AIM: To investigate the correlation of hepatitis B virus X protein (HBx) with renal tubular epithelialcell apoptosis in hepatitis B virus-associated glomerulonephritis (HBVGN) and the possible signaling mechanism. METHODS: The activation of JAK2/STAT3 signal pathway and the expression of apoptosis -related proteins in humankindey proximal tubular epithelial cells (HK-2 cells) were determined by Western blotting after transfection with HBx eukaryoticexpression vector.The cell proliferation was observed by CCK-8 assay.The cell apoptosis was analyzed by the imagingof HO33342 staining, transmission electron microscopy and flow cytometry with Annexin V /PI double staining.RESULTS:After transfection of the target gene HBx, the expression levels of both p-JAK2 and p-STAT3 were significantly increased.At the same time, the cell proliferation was obviously inhibited, and the apoptotic rate was increased.After incubationwith AG490, the JAK2/STAT3 signal pathway was partially blocked, and the cell apoptosis induced by HBx was reduced. CONCLUSION: HBx up-regulates the activation of JAK2/STAT3 signal pathway to induce renal tubular epithelialcell apoptosis, which is possibly involved in the pathogenic mechanism that HBV directly damages nephridial tissue .

[ABSTRACT]AIM:ToinvestigatetheeffectofmiR-155-specificsiRNAaloneorincombinationwithcytosinear-abinoside (Ara-C) on the growth and apoptosis of Burkitt lymphoma Raji cells .METHODS: miR-155-specific siRNA and/or Ara-C were used to treat the cells .Quantitative real-time polymerase chain reaction was used to detect the expres-sion of miR-155.The growth of the cells was analyzed by CKK-8 assay.The cell apoptosis was determined by flow cytome-try.RESULTS:The miR-155 expression level of the cells transfected with miR-155 siRNA was significantly lower than that in the 2 control groups .Ara-C or miR-155 siRNA alone inhibited the growth of Raji cells in a dose-depend manner . miR-155 siRNA combined with Ara-C produced more inhibition of cell proliferation (P<0.05).After treatment for 48 h, the apoptotic rate of Raji cells in miR-155 siRNA+Ara-C group [(38.4 ±1.4)%] was higher than that in Ara-C group [(16.5 ±0.3)%] and miR-155 siRNA group [(14.6 ±0.3)%], with statistically significant difference (P<0.05). The expression of caspase-3 in Ara-C+miR-155 siRNA group was increased significantly as compared with Ara-C group and miR-155 siRNA group.CONCLUSION:miR-155-specific siRNA enhances the chemosensitivity of Raji cells to Ara-C by inducing apoptosis through the caspase-3 pathway .

[ABSTRACT]AIM:Toinvestigatecellapoptosisindiabeticfootulcersandtheeffectofadvancedglycosylation end products (AGEs) on apoptosis in human fibroblast cells.METHODS: Diabetic foot patients (n=18) and 18 age-matched non-diabetic controls were recruited .The clinical and biochemical features were compared by statistics methods . Skin biopsies were obtained from foot .Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex ( SABC ) staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling ( TUNEL) technique was used to detect apoptosis of the skin tissues .Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose ( changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h) or AGEs (150 mg/L, containing 5.6 mmo/L glucose).After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3.Other cells were trypsinized , washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis .RE-SULTS:Diabetic patients had higher levels of fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin A1c (HbA1c), and longer wound duration.The protein level of cleaved caspase-3 was signifi-cantly higher in diabetic group , suggesting that apoptosis was increased in diabetic skin tissues .TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group (8.4%±1.5% vs 3.8%± 0.8%) , which further confirmed that cell apoptosis was increased in diabetic foot tissues .In human fibroblasts , the levels of cleaved caspase-3 in normal group , sustained high glucose group , fluctuant high glucose group and AGEs group were 0.80 ±0.13, 1.22 ±0.18, 1.46 ±0.32 and 1.83 ±0.25, respectively.The apoptotic rates detected by flow cytometry were 2.43%±0.19%, 2.89%±0.51%, 3.99% ±0.24% and 6.83% ±0.36%, respectively.Both the level of cleaved caspase-3 and the apoptotic rate in AGEs group were higher than those in normal glucose group and sustained high glucose group .CONCLUSION:Increased apoptosis in diabetic foot ulcers is one of the most important reasons for im-paired wound healing .As compared to sustained high glucose and glucose fluctuations , AGEs induce greater apoptosis in human fibroblast cells .

AIM: To explore the effect of hydrogen sulfide on the senescence of human umbilical vein endothe -lial cells (HUVECs) induced by high glucose.METHODS: Senescence model was established by treating HUVECs with33 mmol/L glucose for 48 h.The parameters were detected to demonstrate the effect of hydrogen sulfide on senescence andthe mechanism involved was also investigated .RESULTS: In the cells treated with high glucose, the proliferation was attenuatedwith a higher number of senescence -associated β-galactosidase (SA-β-Gal) positive cells, and plasminogen activatorinhibitor 1 (PAI-1) protein expression, malondialdehyde (MDA) production and NF-κB p65 activity were increasedsignificantly, but the expression of superoxide dismutase 1 (SOD1) was decreased.However, the cell number and SOD1expression were increased, and the number of SA-β-Gal positive cells, PAI-1 protein expression, MDA production and theactivity of NF-κB p65 were decreased after sodium hydrosulfide (100 and 200 μmol/L) treatment.CONCLUSION: Exogenoushydrogen sulfide prevents HUVECs against high glucose -induced senescence by suppressing oxidative stress and NF -κB p65 activity.

[ABSTRACT]AIM:Toestablishatransgenicheterozygousmousemodelofprecancerouslesionsofcolorectal cancer with p110δmutation in the C57BL/6J background for serving the studies on colorectal cancer research mediated by p110δ.METHODS:The transgenic heterozygous mice were generated by crossing in p110δD910A/D910A mouse and ApcMin/+mouse, and the genotype was detected by PCR .Compared with ApcMin/+mice, transgenic heterozygous mice ( ApcMin/+;p110δD910A/D910A)were counted, and the number and size of intestine polyps were analyzed after methylene blue staining . The intestinal tissue structure was assessed by HE staining .RESULTS:The transgenic heterozygous mouse model of pre-cancerous lesions of colorectal cancer with p 110δmutation was established .The number and size of polyps in the transgenic heterozygous mice were declined .CONCLUSION: A transgenic heterozygous mouse model of precancerous lesions of colorectal cancer with p 110δmutation was successfully established .The initial phenotype of intestinal tumors in transgenic mice was observed .This model will greatly contribute to the related research of colorectal cancer in mice .

[ABSTRACT]AIM:ToestablishandevaluateahyperbilirubinemiaandkernicterusmodelinneonatalSDrats. METHODS:Three-day-old SD rats were randomly divided to 7 experimental groups by litter and body weight , and were in-traperitoneally injected with physiological saline (control group), and 6.25μg/g (T1), 12.5μg/g (T2), 25μg/g (T3), 50μg/g (T4), 100μg/g (T5) and 200μg/g (T6) bilirubin, respectively, twice every day for 3 d.All rats were photo-graphed , weighed and killed 12 h after the last injection .The contents of the stomach were drawn and weighed , and the index was calculated .The liver/body weight ratio was determined , the total and unconjugated bilirubin in the serum and total bili-rubin in the brain were calculated , and the contents of ATP and water in the brain were measured .HE and Nissl staining were used to observe the pathological changes .RESULTS:Along with the increase in bilirubin , gradual exacerbation of the general performance of the rats , and yellowish discoloration of the skin and mucous membranes were observed .The degree of the activity gradually reduced , and the weight gain was suppressed .The weight of T6 group showed negative growth , and the 72 h mortality rate was close to 100%.The mortality rate in T4 and T5 groups continued to rise 1 week after injection .Com-pared with control group , the weight of stomach contents and stomach content index in T 3~T5 groups significantly decreased (P<0.01).The liver/body weight ratio in T5 group was significantly higher (P<0.05).The concentrations of serum total and unconjugated bilirubin and brain bilirubin levels in T 1~T5 groups were gradually increased , while the brain water con-tent had no difference among groups .The brain ATP content in T1~T5 groups increased at the beginning and reached its peak in T3 group, but compared with control group , that in T4 group and T5 group significantly reduced (P<0.05).HE re-sults showed that , with the increase in bilirubin concentration , the number of the neurons in the cerebral cortex of the rats de-creased.In T4 group and T5 group, the neuronal structural disorder , cell swelling, nuclear pyknosis, fragmentation and dis-solution, increase in non-homogeneous structure of the material dyed red , and disappearance of nuclear staining were ob-served.Nissl staining showed that , compared with control group , in T1 group and T2 group, the cortical neurons became smaller, Nissl bodies decreased , and cytoplasmic staining changed little .The cortical neuronal tigroid body color became light gradually, neuron cells become small , and Nissl bodies decreased obviously in T 3, T4 and T5 groups.The T4 and T5 rat ce-rebral cortical neurons dissolved or even disappeared .CONCLUSION:Newborn 3-day-old SD rats receiving intraperitoneal injection of bilirubin at doses of 12.5, 25, 50 and 100μg/g, 2 times a day, can induce hyperbilirubinemia , and 50 and 100μg/g can cause bilirubin encephalopathy .

[ABSTRACT]Pulmonaryarterialhypertensionisdefinedasamultifactorialgroupofpulmonaryvasculardisorders characterized by a progressive increase in the pulmonary vascular resistance , resulting in right heart failure and premature death.The plexiform lesion is the hallmark of severe pulmonary arterial hypertension .This article summarized the recent progress in the plexiform lesion including its occurrence , structure, animal models and molecular mechanism , which tried to predict the tendency of plexiform lesion study .

[ABSTRACT]IL-13isapleiotropiccytokinemainlysecretedbyactivatedTh2cells.Ithas2receptors,IL-13Rα1 and IL-13Rα2.The latter had been thought to serve exclusively as a decoy receptor for a long period of time due to its short cytoplasmic tail and lack of signal transduction structure .Since Fichtner-Feigl reported in Nature Medicine that IL-13 is in-volved in induction of TGF-βproduction and tissue fibrosis through IL-13Rα2-mediated signaling pathway , it was found that IL-13Rα2 has more sophisticated functions than just a simple decoy receptor as more and more researches have explored its signaling functions .This review combines the most advanced research results with previous investigation and discusses the gene structure, expression, production, distribution, subtype conversion and possible signal pathways mediated by this re-ceptor.More importantly, the connection with human diseases and the applications in disease diagnosis and molecule targe -ted therapy for cancer are also discussed .

[ABSTRACT]AIM:ToinvestigatetheeffectoforidoninontheinvasionandmigrationofhumanlungcancerNCI-H460 cells.METHODS:NCI-H460 cells were divided into high-dose (HD), middle-dose (MD) and low-dose (LD) oridonin groups (cultured with 40, 20 and 10μmol/L of oridonin, respectively, as experimental groups), and normal (N) group ( treated without oridonin as control ) .The cell growth was observed .The cell proliferation was detected by MTT as-say.Boyden chamber was used to determine the cell invasive capacity .The cell migration was also measured .The levels of MMP-2 and MMP-9 were assayed by Western blotting .RESULTS:The cell counts in the experimental groups were lower than that in N group .The cell proliferation was inhibited as the inhibitory rates were 48.94%, 36.17%and 19.15% for HD group, MD group and LD group, respectively.The numbers of the invasive cells were 26.67 ±5.16 for HD group, 36.17 ±5.08 for MD group, and 44.33 ±5.50 for LD group.The migration rates in the experimental groups were lower than that in N group .The expression of MMP-2 and MMP-9 decreased dependent on the oridonin dose as follows: HD group

[ABSTRACT]AIM:Toinvestigatetheroleofcysteine-rich61(Cyr61/CNN1)inproliferationandmigrationof bone marrow mesenchymal stem cells ( BMSCs ) .METHODS: The lentiviral vector carrying CCN 1 ( Lenti-GFP-CCN1 ) was constructed and then transfected into the rat BMSCs .The cells were divided into non-transfection group , transfection group ( transfected with Lenti-GFP-CCN1 ) and negative control group ( Lenti-GFP ) .The fluorescence intensity of the transfected BMSCs was observed under inverted fluorescence microscope .The effects of CCN1 on the proliferation and mi-gration of BMSCs were detected by MTT assay and scratch wound healing assay .RESULTS:The proliferation of BMSCs transfected with Lenti-GFP CCN1 had no significant difference compared with negative control group and control group .The width/thickness ratio of migrated BMSCs in wound healing was significantly higher in Lenti-GFP-CCN1 group than that in negative control group and control group (P<0.05).CONCLUSION:Exogenous CCN1 promotes the migration of BMSCs.