The clinical significance of the expression of XB130 in gastric cancer tissues / 新乡医学院学报

Objective To investigate the relationship between the expression of XB130 and the histodifferentiation,lymph node metastasis and survival time of patients with gastric cancer.Methods Seventy-two specimens of gastric cancer tissues and seventy-two specimens of paracancerous tissues were collected from June 2011 to June 2012 in Henan Province People's Hospital.The expression of XB130 protein in gastric cancer and paracancerous tissues was detected by immunohistochemistry.The expression of XB130 mRNA in gastric cancer and paracancerous tissues was detected by real-time quantitative polymerase chain reaction.The relationship between the expression of XB130 and clinicopathological features of gastric cancer was analyzed.Results There were 26 cases of positive expression of XB130 protein and 46 cases of negative expression of XB130 protein among the 72 cases of gastric cancer tissues,and the positive expression rate was 36.1％ (26/72).There were 52 cases of positive expression of XB130 protein and 20 cases of negative expression of XB130 protein among the 72 cases of paracancerous tissues,and the positive expression rate was 72.2％ (52/72).The positive expression rate of XB130 protein in gastric cancer tissues was significantly lower than that in paracancerous tissues (x2 =16.200,P ＜ 0.05).The positive expression rate of XB130 protein in poorly and moderately differentiated gastric cancer tissues was significantly lower than that in well differentiated gastric cancer tissues (x2 =5.786,P ＜0.05).The positive expression rate of XB130 protein in gastric cancer tissues with lymph node metastasis was significantly lower than that in gastric cancer tissues without lymph node metastasis (x2 =4.281,P ＜0.05).The relative expression of XB130 mRNA in gastric cancer tissues and paracancerous tissues was 1.52 ±0.46 and 2.28-± 0.51 respectively,the expression of XB130 mRNA in gastric cancer tissues was significantly lower than that in paracancerous tissues (t =-21.744,P ＜0.05).The expression of XB130 mRNA in well differentiated gastric cancer tissues was significantly higher than that in poorly and moderately differentiated gastric cancer tissues (t =-13.982,P ＜ 0.05).The expression of XB130 mRNA in gastric cancer tissues with lymph node metastasis was significantly lower than that in gastric cancer tissues without lymph node metastasis (t =-19.906,P ＜ 0.05).The mean survival time in patients with high and low expression of XB130 mRNA was (37.040 ± 14.826) and (21.529 ± 11.789) months respectively,the survival time in patients with high expression of XB130 mRNA was significantly higher than that in patients with low expression of XB130 mRNA (t =9.121,P ＜0.05).Conclusion XB130 may be involved in the occurrence and development of gastric cancer,and it is associated with the differentiation,lymph node metastasis and survival time of patients with gastric cancer.

Huaiqihuang Granules () reduce proteinuria by enhancing nephrin expression and regulating necrosis factor κB signaling pathway in adriamycin-induced nephropathy / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Huaiqihuang Granules (, HQH), a mixture of Chinese herbs including Trametes robiniophila Murr, Fructus Lycii and Polygonatum sibiricum, on adriamycininduced nephropathy (ADRN) in rats and its underlying mechanisms.</p><p><b>METHODS</b>Rats with ADRN were divided into four groups: the sham group, the model group (distilled water), the low-dose HQH-treated (2 g/kg) group, and the high-dose HQH-treated (4 g/kg) group. Body weight and 24-h urinary protein (Upro) were checked every week. After 5-week intervention, at the end of the study, the rats were sacrificed and blood samples were collected for examination of biochemical parameters, including glomerular morphological makers, podocyte shape, cellular apoptosis, expressions of nephrin, inflammatory and apoptosis markers.</p><p><b>RESULTS</b>HQH ameliorated the rat's general status, proteinuria, renal morphological appearance and glomerulosclerosis. The decreased expression of nephrin in ADRN rats was increased by HQH, as well as the impaired podocyte foot process fusion. Cytosolic levels of p65 and inhibitor of nuclear factor κBα (IκBα) were decreased in ADRN rats, and recovered by the treatment of HQH. Consistently, the induced expression of tumor necrosis factor α (TNF-α), phosphorylated nuclear factor κB p65 (p-NFκB p65) and IκBα in ADRN were markedly suppressed by HQH. In addition, induction of Bax, cleaved caspase-3 and cytochrome C in ADRN rats were suppressed by HQH, indicating the amelioration of apoptosis.</p><p><b>CONCLUSION</b>HQH could ameliorate renal impairments in ADRN rats by increasing nephrin expression, inhibiting NF-κB signaling pathway via the down-regulation of p-NF-κB p65 and p-IκBα, and suppression of glomerular and tubular apoptosis.</p>

Biomechanical analysis of the correlation between sacral tilt displacement and L₅S₁ disc degeneration / 中国骨伤

<p><b>OBJECTIVE</b>To biomechanical analysis of the correlation between sacral tilt displacement and L₅-S₁ disc degeneration.</p><p><b>METHODS</b>From July 2011 to July 2013, 81 patients with lumbar disc herniation and sacroiliac joint disorder including 45 males and 36 females with an average age of (45.39±1.30) years ranging from 18 to 65 years old were selected. The course of the disease ranged from 1 to 144 months with an average of (12.64±2.19) months. All patients were taken lumbar spine lateral X-ray films, the lumbar curvature angle, L₄-L₅ or L₅-S₁ intervertebral gap distance between points, and the lumbosacral angle was measured and correlated analyzed.</p><p><b>RESULTS</b>The lumbar curvature of female patients with L₅S₁DH were significantly larger than male patients [(22.18±8.62)° vs (16.17±4.97)°,<0.05]. Lumbar curvature and lumbosacral angle showed a positive correlation in LDH (=0.48,<0.01,y=7.25+0.38x,<0.01); Male patients with L₄-₅DH were more obvious (=0.55,<0.05, y=5.80+0.43x,<0.01); Female patients with L₅S₁DH were particularly evident(=0.74,<0.01,y=0.91x-5.30,<0.01). The lumbosacral angle and L₄-₅ intervertebral gap was a positive correlation in L₄-₅DH(=0.27,<0.05); While L₅-S₁ intervertebral gap and lumbosacral angle were not correlated(>0.05) in L₅S₁DH.</p><p><b>CONCLUSIONS</b>The sacral tilt displacement and L₅-S₁ disc degeneration were closely related to provide a new understanding philosophy and therapeutic approach for clinical treatment of intractable lumbar L₅S₁DH.</p>

Study on fluorescence PCR-melting curve method for detecting CYP2C9 and VKORC1 gene polymorphism in Xinjiang Hui population / 国际检验医学杂志

Objective To use the fluorescence PCR‐melting curve method to detect CYP2C9 and VKORC1 gene polymorphism in Xinjiang Hui population ,to analyze their gene distribution and gene mutation frequency ,and to evaluate the clinical applicability of the fluorescence PCR‐melting curve method .Methods The fluorescence PCR‐melting curve method and sequencing method were adopted to contrastively detect CYP2C9*2 ,CYP2C9*3 and VKORC1(‐1639G/A)gene polymorphism .Results Among detected 228 Xinjiang Hui individuals ,199 cases of CYP2C9*1/*1 ,2 cases of CYP2C9*1/*2 ,26 cases of CYP2C9*1/*3 and only 1 case of CYP2C9*3/*3 were detected ,no case of CYP2C9*2/*2 and CYP2C9*2/*3 was detected .Two kinds of allele G and A were detected for VKORC1(‐1639G/A) ,in which VKORC1‐1639G/G type was detected in 2 cases ,VKORC1‐1639G/A type was detected in 39 cases and VKORC1‐1639A/A type was detected in187 cases ,compared with the sequencing method ,the results of the fluorescence PCR‐melting curve method were completely consistent .Conclusion Xinjiang Hui population also has CYP2C9 gene *2 ,*3 loci and VKORC1 gene(‐1639G/A) locus polymorphism ,their occurrence frequency has a certain difference with Xingjiang Uygur and other regional populations ,the adopted fluorescence PCR‐melting curve method used in the gene polymorphism detection can meet clinical detection requirements .

Molecular mechanism of emodin on inhibiting autophagy induced by HBSS in renal tubular cells / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the regulative effects and possible mechanisms of emodin on autophagy induced by starvation in rat's renal tubular epithelial cells (NRK-52E).</p><p><b>METHOD</b>Firstly, Hank's balanced salt solution (HBSS) was used to induce starvation and the protein expression of microtubule-associated protein 1 light chain 3 (LC3) I/II, an autophagic marker of mammalian congener, was detected by Western blot with or without the treatment of emodin. Secondly, the changes of red fluorescent protein-microtubule associated protein light chain3 (RFP-LC3) fluorescent particles, treated by HBSS (1 mL) and bafilomycin A1 (10 nmol x L(-1)) with or without emodin, were observed through fluorescence microscopy in NRK-52E cells transient transfected by RFP-LC3 plasmid. With the intervention of mammalian target of rapamycin mTOR inhibitor rapamycin (100 nmol x L(-1)) , the effect of blocking mTOR signaling pathway on autophagic inhibition of emodin was observed. Finally, the effect of mTOR signaling pathway on autophagic inhibition of emodin was further evaluated through the over-expression of endogenous mTOR inhibitory protein DEP domain-containing mTOR-interacting protein-(DEPTOR).</p><p><b>RESULT</b>HBSS hunger could induce high protein expression of LC3 II in NRK-52E cells, and the intervention of emodin could reverse the unregulated protein expression of LC3 II induced by HBSS. The number of RFP-LC3 fluorescent particles was increased after the co-treatment of HBSS and bafilomycin A1, and this increase was inhibited by emodin. After the co-treatment of rapamycin, emodin and HBSS, the LC3 II protein expression restored in NRK-52E cells, compared with the treatment of HBSS. Over-expression of DEPTOR could also block the inhibitive effect of emodin on LC3 II protein expression.</p><p><b>CONCLUSION</b>Emodin could inhibit HBSS-induced LC3 II protein expression and the activation of autophagy in NRK-52E cells, and the effect of blocking autophagy may be mediated through mTOR signaling pathway.</p>

Clinical implication of urinary protein markers in diabetic nephropathy and interventional effects of Chinese herbal medicine / 中国中药杂志

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.

Molecular mechanism of rhein on inhibiting autophagic protein expression in renal tubular epithelial cells via regulating mTOR signaling pathway activation / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effects and molecular mechanisms of rhein on reducing starvation-induced autophagic protein expression in renal tubular epithelial ( NRK-52E) cells.</p><p><b>METHOD</b>Hank's balanced salt solution (HBSS) was used to induce NRK-52E cells to be in the state of starvation. After the intervention of HBSS for 0, 0.5,1, 2 and 6 hours, firstly, the protein expression of microtubule-associated protein 1 light chain 3(LC3 I/II), which is a key protein in autophagy, was detected. Secondly, the protein expressions of mammalian target of rapamycin (mTOR) and phosphorylated-mTOR Ser2448 (p-mTOR S2448) were examined. And then, after the co-treatment of rhein (5 mg x L(-1)) and HBSS (1 mL) without or with mTOR inhibitor, rapamycin (100 nmol x L(-1)), the protein expressions of LC3 I/II, mTOR and p-mTOR S2448 were tested, respectively.</p><p><b>RESULT</b>HBSS could induce the up-regulation of LC3 II and the down-regulation of p-mTOR S2448 at protein expression level in NRK-52E cells. The co-treatment of rhein and HBSS could reversely regulate the protein expressions of LC3 II and p-mTOR S2448 in NRK-52E cells significantly. The co-treatment of rapamycin, rhein and HBSS could recover the level of LC3 II protein expression in HBSS-intervened NRK-52E cells.</p><p><b>CONCLUSION</b>HBSS induces autophagy in renal tubular epithelial cells by inhibiting mTOR signaling pathway activation. Rhein reduces the autophagic protein expression in renal tubular epithelial cells through regulating mTOR signaling pathway activation, which is the possible effects and molecular mechanisms.</p>

Mechanisms of cordycepin on improving renal interstitial fibrosis via regulating eIF2α/TGF-β/Smad signaling pathway / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects and mechanisms of cordycepin,an effective component of cordyceps militaris, on renal interstitial fibrosis (RIF) and its related eIF2α/TGF-β/Smad signaling pathway.</p><p><b>METHOD</b>Firstly, 15 C57BL/6 mice were randomly divided into 3 groups,the control group (Group A), the model group (Group B) and the cordycepin-treated group (Group C). After renal interstitial fibrotic model was successfully established by unilateral ureteral obstruction (UUO), the mice in Group C were intraperitoneally administrated with cordycepin(5 mg x kg(-1) d(-1)) and the ones in Group A and B were administrated with physiological saline for 5 days. At the end of the study, the obstructed kidneys were collected and detected for the pathological changes of RIF, and the mRNA expressions of collagen type I (Col I) and α-smooth muscle actin (α-SMA) in the kidney by Northern blot. Secondly, after renal tubular epithelial (NRK-52E) cells cultured in vitro were exposed to transforming growth factor (TGF) -β with or without cordycepin, the mRNA expressions of Col I and collagen type IV( Col IV) by Northern blot, and the protein expressions of eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α ( p-eIF2α), Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) were tested by Western blot.</p><p><b>RESULT</b>In vivo, cordycepin alleviated RIF in model mice, including improving fibrotic pathological characteristics and mRNA expressions of Col I and α-SMA. In vitro, cordycepin induced the high expression of p-elF2α, and inhibited the expressions of p-Smad2/3, Col I and Col IV induced by TGF-β in NRK-52E cells.</p><p><b>CONCLUSION</b>Cordycepin attenuates RIF in vivo and in vitro, probably by inducing the phosphorylation of eIF2α, suppressing the expression of p-Smad2/3, a key signaling molecule in TGF-β/Smad signaling pathway, and reducing the expressions of collagens and α-SMA in the kidney.</p>

Characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy and its regression analysis with traditional Chinese medicine symptom / 中国中药杂志

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.

Chronic Kidney Disease and Associated Cardiovascular Risk Factors in Chinese with Type 2 Diabetes

BACKGROUND: To determine the frequency of chronic kidney disease (CKD) and its associated risk factors in Chinese type 2 diabetic patients, we conducted a cross-sectional study in Nanjing, China, in the period between January 2008 and December 2009. METHODS: Patients with type 2 diabetes under the care by Jiangsu Province Official Hospital, Nanjing, China were invited for assessment. CKD was defined as the presence of albuminuria or estimated glomerular filtration rate or =30 mg/g. RESULTS: We recruited 1,521 urban Chinese patients with type 2 diabetes (mean age, 63.9+/-12.0 years). The frequency of CKD and albuminuria was 31.0% and 28.9%, respectively. After adjusted by age and sex, hypertension, anemia and duration of diabetes were significantly associated with CKD with odds ratio (95% confidence interval) being 1.93 (1.28 to 2.93), 1.70 (1.09 to 2.64), and 1.03 (1.00 to 1.06), respectively. CONCLUSION: In conclusion, CKD was common in the urban Nanjing Chinese with type 2 diabetes. Strategies to prevent or delay progression of kidney disease in diabetes should be carried out at the early disease course of type 2 diabetes.

Validity of Glycated Hemoglobin in Screening and Diagnosing Type 2 Diabetes Mellitus in Chinese Subjects

BACKGROUND/AIMS: The application of glycated hemoglobin (HbA1c) for the diagnosis of diabetes is currently under extensive discussion. In this study, we explored the validity of using HbA1c as a screening and diagnostic test in Chinese subjects recruited in Nanjing, China. METHODS: In total, 497 subjects (361 men and 136 women) with fasting plasma glucose (PG) > or = 5.6 mmol/L were recruited to undergo the oral glucose tolerance test (OGTT) and HbA1c test. Plasma lipid, uric acid, and blood pressure were also measured. RESULTS: Using a receiver operating characteristic curve, the optimal cutoff point of HbA1c related to diabetes diagnosed by the OGTT was 6.3%, with a sensitivity and specificity of 79.6% and 82.2%, respectively, and the area under the curve was 0.87 (95% confidence interval, 0.83 to 0.92). A HbA1c level of 6.5% had a sensitivity and specificity of 62.7% and 93.5%, respectively. When comparing the HbA1c > or = 6.5% or OGTT methods for diagnosing diabetes, the former group had significantly higher HbA1c levels and lower levels of fasting and 2-hour PG than the latter group. No significant difference was observed in the other metabolism indexes between the two groups. CONCLUSIONS: Our results suggest that HbA1c > or = 6.5% has reasonably good specificity for diagnosing diabetes in Chinese subjects, which is in concordance with the American Diabetes Association recommendations.

Effect of cinobufagin on nuclear factor-kappaB pathway in HepG2 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of cinobufagin on nuclear factor-kappaB (NF-kappaB) pathway of liver cancer cell line HepG2.</p><p><b>METHODS</b>Dual-luciferase cis-reporting system was used to detect the relative value of pNF-kappaB-TA-luc upon tumor necrosis factor-alpha (TNF-alpha) stimulation of NF-kappaB pathway. Western blotting was used to detect the protein level of NF-kappaB p65, and RT-PCR was used to detect the gene transcription level of intercellular adhesion molecule-1 (ICAM-1), a target downstream gene of NF-kappaB.</p><p><b>RESULTS</b>At the concentration of 0.25 and 0.5 microg/ml, cinobufagin significantly lowered the relative value of luciferase (P<0.05). The results of Western blotting showed that cinobufagin significantly suppressed the protein expression of NF-kappaB p65. The transcription level of ICAM-1 was reduced by different doses of cinobufagin.</p><p><b>CONCLUSION</b>The anti-cancer effect of cinobufagin may be related to its activity in inhibiting the activation of NF-kappaB pathway.</p>

Antibacterial activity of recombinant thanatin expressed by E.coli ER2566 / 南方医科大学学报

<p><b>OBJECTIVE</b>To express antibacterial peptide thanatin in the prokaryotic expression system and test its antibacterial activity.</p><p><b>METHODS</b>The DNA sequence coding for the 21 peptides of thanatin was synthesized using the preferred genetic codes of E. coli, cloned into pTYB11 plasmid, and transformed into E.coli ER2566. The expression of thanatin fused with intein was induced by IPTG in E.coli, and intein-thanatin specifically bound to the column through intein tag was cleaved overnight at 4 degrees celsius; in DTT/cysteine buffer.</p><p><b>RESULTS</b>The cleaved thanatin was eluted with a protein concentration of 245 microg/ml in the first 4 ml. The purified thanatin had showed strong antibacterial activities against G- bacteria such as Shigella flexneri, Klebsiella pneumoniae, Shigella snnei, Escherichia coli O157, toxin producing Escherichia coli, Pseudomonas aeruginosa, and fungi such as Candida albicans, with especial potency in killing drug-resistant Klebsiella pneumoniae, Pseudomonas aeruginosa, and extended-spectrum beta-lactamases (ESBL)-producing E.coli. Eighty strains of drug-resistant (ESBL-producing) and 30 strains of sensitive E. coli were used for anti-bacterial assay, and no significant differences in the antibacterial activity of thanatin were found between the sensitive and drug-resistant E. coli (P>0.05).</p><p><b>CONCLUSION</b>The recombinant thanatin obtained shows strong antibacterial activity against drug-resistant and sensitive bacteria, and can be a potential substitute for routine antibiotics in the treatment of G- bacterial infections.</p>

Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito / 病毒学报

An artificial intron consisting of the 5'-donor site (from the first intron of the human beta-globin gene) and the 3'-acceptor site (from the intron of an immunoglobulin gene heavy chain variable region) was obtained with a splice overlap extension PCR and was then inserted in frame into the coding sequence of nostructural protein NS1 gene fused to GFP gene in a recombinant mosquito densovirus plasmid p7NS1-GFP. The constructed plasmid was named as p7NS1-Intron-GFP. The plasmid p7NS1-Intron-GFP was co transfected with the helper plasmid pUCA into C6/36 cells, then the packaged recombinant and wild type viruses were purified and recovered. The second-instars of Aedes albopictus larvae were exposed to recombinant and wild type virus mixed stock. The high level GFP expression in C6/36 cells and larvae was observed under fluorescence microscope, indicating that the inserted artificial intron exerted its normal function in self-splicing both in vitro and in vivo. This study laid a foundation for application of an artificial intron in insect cells and development of new strategy for genetic engineering technology of mosqtuito and its pathogens.

Isolation, identification and analysis of the expression profile of miRNAs in Aedes albopictus / 南方医科大学学报

<p><b>OBJECTIVE</b>To verify the miRNA in Aedes albopictus and characterize the expression profile of several miRNAs across all the life stages of Aedes albopictus.</p><p><b>METHODS</b>Based on the published miRNA sequences of Anopheles gambiae and Aedes aegypti, 6 DIG-labeled antisense probes were synthesized. The total RNAs from Aedes albopictus in 6 developmental stages (embryo, early larvae, late larvae, pupa, male and female adults) were extracted with a mirVana miRNA isolation kit, loaded onto 15% denaturing polyacrylamide gel and hybridized with the appropriate DIG-labeled probes.</p><p><b>RESULTS</b>Northern blotting detected 5 miRNAs in Aedes albopictus, of which mir-9a was mainly expressed in embryo and larva stages, let-7 in pupa and adult stages, miR-184 in all life stages, mir-M1 only in the embryos and miR-1175 in all the life stages except for embryos. The expression profiles of these miRNAs in Aedes albopictus were similar to those in D. melanogaster and An.stepheni. miR-1174 was not detected in any of the developmental stages of Aedes albopictus.</p><p><b>CONCLUSION</b>These results present the first direct experimental evidence of miRNA in Aedes albopictus. The expression profiles of the analyzed miRNAs in Aedes albopictus showed stage specificity and conservation with other mosquitoes. Further studies on the functions of these miRNAs may offer new insights in mosquito biology and may lead to novel approaches to the development of insecticides.</p>

Bioinformatics analysis of mosquito densovirus nostructure protein NS1 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze and predict the structure and function of mosquito densovirus (MDV) nostructual protein1 (NS1).</p><p><b>METHODS</b>Using different bioinformatics software, the EXPASY pmtparam tool, ClustalX1.83, Bioedit, MEGA3.1, ScanProsite, and Motifscan, respectively to comparatively analyze and predict the physic-chemical parameters, homology, evolutionary relation, secondary structure and main functional motifs of NS1.</p><p><b>RESULTS</b>MDV NS1 protein was a unstable hydrophilic protein and the amino acid sequence was highly conserved which had a relatively closer evolutionary distance with infectious hypodermal and hematopoietic necrosis virus (IHHNV). MDV NS1 has a specific domain of superfamily 3 helicase of small DNA viruses. This domain contains the NTP-binding region with a metal ion-dependent ATPase activity. A virus replication roller rolling-circle replication(RCR) initiation domain was found near the N terminal of this protein. This protien has the biological function of single stranded incision enzyme.</p><p><b>CONCLUSION</b>The bioinformatics prediction results suggest that MDV NS1 protein plays a key role in viral replication, packaging, and the other stages of viral life.</p>

Electroacupuncture alleviates complete Freund's adjuvant peripheral chronic inflammatory pain in mice / 中国针灸

<p><b>OBJECTIVE</b>To probe into the law of acupuncture analgesic effect and specificities of acupoint action.</p><p><b>METHODS</b>Adult male Kunming mice were randomly divided into a control group, a model group, a model plus electroacupuncture (EA) group and a model plus sham EA group. Chronic inflammatory pain model was prepared by injection of complete Freund's adjuvant (CFA) into right posterior foot, and paw withdrawing latency (PWL) induced by radiation heat was used as pain threshold index, and changes of PWL in all the groups were investigated.</p><p><b>RESULTS</b>After modeling, PWL significantly shortened on the inflammatory side (P < 0.05); EA at bilateral "Zusanli" (ST 36) and "Kunlun" (BL 60) could significantly reverse the shortened PWL (P < 0.05), and this effect was prolonged along with increase of EA times, but in the sham EA group PWL did not significantly change; EA at bilateral "Shousanli" (LI 10) and Neiguan" (PC 6) could not reverse the shortened PWL (P > 0.05).</p><p><b>CONCLUSION</b>Analgesic effect of EA is strengthened along with increase of EA times and shows specificity of acupoints to a certain extent.</p>

Construction of the life cycle of Angiostrongylus cantonensis in laboratory / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the life cycle of Angiostrongylus cantonensis (A.cantonensis) in laboratory condition.</p><p><b>METHODS</b>SD rats were infected orally with the third-stage larvae of A.cantonensis collected from Jiangmen, Guangdong province. Six weeks after infection, the first-stage larvae were isolated from fresh feces of the rats by using Baermann funnel to infect 25 second-generation white jade snails raised in laboratory at the daily dose of 300 000 for 3 consecutive days. Three weeks later, the snails were dissected for counting the third-staged larvae of A.cantonensis, and those positive for A.cantonensis infection were fed directly to 10 fasting rats. The serum samples of the rats were then collected 2 weeks later for examination of specific antibodies using ELISA. The feces of the infected rats were examined microscopically after 6 weeks, and the brain, heart and lungs of the infected rats were dissected to observe the larvae at 3, 5, and 8 weeks, respectively.</p><p><b>RESULTS</b>The 3-stage larvae of A.cantonensis were found in the second-generation snails 3 weeks after infection. The positivity rate of serum specific antibodies was 100% in the 10 rats 2 weeks after feeding of the infected snails. The 1-stage larvae were detected in the feces of the rats 6 weeks after infection, and the fourth-stage larvae were found in the brain of the rats at 3 weeks, while adult worm and eggs were found in the heart and lungs of the infected rats at 5 and 8 weeks.</p><p><b>CONCLUSION</b>The successful establishment of human colon carcinoma cell line with PRL-3 gene knock-down provide a basis for investigation of the role of PRL-3 gene in the metastasis of human colorectal carcinoma.</p>

Screening and identification of therapeutic effect evaluation antigens of angiostrongyliasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To identify antigens which may help evaluate the therapeutic effect of angiostrongyliasis from adult worm antigen of Angiostrongylus cantonensis.</p><p><b>METHODS</b>The adult worm antigens of A. cantonensis were analyzed by Western blotting with the sera of rats infected with A. cantonensis before and after treatment. The sera of rats were tested by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The antigens with relative molecular mass between 38,000 and 78,000 reacted not only with the sera of rats before treatment, but also with that after treatment. The antigens with M(r) between 190,000 and 17,000 reacted with the sera of rats before treatment but not with that after treatment; those with M(r) between 32,000 and 24,000 antigens strongly reacted with the former, but the reaction became much weakened with the latter. The AC32-IgG antibody appeared earlier than the AC-IgG, and disappeared rapidly after treatment. Six of the 10 treated rats became negative for AC-IgG as found by ELISA.</p><p><b>CONCLUSION</b>The antigens of adult worm antigen of A. cantonensis with M(r) of 190,000, 32,000, 24,000, 17,000 and 16,000 may serve as candidate antigens for therapeutic effect evaluation of angiostrongyliasis.</p>

Familial pedigree analysis of Shen-yin deficiency syndrome in families with type 2 diabetes mellitus GU / 中国中西医结合杂志

To analyse the familial aggregation and genetic predisposition of Shen-yin deficiency syndrome (SYDS) in families with diabetes mellitus type 2 (DM2). Methods One hundred and forty-one DM2 patients were collected from 32 family lines in Nanjin area, in which the probands were differentiated as DM2 with SYDS. On them, genetic analysis on the characteristics of SYDS was conducted using pedigree analysis, morbidity and heritability of the first-degree relatives of the probands were calculated, and the action of familial SYDS factor on the genesis of the syndrome was assessed by multiple factors regression analysis. Results The morbidity rate of SYDS in the first-degree relatives of the probands was 33.71%, and the heritability, calculated by Falconer formula, was 80.6%. The fitting result of regression analysis showed that familial factor played an important role in SYDS genesis (OR = 5.61, P = 0.001), but DM2 itself is not an independent risk factor for it. Conclusion DM2 with SYDS shows the tendency of familial aggregation and genetic predisposition, genetic factor is associated with the genesis of the syndrome. Pedigree research is a good method for exploring the relationship between syndrome and genetic factor.