AIM: To evaluate the anatomic and visual outcomes of 25-gauge vitrectomies combined with air tamponade for the treatment of idiopathic macular hole (IMH).METHODS: Thirty eyes of 27 patients with IMH were included in this prospective interventional study.All patients underwent 25-gauge pars plana vitrectomy (PPV) combined with phacoemulsification and air tamponade.Best corrected visual acuity (BCVA, logMAR), perimetry and multifocal electroretinography (mfERG) were conducted before and after the operation.Anatomical changes were evaluated with optical coherence tomography (OCT).RESULTS: The macular holes closed successfully in 28 eyes after the primary vitrectomy.The mean BCVA improved from 0.72±0.22 logMAR preoperatively to 0.29±0.18 logMAR postoperatively (P<0.001).In the visual field of central 10°, the average mean deviation (MD) decreased from-3.59±1.83 dB preoperatively to-2.51±1.36 dB postoperatively (P<0.001) and the average pattern standard deviation (PSD) decreased from 1.86±0.68 dB preoperatively to 1.33±0.32 dB postoperatively (P=0.001).The retinal response densities of mfERG in the foveal and perifoveal area increased significantly, and implicit times of rings 4-6 prolonged significantly (P<0.05).The symptom duration and baseline N1 amplitude densities at ring 1 had a significant impact on postoperative BCVA (P<0.001, P=0.001, respectively).CONCLUSION: The 25-gauge PPV and air tamponade with 1 day prone positioning produce favorable anatomic and functional outcomes.

To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B ; genetics ; metabolism ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Caco-2 Cells ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Up-Regulation ; drug effects

OBJECTIVETo introduce the WHO 2000 diagnostic criteria of biopsy of colorectal intraepithelial neoplasia and carcinoma and to enhance diagnostic accuracy and avoid overdiagnosis and underdiagnosis.

METHODThe postoperative pathological examination and preoperative biopsy in 56 patients diagnosed as colorectal intraepithelial neoplasia and carcinoma before operation from January 2001 to October 2005 were compared retrospectively.

RESULTSAmong the 56 cases, 16 patients were diagnosed by preoperative biopsy as carcinoma in situ, intramucosal carcinoma and adenocarcinoma, but according to the new standard, of them 14 cases should be revised to be higher grade colorectal intraepithelial neoplasia.

CONCLUSIONSStrictly adhere to the new WHO criteria, colorectal intraepithelial neoplasia and carcinoma can be diagnosed properly, but for the cases that submucosal muscular layer would not presented in biopsy, the diagnosis should be made by combining clinical findings and various examination results so as to avoid underdiagnosis and delay of treatment.

Adult ; Aged ; Biopsy ; Carcinoma ; diagnosis ; pathology ; Carcinoma in Situ ; diagnosis ; pathology ; Colon ; pathology ; Colorectal Neoplasms ; diagnosis ; pathology ; Diagnosis, Differential ; Female ; Humans ; Male ; Middle Aged ; Preoperative Care ; Rectum ; pathology ; Retrospective Studies

OBJECTIVETo investigate the expression and the localization of Cathepsin K and IL-6 mRNA in root-resorbing tissue and to elucidate the molecular changes and mechanism of root resorption induced by tooth movement.

METHODSRats were subject to experimental tooth movement to induce root resorption. In situ hybridization was performed to identify the cells in root-resorbing tissue that produced Cathepsin K or IL-6 the difference of CK mRNA or IL-6 mRNA expression between root resorption group and control group was calculated by t-test.

RESULTSCathepsin K mRNA was highly and selectively expressed in multinuclear odontoclast and IL-6 mRNA expressed in fibroblast, osteoblast, osteocyte and cementoblast. The expression of Cathepsin K mRNA and IL-6 mRNA in root-resorbing tissue increased evidently compared with the normal periodontium.

CONCLUSIONSOdontoclast in the root-resorbing tissue expresses Cathepsin K mRNA that participates in proteolysis during root resorption. IL-6 plays a very important role in the root resorption as a multifunctional cytokine.

Animals ; Cathepsin K ; Cathepsins ; biosynthesis ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Male ; Osteoclasts ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Tooth Movement Techniques ; Tooth Resorption ; enzymology

OBJECTIVETo explore the cure effects of Jiangu Fufang on osteoporotic model induced by ovariectomy.

METHODRats were ovariectomized and administered drugs for 3 monthes. Bone mineral density and biomechanics properties, histomorphometric analysis and biochemical index such as calcium, phosphorus, alkaline phosphatase were detected.

RESULTJiangu Fufang could significantly increase bone density and biomechanics properties. The level of calcium, phosphorus and alkaline phosphatase were restored by Jiangu Fufang. Jiangu Fufang could significantly increase area of bone trabecula, thickness of cortical bone and bone trabecula.

CONCLUSIONJiangu Fufang could cure osteoporosis through increasing bone mineral density, improving bone biomechanics properties, and effecting bone metabolism.

Alkaline Phosphatase ; blood ; Animals ; Biomechanical Phenomena ; Bone Density ; drug effects ; Calcium ; blood ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; therapeutic use ; Female ; Femur ; physiopathology ; Lumbar Vertebrae ; drug effects ; pathology ; physiopathology ; Osteoporosis ; blood ; drug therapy ; physiopathology ; Ovariectomy ; Phosphorus ; blood ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To explore the demographic characteristics and clinical features of patients with idiopathic pulmonary arterial hypertension ( IPAH ) in China.Methods Between March 2007 and September 2010,IPAH diagnosis was confirmed by right heart catheterization in 150 adult patients from 31 clinical centers in China.Clinical and hemodynamic data were analyzed and patients were divided into WHO functional class Ⅰ / Ⅱ and WHO functional class Ⅲ/Ⅳ group.Results The mean age of 150 patients were 36 ± 13 years with female patient/male patient ratio of 2∶1,and mean BMI was ( 21.3 ± 3.5 ) kg/m2.Fatigue (n =123,82.0％ ) and dyspnea ( n =112,74.7％ ) are the most common symptoms.Accentuated pulmonic second sound ( P2 ) was detected in 92.0％ ( n =138 ) of patients during physical examination,which was also the most common sign.About 49.0％ ( n =73 ) patients were WHO functional class Ⅰ/ Ⅱ patients and 46.0％ ( n =68 ) patients were WHO functional class Ⅲ/Ⅳ patients. Six minutes walking distance (6MWD) and Borg dyspnea score was (337 ± 101 ) m and 2.0 (2.0,4.0),respectively.Right ventricular hypertrophy was suggested by ECG in 93.1％ ( n =140 ) patients.Right atrial pressure was (10 ±6) mm Hg,mean pulmonary artery pressure was (61 ±16) mm Hg,cardiac index was (2.3 ±0.8)L · min-1 · m-2 and pulmonary vascular resistance(1484 ±699) dyn · s-1 · cm-5 in this cohort.6 MWD (305 m ±89 m vs.377 m ±88 m) was significantly shorter while Borg dyspnea score [3.0 (3.0,5.0) vs.2.0 (2.0,3.0) ] was significantly higher in WHO functional class Ⅲ/Ⅳ patients than in WHO functional class Ⅰ/Ⅱ patients. Similarly hemodynamic parameters were also worse in WHO functional class Ⅲ/Ⅳ patients than in WHO functional class Ⅰ / Ⅱ patients ( all P ＜ 0.05).Conclusion Idiopathic pulmonary arterial hypertension patients in this cohort affect mostly young adults,dominated by female gender and lower body mass index. Fatigue and dyspnea are the most common symptoms and accentuated pulmonic second sound ( P2 ) is the most common sign.IPAH patients are often displaying severe functional and hemodynamic disturbance at first visit to hospitals.Dyspnea and hemodynamic impairment are related to 6MWD and WHO functional class.

BACKGROUNDHuman interleukin-10 (hIL-10) is a cytokine synthesis inhibitory factor, which is involved in various immune responses. The purpose of this study was to construct an adenoviral vector carrying the hIL-10 gene for expression of biologically active hIL-10 in rat bone marrow mesenchymal stem cells (rMSCs).

METHODSA pSNAV2.0-hIL10 plasmid was used as a template to obtain a hIL-10 cDNA fragment that was subcloned by restriction enzyme digestion and ligation into a pDC316-IRES-EGFP-lacZ alpha plasmid carrying an enhanced green fluorescent protein (EGFP) marker gene. The pDC316-hIL-10-IRES-EGFP plasmid was linearized by PmeI digestion and used to transfect HEK293 packaging cells using the adenovirus packaging system AdMax. Virus particles were amplified by repeatedly infecting HEK293 cells with the seed virus and then purified by ion exchange. After the number of virus particles and titer was determined, rMSCs were infected with the adenoviral vector. The infection rate was determined by fluorescence microscopy and flow cytometry, and hIL-10 protein expression in rMSCs was measured by Western blotting.

RESULTSThe virus particle concentration, OD260/280 value and virus titer of the amplified and purified recombinant adenovirus were 3.2 × 10(11) VP/ml, approximately 2.0, and 1.1 × 10(10) TCID50/ml, respectively. Bright green fluorescence was observed by fluorescence microscopy and flow cytometry in the recombinant adenovirus-infected rMSCs. GFP expression was considered the multiplicity of infection (MOI) and was time-dependent. The infection rate was 92.9% at 100 MOI.

CONCLUSIONSA bicistronic recombinant adenoviral vector for hIL-10 and EGFP gene expression were successfully constructed. The infection rate of rMSCs by the adenovirus was high (92.9% at 100 MOI) and the target gene hIL-10 was highly expressed in cells. The present study provides an experimental basis for further research of immunosuppressive therapy using hIL-10. The expression level of hIL-10 protein as detected by Western blotting was also MOI- and time-dependent.

Adenoviridae ; genetics ; Animals ; Cell Line ; Cells, Cultured ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Interleukin-10 ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; metabolism ; Rats

No abstract available.
Humans ; Interferon-gamma* ; Pityriasis Rosea*

OBJECTIVETo clone a novel human testis-specific gene TDRG1.

METHODSA new contig of expression sequence tags (ESTs) Hs.180197 was identified from the testis libraries using digital differential display (DDD) to screen the novel human testis-specific gene. To validate the use of bioinformatics approaches in gene discovery, the ESTs Hs.180197, which was predicted to be testis specific, was chosen for further study. Reverse transcriptase-polymerase chain reaction (RT-PCR) was performed on different normal tissues to identify the expression of Hs.180197 in human testis. Using bioinformatics methods and IMAGE cloning of this contig, the full-length cDNA sequence of the noval human gene was cloned.

RESULTSThis novel gene was 1197 bp in length, located in chromosome 6p21.1-p21.2. The sequence of the open reading frame was 504-806 bp, as confirmed by RT-PCR and sequencing in human testis. The cDNA encodes a novel protein of 100 amino acids with a theoretical molecular weight of 10 000 and isoelectric point of 6.81. The sequence shares no significant homology with any known protein in the databases. Semi-quantitative RT-PCR analysis of multiple tissues further showed that the novel gene was expressed specifically in adult human testis. Considering a possible relation of this novel gene with the function of human testis, we named this new gene TDRG1 (testis development related gene 1, GenBank accession number: DQ168992).

CONCLUSIONDDD combined with laboratory validation is an efficient method for identifying new human functional genes.

Adult ; Cloning, Molecular ; DNA, Complementary ; genetics ; Databases, Genetic ; Gene Expression Regulation ; Homeodomain Proteins ; genetics ; Humans ; Male ; Molecular Sequence Data ; Organ Specificity ; Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Testis ; metabolism

BACKGROUNDThe cannabinoid receptor-2 (CB2) is important for bone remodeling. In this study, we investigated the effects of CB2 selective antagonist (AM630) on receptor activator of nuclear factor kappa B (RANK) ligand (RANKL) induced osteoclast differentiation and the underlying signaling pathway using a monocyte-macrophage cell line-RAW264.7.

METHODSRAW264.7 was cultured with RANKL for 6 days and then treated with AM630 for 24 hours. Mature osteoclasts were measured by tartrate-resistant acid phosphatase (TRAP) staining using a commercial kit. Total ribonucleic acid (RNA) was isolated and real-time reverse transcriptase-polymerase chain reaction (RT-PCR) was done to examine the expression of RANK, cathepsin K (CPK) and nuclear factor kappa B (NF-κB). The extracellular signal-regulated kinase (ERK), phosphorylation of ERK (P-ERK) and NF-κB production were tested by Western blotting. The effect of AM630 on RAW264.7 viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay.

RESULTSAM630 did not affect the viability of RAW264.7. However, this CB2 selective antagonist markedly inhibited osteoclast formation and the inhibition rate was dose-dependent. The dose of ≥ 100 nmol/L could reduce TRAP positive cells to the levels that were significantly lower than the control. AM630 suppressed the expression of genes associated with osteoclast differentiation and activation, such as RANK and CPK. An analysis of a signaling pathway showed that AM630 inhibited the RANKL-induced activation of ERK, but not NF-κB.

CONCLUSIONAM630 could inhibit the osteoclastogenesis from RAW264.7 induced with RANKL.

Animals ; Blotting, Western ; Cell Differentiation ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Indoles ; pharmacology ; Mice ; Osteoclasts ; cytology ; drug effects ; metabolism ; RANK Ligand ; pharmacology ; Receptor, Cannabinoid, CB2 ; antagonists & inhibitors ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects