BCG Vaccine ; adverse effects ; Humans ; Immunologic Deficiency Syndromes ; complications ; Panniculitis ; etiology ; T-Lymphocytes ; Tuberculosis ; etiology

Aim To construct eukaryotic expression vector of shRNA(small hairpin RNA)for human SREBP-1(sterol regulation element binding protein-1)gene and explore its effects on lipid droplet formation in human renal proximal tubular epithelial cell line(HKC)under the stimulation of high glucose.Methods Two eukaryotic expression vectors of shRNA were constructed for human SREBP-1 gene.The HKC cells were transfected with negative control plasmid(pGenesil-1-HK)and two recombinant vectors(pGenesil-1-SREBP1-1 and pGenesil-1-SREBP1-2)and then were cultured under the stimulation of high glucose for about 48 h.The expression of SREBP-1 mRNA and FAS mRNA was detected by RT-PCR and SREBP-1 protein expression was investigated by Western blot.Lipid droplets were detected by Oil Red O staining.Results DNA sequencing showed that the target segments were successfully cloned into pGenesil-1 vector respectively.RT-PCR indicated that two recombinant vectors could inhibit the expression of SREBP-1 mRNA and FAS mRNA in HKC cells under the stimulation of high glucose.Similarly,SREBP-1 protein was also inhibited by the transfection with recombinant vectors.Oil Red O staining found that silencing of SREBP-1 gene resulted in lipid droplets decrease.Conclusions The eukaryotic expression vector of shRNA for human SREBP-1 gene was successfully constructed,and the expression of SREBP-1 was inhibited effectively by the expressed siRNA in HKC cells that resulted in lipid droplets decrease through FAS mRNA transcription inhibition.

A 69-year-old man presented with a 3-year history of scattered erythematous patches, perifollicular papules, acneiform lesions(such as milia, cysts) on the head, trunk and limbs as well as alopecia and peripheral eosinophilia. Histopathological examination revealed chronic focal dermal and perifollicular inflammatory infiltration with vascular proliferation and presence of a small number of eosinophils. He was initially diagnosed with folliculitis, and treated with antihistamines and antibiotics. Thereafter, lesional inflammation and pruritus were attenuated. However, plaques and alopecia developed in the occipital area 3 months later. The second histopathology of biopsy specimens revealed a dense dermal infiltrate of lymphoid cells and eosinophils, perifollicular infiltrate with numerous lymphoid cells, eosinophils and atypical lymphocytes migrating into hair follicles. Alcian blue stain showed epidermal mucinosis in follicles. Immunohistochemical examination showed positive staining of atypical cells for CD3, CD4, CD5, CD2, CD43 and ubiquitin carboxyl-terminal esterase L1 (UCHL1), but negative staining for CD20, CD79a, Epstein-barr virus, CD56, phosphoglucomutase-1, myeloperoxidase, CD7, or AE1 and AE3 monoclonal anti-keratin antibodies. T-cell receptor gene rearrangement was undetected. He was diagnosed with folliculotropic mycosis fungoides. Novel skin lesions still emerged after treatment with photochemotherapy (PUVA) plus acitretin. Follow up of the patient still continued.

Objective To provide reliable technical method by identifying referential anatomic landmarks for retroperitoneal laparoscopic renal surgery,with respect to the renal hilum and renal artery.Methods The regional anatomy of the posterior abdominal wall was studied in 35 cases of retroperitoneal laparoscopic renal surgery from January to August 2010.These included 27 cases of renal cancer,6 cases of renal pelvis cancer and 2 cases of renal tuberculosis.Distended the retroperitoneal space using balloon dilation along with sharp and dull dissection.We recorded the forms and positions of the posterior abdominal cavity's anatomical landmarks and evaluated the relationship between each anatomical landmark with respect to the renal hilum and renal artery.Results The perirenal fascia posterior layer and perinephric fat on the renal side were observed,and several anatomical landmarks gradually appeared on the posterior abdominal wall.The diaphragm extended across the upper retroperitoneal space near the superior pole of the kidney,and the psoas major and the quadratus lumborum muscles were located at the lower retroperitoneal space,near the inferior part of the kidney.The intersection of the upper diaphragm muscle with the lower psoas major and quadratus lumborum muscles were bordered by the lateral and medial arcuate ligaments.The lateral arcuate ligament arched across the upper part of quadratus lumborum,while the medial arcuate ligament arched across the upper part of psoas major.The medial arcuate ligament points extended towards the upper border of the renal hilum.These landmarks enable us to locate the position of the kidney,reach the renal hilum and identify the renal vessels in all 35 cases.Conclusions The relative position of the muscles and ligaments of the posterior abdominal wall are consistent and can be clearly seen under retroperitoneoscopy.Based on the position of the diaphragm and psoas major,the kidney can be located.In addition,based on the position of the medial arcuate ligament,the renal hilum and renal artery can be located.Assistance from these anatomical landmarks will simplify the retroperitoneal laparoscopic renal surgery.

Fifty-eight samples belonging to 7 species of Arisaematis Rhizoma and its adulterants were collected. The ITS2 locus was employed as a DNA barcode and amplified, sequenced and assembled for all of the collected samples. Then, ITS2 sequences have been annotated using HMM-based method. The intra- and inter-specific variations were calculated and NJ tree was constructed using MEGA 6.0 software. The results showed that inter-specific K2P distances were significantly larger than intra-specific distances for all of the three origin species of Arisaematis Rhizoma. Furthermore, three origin species, Arisaema amurense, A. erubescens and A. heterophyllum, can be respectively formed to be a single branch with high bootstrap values. It is concluded that ITS2 can be used to correctly identify Arisaematis Rhizoma from its adulterants and the application of ITS2 in the identification of traditional Chinese medicine has an important prospective.
Arisaema ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

Animals ; Inflammation ; chemically induced ; Lung ; drug effects ; Oligodeoxyribonucleotides ; pharmacology ; Pulmonary Alveoli ; drug effects ; pathology ; Silicon Dioxide ; adverse effects

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the relationship between DNA repair gene XRCC1 and XPD polymorphisms and individual susceptibility to prostate cancer. Methods PCR-restriction fragment length polymorphism assay was used to analyze the XRCC1 (C26304T and G28152A) and XPD A35931C polymorphisms in 358 prostate cancer patients and 312 healthy controls. Unconditional logistic regression analysis was performed to calculate odd ratio (OR) and 95% confidence interval (CD for estimating the correlation between different genotypes and prostate cancer risks. Results Forty-seven(13.1%) cases present XRCC1 28152AA genotype in prostate cancer group, while 24 cases in the control group (7. 1%), individuals with this genotype had a significantly increased risk for prostate cancer (OR 1. 924, 95%CI=1.126 - 3. 288, P=0. 017). There was no significant difference between two groups at XRCC1 C26304T and XPD A35931C sites. Combined analysis of the three sites polymorphisms showed that individuals with XRCC1 28152 AA and XPD 35931AC+CC genotype had a higher risk of prostate cancer than those with three wild genotypes (OR = 3. 087,95%CI 1. 081 - 8.813;OR = 3. 376,95%CI 1.067-10.683;OR 3. 216,95%CI=1. 439-7.188,P = 0. 004). Analysis stratified by age of onset, PSA, Gleason score and T stage revealed that XRCC1 28152AA and XPD 35931 AC+CC high-risk genotype was especially associated with early age at onset of prostate cancer (P<0. 05). Conclusions The XRCC1 and XPD genotypes may be contributed to the risk of developing prostate cancer, particularly for younger patients.

Aim To investigate the effects of suppressor of cytokine signaling-1(SOCS-1)on advanced glycation end products(AGEs)induced-renal tubular epithelial-myofibroblast transdifferentiation and activation of JAK/STAT in cultured human renal tubular epithelial cells(HKC).Methods Stable transfections of HKC with pCR3.1 vector and pCR3.1/SOCS-1 were performed with Lipofectamine 2000,and cells were selected with geneticin.Cells were stimulated with BSA and AGEs. The protein expressions of SOCS-1,α-SMA,E-cadherin,Col I,signal transducer and activatior of transcription 1,3(STAT1,STAT3),p-STAT1 and p-STAT3 were observed by Western blot.The protein synthesis of TGF-β_1 in the supernatants of the HKC was detected by enzyme-linked immunoadsorbent assay(ELISA).α-SMA and E-cadherin mRNA were measured by reverse transcription and polymerase chain reaction(RT-PCR).Results Compared with control group,the expression levels of α-SMA protein and mRNA and Col I were significantly increased in HKC with AGEs stimulation and there was a higher concentrations of TGF-β_1 in the supernatants.However,the expression of E-cadherin protein and mRNA were decreased with AGEs stimulation.Overexpression of SOCS-1 inhibited AGEs-induced activation of STAT1 and STAT3 and high expression of α-SMA protein and mRNA and Col I,and reversed the expression of E-cadherin protein and mRNA induced by AGEs.Meanwhile,overexpression of SOCS-1 reduced the concentration of TGF-β_1 in the supernatants of HKC with AGEs stimulation.Conclusion Overexpression of SOCS-1 inhibits AGEs-induced renal tubular epithelial-myofibroblast transdifferentiation maybe partly through blocking activation of JAK/STAT.

Objective To investigate the effect of suppressor of cytokine signaling-1 (SOCS-1)on expression of monocyte chemoattractant protein-1 (MCP-1)in human glomerular mesangial cells (HMCs) under high concentration of glucose. Methods Stable transfections of HMC with pCR3.1 vector and pCR3. 1-SOCS-1 were performed with hpofectamine 2000, and cells were selected with geneticin. Cells were stimulated with low glucose (LG, 5.5 mmol/L), high glucose (HG, 30 mmol/L), LG plus mannitol (24.5 mmol/L) and AG490 (10 μmol/L). The protein expression levels of SOCS-1, signal transducer and activators of transcription 1,3 (STAT1, STAT3),p-STAT1 and p-STAT3 were observed by Western blotting. The protein synthesis of MCP-1, FN and type Ⅳ collagen in the supernatants of the HMCs were detected by ELISA and radioimmunoassay. The expression level of SOCS-1 and MCP-1 mRNA was measured by BT-PCR.Results HG induced the expression of SOCS-1 protein and mRNA in HMCs in time-dependant manner, peaked at 4 h, and gradually decreased to baseline at 24 h. Compared with low glucose control group, the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.39±0.05) vs (0.16±0.02)]were significantly increased in HMCs under high glucose medium (P ＜0.01 ). Exposure of HMCs to high glucose conditions showed high concentration of MCP-1 [(459±67) ng/L vs (241±19) ng/L], fibronectin [(5.84±0.61) mg/L vs (3.41±0.31) mg/L]and type Ⅳ collagen [(16.45±2.30) μg/L vs (9.56±1.52) μg/L]in the supernatants (all P＜0.01).Overexpression of SOCS-1 inhibited the phosphorylation levels of STAT1 and STAT3 and the expression of MCP-1 mRNA [(0.34±0.04) vs (0.42±0.05)]in HMCs under high glucose condition (all P＜0.05). Compared with vector control group, the concentration of MCP-1 [(387±47) ng/L vs (463±56) ng/L], fibronectin [(4.61±0.57) mg/L vs (5.76±0.74) mg/L]and type Ⅳ collagen [(13.4±2.32) μg/L vs (17.1±2.57) μg/L]was decreased in supernatants of HMCs with SOCS-1 overexpression (all P＜0.05). Compared with HG group, the expression of MCP-1 mRNA (0.31±0.04) and the concentration of MCP-1 [(361±53) ng/L], FN [(5.46±0.71) mg/L]and type Ⅳ collagen [(15.2±1.97) μg/L]in supernatants were decreased in HMCs treated with AG490.Conclusion Overexpression of SOCS-1 inhibits overproduction of MCP-1 and ECM proteins in HMCs under high glucose conditions, which may be partly by regulating the phosphoryhtion of STAT1 and STAT3.