of Guangdong province, China.</p><p><b>METHODS</b>Data from Guangdong CHD monitoring network including 34 monitoring units covering different geographic regions were analyzed. Professional training on screening and diagnosing CHD was provided to each work group member to improve the diagnosis level. CHD infants under or aged 12 months detected in the monitoring unit were included. CHD was diagnosed by fetus and infants echocardiography.</p><p><b>RESULTS</b>From July 2004 to December 2010, 383 281 perinatal were registered and 3263 cases of CHD were detected in the 34 member units of Guangdong CHD monitoring network [total prevalence rate of CHD: 0.851% (3263/383 281), male prevalence rate: 0.868% (1799/207 347), female prevalence rate:0.828% (1456/175 843)].Stillbirth CHD prevalence rate was significantly higher than livebirth CHD prevalence rate [10.627% (676/6361) vs. 0.686% (2587/376 920), P < 0.01]. The total prevalence of CHD was significantly higher in Pearl River Delta region [0.906% (2826/311 823)] than in other regions [0.611% (437/71 458), P < 0.01]. Ventricular septal defect [39.93% (1033/2587) in livebirth] was the most dominant CHD, followed by patent ductus arteriosus [29.84% (772/2587)] and secundum atrial septal defect [13.76% (356/2587)].</p><p><b>CONCLUSIONS</b>The present data indicate that the prevalence of CHD in Guangdong is at the medium-upper level of the country associated with high stillbirth rate. The dominant type of CHD is ventricular septal defect. CHD prevalence is higher in the Pearl River Delta region than in other regions.</p>

Risk factors of congenital heart defects in fetal and infants born from 2004 to 2011 in Guangdong / 中华心血管病杂志

<p><b>OBJECTIVE</b>To analyze the risk factors of congenital heart defects (CHD) in fetal and infants born from 2004 to 2011 in Guangdong province.</p><p><b>METHODS</b>Babies with CHD aged from 28th week of gestation to 1 year old postnatal from July 1 2004 to December 31 2011 were registered in Guangdong CHD monitoring network with 34 participating units. Totally 2568 CHD cases were included, and 1: 1 matched with a normal control cohort by gender, living district and birth date (time span within 3 months).Exposed information of mother and father at pre-pregnancy and early pregnancy was collected. Post collinearity diagnostics analysis, univariate analysis results were included in a multivariate analysis model with forward stepwise conditional logistic regression.</p><p><b>RESULTS</b>Multivariate conditional logistic regression analysis showed that high risk factors for CHD included low birth weight infant (OR = 5.34, P < 0.01), macrosomia (OR = 1.67, P < 0.05), low per capita income (0-1200 yuan, OR = 1.68, P < 0.01), exposure to chemical agent at early pregnancy (OR = 19.72, P < 0.01), floating population (OR = 2.13, P < 0.01), abnormal reproductive history (OR = 3.18, P < 0.01), exposure to passive smoking (OR = 2.59, P < 0.01), suffering from fever (OR = 3.74, P < 0.01), equal to or more than twice parity (OR = 1.45, P < 0.01), living in a newly (within six months)-decorated-apartment (OR = 2.74, P < 0.01), suffering from virus infection (OR = 2.08, P < 0.01), rural residence (OR = 1.33, P < 0.01), living in an apartment within 50 meters of major traffic road (OR = 1.52, P < 0.01), syphilis infection at early pregnancy (OR = 13.06, P < 0.05) and father's drinking habit at pre-pregnancy (OR = 1.57, P < 0.05).</p><p><b>CONCLUSION</b>Numerous risk factors for CHD in fetal and infants of Guangdong province are indicated by our results, comprehensive intervention should be considered in pre-pregnancy and early pregnancy to reduce the risk of CHD.</p>

Effects of four kinds of Chinese medicine monomer on growth of PANC-1 xenograft tumor and studying of molecular mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>The antitumor effects of icarisid II, timosaponin A-III, neferine and salidroside were studied in PANC-1 xenograft tumor.</p><p><b>METHOD</b>To establish of the nude mice xenograft tumor model, PANC-1 cells were injected. When the tumor major diameter was reached 3-5 mm, the treatment was initiated. The mice were randomized into vehicle control and treatment groups of six animals per each. Chinese medicine monomer was injected intraperitoneally every day. In 23th day, mice were killed once a day, tumor tissue were isolated and weighed and divided into two parts. One part was fixed with formaldehyde for tissue section and immunohistochemistry, the another of tissue was frozen in liquid nitrogen then in - 80 degrees C refrigerator for gene and protein expression analysis.</p><p><b>RESULT</b>In PANC-1 tumor xenograft experiment, compared with model group, timosaponin A-III (1.0 mg x kg (-1)) exerted significant inhibitory effects on tumor growth. Timosaponin A-III suppressed mRNA expressions of VEGF (P < 0.05), reduced protein expressions of VEGF (P < 0.05), activated Caspase-3 protein. Icarisid II, neferine and salidroside had not an excelled antitumor effect.</p><p><b>CONCLUSION</b>Timosaponin A-III exerted an excelled antitumor effect. The antitumor mechanisms include anti-angiogenesis, apoptosis promotion.</p>

Study on the differences of risk factors regarding congenital heart defects between floating population and permanent residents in Guangdong / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the differences of risk factors on congenital heart defect (CHD)between floating population and permanent residents in Guangdong.</p><p><b>METHODS</b>A multicenter case-control study was carried out to investigate the risk factors of CHD in floating population and in permanent residents. Data was from 34 Guangdong CHD Monitoring Network centers during the year of 2004 to 2011. Exposed information related to the parents at pre-pregnancy and early pregnancy periods, was collected, using the same questionnaire survey methodology in the two populations. Possible risk factors were analyzed by univariate analysis and multivariate non-conditional logistic regression(ENTER method)methods. Risk factors were compared between the two populations.</p><p><b>RESULTS</b>Totally, 855 CHD cases and their controls from the floating population, as well as 1673 cases and their controls from the permanent residents were included in this study. Age of the children under study was defined from 28th week of gestation to 1 year old postnatal. In the floating population, specific risk factor for CHD appeared as:maternal passive smoking in early pregnancy, while the specific protective factor as high family income. However, the specific risk factors would include: having diseases as maternal diabetes mellitus or syphilis, living in a newly (within half a year) decorated house or with fetal macrosomia in the permanent residents. High education level showed as a risk factor in floating population, however contrarily, as protective factor to the permanent residents. Except for the factors related to having fever of the mother and infant with low birth weight, factors as having history of deliveries more than two, with maternal virus infection, exposure to chemical agent and negative bearing history etc., have higher OR values in floating population than in the permanent residents.</p><p><b>CONCLUSION</b>Significant differences of risk factors for CHD were noticed between floating population and the permanent residents, which have their individual specific risk factors. Most of the ORs appeared higher in floating population than in the permanent residents.</p>

Clinical analysis of bleeding during transbronchial needle aspiration / 第二军医大学学报

Objective To retrospectively analyze the incidence of bleeding during transbronchial needle aspiration (TBNA) procedures, so as to provide evidence for prevention and treatment of the condition. Methods TBNA procedures were performed in 178 patients with enlarged mediastinal and/or hilar lymph nodes, who were treated in the People's Hospital of Linyi. A total of 248 lymph node sites were punctured, including 100 for the groups of pretracheal and post-superior vena cava lymph nodes (4R), 10 for the groups of left paratracheal (aorta-pulmonary artery windows) lymph nodes (4L), 13 for the groups of pre-carinal lymph nodes (7),90 for the groups of sub-carinal lymph nodes (7), 12 for the groups of sub-subcarinal lymph nodes (7), 12 for the groups of right lung hilar lymph nodes (11R), and 11 for the groups of left lung hilar lymph nodes (11L).TBNA procedures were performed according to WANG's TBNA positioning and punctured method. The sites and incidence rates of bleeding sites during the procedures were analyzed. Results The highest incidence rate of bleeding (20%) was found during TBNA procedures in the sub-carinal lymph node groups (7), and the lowest incidence rate was found (2%) in the pretracheal and post-superior vena cava lymph node groups (4R).The incidence rates of bleeding were significantly different between different puncture sites as demonstrated by chi-square test (χ2=17.035,P=0.009). Conclusion Pretracheal and post-superior vena cava lymph nodes (4R) and sub-carinal lymph nodes (7) are the most common TBNA puncture position in the airway. TBNA procedure at sub-carinal lymph nodes (7) has the highest risk of bleeding. The massive bleeding during TBNA procedures can be avoided by prior enhanced CT and/or endobronchial ultrasound (EBUS)-TBNA of the lung.

Treatment of congenital hypertrophic pyloric stenosis with endoscopic pyloromyotomy / 中华儿科杂志

<p><b>OBJECTIVE</b>To evaluate the effect of the treatment of congenital hypertrophic pyloric stenosis (CHPS) with endoscopic pyloromyotomy.</p><p><b>METHOD</b>Nine consecutive infants (7 boys, 2 girls; age range 26 - 70 days; weight range 2.65 - 6.10 kg), with a diagnosis of CHPS according to typical clinical manifestations, transabdominal ultrasound (US), gastroenterography and gastroscope. All the cases had accompanying malnutrition, anaemia, metabolic alkalosis, and some were complicated with congenital heart disease. In gastroscope operating room, all the patients were given pentobarbital and midazolam intravenously. A gastroscope with an outer diameter of 5.9 mm was passed through mouth, stomach, pylorus to the descending segment of duodenum. Under gastroscopy, two incisions were made along the anterior and posterior wall of pylorus from the duodenal bulb to the antrum by using endoscopic electrosurgical needle knife and an arch sphincter sarcosome. Incisions were deepened by 2 to 3 procedures until the longitudinal muscle was exposed, about 2 to 4 mm according to transabdominal US performed before operation. The incision depth was 2 - 3 mm if pylorus wall was 4 - 6 mm in thickness; or 3 - 4 mm when the wall was thicker than 6 mm.</p><p><b>RESULT</b>The endoscope was easily passed through the pylorus to the duodenum post-operation. The transabdominal US and gastroenterography showed that liquid easily flew through pylorus. All patients were able to have regular feeding about 2 to 10 hours after the operation. Vomiting in all patients was significantly decreased in frequency and amount, and in 8 infants vomiting stopped within 1 week, in one case it did not stop until 1 month after the treatment. Some cases showed slight adverse reaction, no perforation or massive haemorrhage in stomach or intestines occurred in any of the patients during and post-operation. Eight infants were doing well at follow-up (range 2 to 9 months). One girl had recurred vomiting at normal feeding after a period of 1 month postoperation without vomiting. This case was cured by second endoscopic pyloromyotomy.</p><p><b>CONCLUSIONS</b>Endoscopic pyloromyotomy is effective, safe, simple, and offers several advantages: no need for open-abdomen surgery, feeding can be initiated rapidly.</p>

Association of hepatic lipase gene promoter polymorphism -514C/T with nonalcoholic fatty liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the prevalence of the hepatic lipase gene (LIPC) promoter polymorphism (at position -514) in patients with nonalcoholic fatty liver disease (NAFLD), and its relationship with the susceptibility to NAFLD.</p><p><b>METHODS</b>Genotype of LIPC promoter was detected with PCR-RFLP in 106 patients with NAFLD. Body mass index, waist-to-hip ratio (WHR), blood pressure, CHOL, HDL, LDL, TG, FPG and FINS of the patients were measured. Index of insulin resistance was determined using the homeostasis model assessment (HOMA) method. One hundred six healthy subjects matched for age and sex served as controls.</p><p><b>RESULTS</b>The frequency of CC genotype and C allele in the NAFLD group were significantly higher than those in the control group (31.1% vs 26.4%, 62.7% vs 54.2%, P<0.05). Compared with TT genotype, both CC genotype and CT genotypes had higher relative risk of NAFLD (OR: 3.73, 95% CI: 1.31, 10.63; OR: 3.60, 95% CI: 1.35, 9.60). At the same time, the non-carriers of T allele in -514 had higher WHR than the T carriers (0.877+/-0.06 vs 0.848+/-0.06, t=2.072, P<0.05)). Logistic regression analysis showed that T substitution in LIPC-514 position (OR: 1.28, 95% CI 0.10-0.74) had a lower susceptibility to NAFLD.</p><p><b>CONCLUSION</b>The LIPC-514C/T polymorphism is associated with WHR, and the T substitution of LIPC-514 may lower the susceptibility to NAFLD.</p>

The relationship between insulin resistance and adiponectin gene expression in nonalcoholic fatty liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship between insulin resistance (IR) and adiponectin gene expression in patients with nonalcoholic fatty liver disease (NAFLD).</p><p><b>METHODS</b>Subcutaneous (SC) and omental (OM) adipose tissues were obtained from 21 NAFLD patients with obesity (n=10) and nonobesity (n=11) and also from 24 subjects (without NAFLD) with obesity (n=11) and nonobesity (n=13) who served as controls. Adiponectin mRNA expression levels in subcutaneous and omental adipose tissues were measured using SYBR Green I quantitative real-time PCR. The levels of plasma adiponectin and insulin were measured with ELISA. IR was estimated using the homeostasis assessment (HOMA).</p><p><b>RESULTS</b>The scores of HOMA-IR levels of the NAFLD patients and the controls with obesity and nonobesity were: 3.0+/-0.8, 2.8+/-0.9, 2.0+/-0.6, 1.2+/-0.5 respectively. The relative mRNA expression of adiponectin and blood adiponectin levels in NAFLD patients differed significantly from those of the controls. The HOMA-IR negatively correlated with the adiponectin mRNA expression levels of adipose tissues (r = -0.5) and blood adiponectin; it positively correlated with body mass index (r = 0.4), waist-hip-ratio (r = 0.4) and serum triglyceride (r = 0.3), but did not correlate with serum total cholesterol (r = 0.2).</p><p><b>CONCLUSION</b>IR of NAFLD patients was linked to low adiponectin gene expression in their adipose tissues. This finding suggests that low adiponectin gene expression may play a role in the pathogenesis of insulin resistance and NAFLD.</p>

Relationship between leptin gene of adipose tissues and nonalcoholic fatty liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate leptin mRNA expressions in subcutaneous (SC) and omental (OM) adipose tissues of patients with nonalcoholic fatty liver disease (NAFLD), and their relationships with insulin resistance (IR), blood leptin, blood triglyceride, total blood cholesterol, blood glucose, body weight index and waist-hip ratio.</p><p><b>METHODS</b>SC and OM adipose tissues were obtained from 10 obese and 11 nonobese NAFLD patients and from 11 obese and 13 nonobese patients without NAFLD, who served as controls. Leptin mRNA expression levels in the subcutaneous and omental adipose tissues were measured using SYBR Green I quantitative real-time PCR. IR was estimated using homeostasis assessment (HOMA). The levels of plasma leptin and insulin were measured using ELISA.</p><p><b>RESULTS</b>The relative mRNA expression of leptin, HOMA-IR and blood leptin levels in NAFLD differed significantly from those of the controls (P < 0.05). The leptin/GAPDH ratio of the obese and nonobese NAFLD and control cases were 1.32 +/- 0.12, 0.99 +/- 0.05, 1.10 +/- 0.09, 0.87 +/- 0.13 respectively. The expression levels of SC and OM adipose leptin mRNA in NAFLD patients were positively correlated with HOMA-IR (r=0.72, P < 0.05), blood leptin (r=0.69, P < 0.05), blood triglyceride (r=0.32, P < 0.05), body weight index (r=0.57, P < 0.05) and waist-hip ratio (r=0.50, P <0.05).</p><p><b>CONCLUSION</b>The primary reason for high levels of blood leptin is high leptin mRNA expression in adipose tissues; in both obese and nonobese patients with NAFLD; high levels of blood leptin and the leptin mRNA expression in adipose tissues and IR exist. These findings suggest that leptin resistance exists in patients with NAFLD and leptin resistance is positively correlated with NAFLD, the same as in insulin resistance.</p>

nm23-H1 gene inhibits lung cancer cell invasion through down-regulation of PKC signal pathway / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the molecular mechanisms of nm23-H1 for regulating PKC signal pathway before and after transfection with nm23-H1 gene.</p><p><b>METHODS</b>Using Western-blot, Boyden-chamber, MTT and laser scanning confocal microscopy (LSCM) techniques to detect the distribution of PKC in cytosol and plasma membrane, changes of invasion and proliferation activity, PKC translocation status and changes of intracellular Ca(2+) concentration among different human pulmonary carcinoma cells with transfected or untransfected nm23-H1 gene, and changes of the three cell lines after treatment with Calphostin C, a PKC inhibitor.</p><p><b>RESULTS</b>(1) The expression of PKCalpha, PKCbeta II on L9981 and L9981-pLXSN cell membrane, which was in activated status, was remarkably higher than those in L9981-nm23-H1 cell line (P < 0.001). The expression of PKCalpha, PKCbeta II in cytosol in L9981 and L9981-pLXSN cell lines, which was in inactivated status, was lower than those in L9981-nm23-H1 cell line (P < 0.001). It means that the PKC signal pathway was activated in L9981 and L9981-pLXSN cell lines. (2) PKCalpha and PKCbeta II mainly located in nuclei and perinuclear area in L9981 and L9981-pLXSN cells, which were in active status, and the Ca(2+) concentration in these cells was obviously higher than that in L9981-nm23-H1 cell line (P < 0.01). In L9981-nm23-H1 cell line, which was transfected with nm23-H1 gene, PKCalpha and PKCbeta II mainly located in soluble cytosolic section, in an inactive status. (3) The invasion and proliferation ability of L9981 and L9981-pLXSN lung cancer cells was higher than that of L9981-nm23-H1 cell line (P < 0.001). There was no statistically significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05). (4) After treated with PKC inhibitor Calphstin C, the expression of PKC and PKCbeta II in membrane in L9981 and L9981-pLXSN cell lines was down-regulated (P < 0.001), PKCalpha and PKCbeta II were mainly located in cytosolic area, mainly in an inactive status, and the Ca(2+) concentration was found to be decreased in all the three cell lines. The invasion and proliferation ability of the three lung cancer cell lines were obviously decreasing (P < 0.001). However, the invasion and proliferation ability of L9981-nm23-H1 lung cancer cell line was still lower than that of L9981 and L9981-pLXSN lung cancer cell lines (P < 0.001). There was also no significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05).</p><p><b>CONCLUSION</b>The results of this study suggest that nm23-H1 gene might inhibit the invasion and metastasis of lung cancer cells by down-regulating PKC signaling pathway. The Ca(2+) in cells might be involved in this process.</p>

Transfection of tumor metastasis suppressor gene nm23-H1 can up-regulate the activity of GSK-3β in human high-metastasis large cell lung cancer cell line L9981 / 中国肺癌杂志

<p><b>BACKGROUND</b>To investigate the influence of tumor metastasis suppressor gene nm23-H1 on the activity of glycogen synthase kinase 3β (GSK-3β) in human high-metastasis large cell lung cancer cell line L9981.</p><p><b>METHODS</b>The levels of GSK-3β expression in cytoplasm and nucleus were determined with anti- GSK-3β antibody in human high-metastasis large cell lung cancer cell line L9981 (cell line with nm23-H1 gene deletion), L9981-nm23-H1 (cell line with nm23-H1 transfected) and L9981-pLXSN (cell line with vector transfected) by Western blot method. The activity of GSK-3β among those three cell lines was detected by immunoprecipitation and analysed by a radioactive isotope scintillation counter before and after treating with 20 mmol/L LiCl.</p><p><b>RESULTS</b>(1) The expression indensity of GSK-3β of cytoplasm and nucleus was (6 341±541) and (4 356±490) IOD in L9981-nm23-H1, (3 613±383) and (705±75) IOD in L9981-pLXSN, and (3 736±298) and (675±57) IOD in L9981, respectively. A high significance in GSK-3β expressive indensity of both cytoplasm and nucleus existed among L9981-nm23-H1, L9981-pLXSN and L9981 (P < 0.01); Multiple comparison: A highly significant difference was observed when L9981-nm23-H1 was compared with L9981-pLXSN or L9981 (P < 0.01), but no significant difference was observed between L9981-pLXSN and L9981 (P > 0.05). (2) The GSK-3β activity of cytoplasm and nucleus was (28 955±2 509) and (9 247±924) CPM in L9981-nm23-H1, (11 241±1 495) and (1 492±176) CPM in L9981-pLXSN, and (12 505±1 469) and (1 763±125) CPM in L9981, respectively. A highly significant difference in GSK-3β activity of both cytoplasm and nucleus existed among L9981-nm23-H1, L9981-pLXSN and L9981 (P < 0.01); Multiple comparison: the GSK-3β activity in L9981-nm23-H1 was significantly higher than that in L9981-pLXSN and L9981 (P < 0.01), but no significant difference was observed between the L9981-pLXSN and L9981 (P > 0.05). (3) After treatment with 20 mmol/L LiCl, the expressive indensity of GSK-3β of cytoplasm and nucleus was (4 718±549) and (3 823±350) IOD in L9981-nm23-H1, (2 030±155) and (217±15) IOD in L9981-pLXSN, and (2 164±151) and (224±19) IOD in L9981, respectively. No significant difference in GSK-3β expressive indensity existed between before and after treatment with LiCl in L9981-nm23-H1 (P > 0.05). However, the GSK-3β expressive indensity in cytoplasm and nucleus before treatment was remarkably higher than those after treatment in both L9981-pLXSN and L9981 (P < 0.05). (4) After treatment with 20 mmol/L LiCl, the GSK-3β activity in cytoplasm and nucleus was (11 099±1 112) and (3 748±215) CPM in L9981-nm23-H1, (4 447±430) and (1067±159) CPM in L9981, and (4 435±427) and (909±156) CPM in L9981-pLXSN, respectively. The GSK-3β activity both in cytoplasm and nucleus after treatment with LiCl was remarkably lower than that before treatment in L9981-nm23-H1, L9981-pLXSN and L9981 (P < 0.01 or P < 0.05).</p><p><b>CONCLUSIONS</b>(1) Transfection of nm23-H1 gene can significantly up-regulate the expression level and activity of GSK-3β in human high-metastasis large cell lung cancer cell line L9981; (2) LiCl can remarkably suppress the upregulation effects of nm23-H1 gene on GSK-3β activity in L9981 cell line; (3) The effects of nm23-H1 gene on suppressing the signal transduction of Wnt pathway might be carried out through upregulating GSK-3β expression and activity in human high-metastasis large cell lung cancer cell line L9981.</p>

Study on protein kinase C translocation before and after transfection of nm23-H1 gene in human lung cancer cells using Laser scanning confocal microscope / 中国肺癌杂志

<p><b>BACKGROUND</b>To explore the influences of nm23-H1 gene transfection and protein kinase C (PKC) inhibitor Calphostin C on PKC signal transduction pathway in human high-metastasis large cell lung cancer cell line L9981, and to evaluate the effects of nm23-H1 gene on translocation and activation in subcellular region.</p><p><b>METHODS</b>The translocation of PKC in subcellular region was observed in L9981 before and after nm23-H1 gene transfection and Calphostin C treatment by Laser scanning confocal microscope (LSCM) method.</p><p><b>RESULTS</b>PKC-α and PKC-βII were found to locate in different subcellular site in L9981 before and after nm23-H1 gene transfection. PKC-α and PKC-βII mainly located in nucleus and perinucleus in L9981 and L9981-pLXSN cell lines, which were in active status. PKC-α and PKC-βII mainly located in soluble cytosolic fraction in L9981-nm23-H1 cell line and were inactive status. PKC-α and PKC-βII mainly located in cytosolic fraction and were in inactive status in all the three cell lines after treatment with Calphostin C.</p><p><b>CONCLUSIONS</b>The results suggest that nm23-H1 gene might make PKC to translocate from nucleus and perinucleus to soluble cytosolic fraction in L9981 cell line. PKC inhibitor, Calphostin C, can also make PKC to translocate from nucleus and perinucleus to soluble cytosolic fraction in L9981, L9981-pLXSN cell lines. Both transfection of nm23-H1 gene and treatment with Calphostin C can suppress the PKC signal transduction in L9981 cell line.</p>