Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Intraoperative Complications ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Tracheotomy ; adverse effects ; methods ; Young Adult

BACKGROUND :Although the conventional 20 G pars plana vitrectomy can improve the prognosis of vitreous-retina diseases,but it would also cause serious operation related injury and much complications.OBJECTIVE:To evaluate the practicability and safety of 25-gauge transconjunctival sutureless vitrectomy system (TSV25G) in the macular surgery, so as to elicit the preliminary clinical experience of mimimally invasive vitrectomy system.DESIGN:Case analysisSETTING: Department of Ophthalmology, General Hospital of Chinese People's Armed Police Force; Department of Ophthalmology, Beijing Sino Japen Friendship Hospital; Department of Ophthalmology and Visual Science, Chinese University of Hong Kong; Department of Ophthalmology,Prince of Wales Hospital, Hong Kong PARTICIPANTS: From July 2003 to July 2004, 16 patients with macular diseases were admitted to the Department of Ophthalmology in the General Hospital of Chinese People's Armed Police Force and taken as subjects (n=16), they were confirmed as epiretinal membrane in 9 eyes, idiopathic macular hole in 3 eyes, traumatic macular hole in 2 eyes and vitreo-macu lar tractional syndrome in 2 eyes. The course of disease ranged from 3 to 36 months with eyesight varied between 0.06 and 0.4.METHODS: 16 eyes underwent preoperative ophthalmic evaluation in cluding visual acuity measurement, Amsler chart test, slit lamp microscopy,.indirect ophthalmoscopy tonometry and optical coherence tomography (OCT). For the macular hole patients OCT was performed. 25-gauge Transconjunctival Sutureless Vitrectomy System (TSV25G) was used to carry on minimally invasive surgery under anesthesia state, the surgical parameter was set as: high speed cutter with rate of 1500 cuts per minute,BSS bottle was hang at height of 40-50 cm, the maximum aspiration of TSV25G was 550 mm Hg with the intraocular pressure remained at around 29-35 mmHg during the operation. Three-beveled trocar were used to make three-transeonjunctival incision on sclera of about 0.5 mm long respectively in the infratemporal, superotemporal, and supraotemporal quan drants, meanwhile a transconjunctival cannula was placed. One end of the infusion tube was inserted into the infratemporal cannula to establish infusion with the other two cannulas used for the intraocular operation with 25-gauge vitreous cutter and other instruments, such as vitreoectomy and membrane dislocation. The surgery was terminated by removing the cannula except for ceaseless leak, the conjunctival and scleral incision were not sutured. Patients received follow-up examination for 1-12 months postoperatively.MAIN OUTCOME MEASURES:①The operative time and the time for the establishment of three access. ② Changes of intraocular pressure after operation. ③ The effusion from the puncture after the intraocular perfor mance. ④ The postoperative vision, operative complication and the sealing of the holes.RESULTS:All of the 16 patients completed the vitrectomy; and entered the data analysis.①16 patients complete the vitrectomy with the operative time of 28-56 minutes (the mean of 37 minutes). The average time for the establishment of three access and closing was 84 s and 32 s respectively.②The average preoperative intraocular pressure was 16.4 mm Hg,comparing with 13.5,15.5,17.9 mm Hg at postoperative 1 day, 1 w and 1 m. ③Water leakage were found in 4 wounds of three patients after operation, 3 wounds were sealed after injecting 1-2 mL disinfected air into the eyes, the other one sutured with 6-0 absorbable suture. ④The mean time of inpatients were 5 days postoperatively. The visual acuity improved in 14 patients by the average of three lines, amongst which visual acuity was found improved to above 0.8 in 5 patients and the visual distortion disappeared in 8 eyes and attenuated in 3 eyes. But still there were 2 cases without improvement.No surgical complications were noted and macular hole gained clinical sealing in totally 5 eyes.CONCLUSION: TSV25G system can simplify the surgical procedure with many other advantages such as decreasing the operation related injury and complications, as well as shortening operative time, which benefits the rapid rehabilitation.

This study investigated the role of Echinococcus multilocularis vesicle-derived extracellular vesicles(EVs)con-taining emu-miR-10-5p in regulating HSC activation in vitro.Mice were injected intraperitoneally with protoscoleces to con-struct a secondary alveolar echinococcosis model,and E.multilocularis vesicles were cultured in mice and identified by PCR.The EV structure in cultured vesicle tissue was observed by transmission electron microscopy.EVs were isolated from vesicle culture supernatants through differential-ultracentrifugation,and identified by transmission electron microscopy,nanoparticle size analysis,and western blotting.The activity of HSCs treated with different EV concentrations was detected with CCK-8 assays.The expression levels of α-SMA,and collagen Ⅰ and Ⅲ were detected by RT-qPCR and western blotting after EV treatment of HSCs.Bioinformatics analysis identified the specific miRNA emu-miR-10-5p.The proliferation of HSCs was detected with EdU after overexpression of emu-miR-10-5p,and the expression levels of α-SMA,collagen Ⅰ,and collagen Ⅲ in cells were detected by RT-qPCR and western blotting.Detection of α-SMA,collagen Ⅰ,and collagen Ⅲ expression levels in HSCs was performed after EVs were extracted from the culture supernatant of vesicular tissues with emu-miR-10-5p knockdown.E.multilocularis vesicles were successfully cultured from focal material in mice,and the E.multilocularis origin was confirmed through PCR amplification of specific genes.TEM observation of culture vesicles in-dicated abundant surface vesicular structures.E.multilocularis vesicle-derived EVs were isolated by differential-ultracentrifu-gation,and confirmed by TEM to be spherical and membrane like-structures.Vesicle-derived EVs were internalized by HSCs,and promoted the proliferation and activation of HSCs.Comprehensive analysis of miRNA sequencing results revealed emu-miR-10-5p expression in E.multilocularis-infected mouse serum,metacestode tissue,E.multilocularis-derived EVs,germi-nal layer cells,and protoscoleces.Moreover emu-miR-10-5p was confirmed to be present in higher amounts in E.multilocu-laris-derived EVs,according to RT-qPCR.Further studies confirmed that overexpression of emu-miR-10-5p promoted HSC proliferation and increased α-SMA,collagen Ⅰ,and collagen Ⅲ mRNA and protein expression levels in HSCs.In contrast,EVs extracted from culture supernatants after emu-miR-10-5p knockdown in vesicles did not increase a-SMA,collagen Ⅰ,and colla-gen Ⅲ mRNA and protein expression levels.E.multilocularis vesicle-derived EVs and emu-miR-10-5p thus promote the acti-vation of HSCs.

To evaluate the immunogenicity of inactivated SARS coronavirus (SARS-CoV), three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant, adjuvant,and normal saline respectively. Eight batchs of serum were sampled from the auricular vein at day 7 to day 51, and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test. Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining. The results showed that, rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity, which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses. The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively. In the experimental group, no obvious histopathological changes was detected in the H&E stained slides of heart, spleen, kidney and testis samples, but the livers had slight histopathological changes, and the lungs presented remarkable histopathological changes. These findings are of importance for SARS-CoV inactivated vaccine development.

OBJECTIVE: To study DNA quantification and STR typing of samples pre-treated with pyramidon. METHODS: The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. RESULTS: In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. CONCLUSION Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology* ; Blood Stains ; DNA/isolation & purification* ; DNA Fingerprinting ; Forensic Medicine ; Humans ; Polymerase Chain Reaction ; Reproducibility of Results ; Specimen Handling

OBJECTIVETo study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).

METHODSWe established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.

RESULTSThe prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.

CONCLUSIONThe ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.

Animals ; Apomorphine ; Carcinogens ; Diethylstilbestrol ; Erectile Dysfunction ; etiology ; Humans ; Male ; Myocytes, Smooth Muscle ; ultrastructure ; Nitric Oxide Synthase Type I ; metabolism ; Organ Size ; Penile Erection ; Penis ; enzymology ; ultrastructure ; Pituitary Neoplasms ; chemically induced ; complications ; Prolactin ; blood ; Prolactinoma ; chemically induced ; complications ; Rats ; Rats, Wistar ; Testosterone ; blood

AIM:To evaluate the efficacy and safety of "one-stitch anastomosis through the skin" repair of canalicular laceration.METHODS:The data of 32 cases (32 eyes) of canalicular laceration who underwent repair of lacerated canaliculi with one-stitch anastomosis through the skin were retrospectively reviewed, inferior canalicular laceration in 29 patients,superior canalicular laceration in 1 patient, 2 cases involving both the inferior and superior canalicular laceration. All the operations were performed under surgical microscope, 5-0 silk sutures were used and silicone tube of 0.8mm diameter was employed in intubation. The stents were left in place for 3 months postoperatively and then removed. The follow-up period was 1 to 36 months.RESULTS: In 32 patients, 28 (88%)patients were cured entirely, 3 (9%)patients were meliorated, and 1 (3%)patient had no effects. A total of 29 patients complied with scheduled follow up 1-36 months (average 12 months) after stent removal, and 3 patients were lost in follow-up. All the patients had got good recovery of eyelid laceration with no traumatic deformity in eyelid and canthus.CONCLUSION: In "one-stitch anastomosis through the skin"repair of canalicular laceration, the cut ends could be anastomosed directly,for there was no suture remained in the wound permanently, so there was no suture-related granuloma which might cause obstruction or stenosis of canaliculi. It was simple, economical ,effective and safe.

OBJECTIVETo investigate the effect of DNA methylation in combination with histone deacetylase inhibitor on transcription regulation of Ras associated domain family gene 1(RASSF1A) tumor suppressor gene and the molecular biological behaviors in U266 cells.

METHODSThe U266 cells were treated with different doses of 5-Aza-2'-deoxycytidine (5-Aza-CdR) and Valproate (VPA) each alone or in combination. Methylation-specific PCR (MSP) was used to detect CpG island methylation in RASSF1A promoter. Quantitative real-time reverse transcription polymerase chain reaction (RQ-PCR) was used to examine the expression of RASSF1A gene in U266 cells. MTT was used for cell proliferation. Cell apoptosis and cell cycle were analyzed by flow cytometry.

RESULTSThe methylation of RASSF1A gene promoter was detected in U266 cells, while there was little RASSF1A gene expressing in the control group. The demethylation effect could be detected in the 5-Aza-CdR treated and combined treatment groups but no in the VPA group. The expression level of RASSF1A was induced by 5-Aza-CdR in a concentration-dependent manner while VPA had no such effect. The expression level of RASSF1A mRNA was increased significantly in the combined treatment group. Higher growth inhibition and apoptosis effects were found in 5-Aza-CdR and VPA combination group than that in 5-Aza-CdR or VPA alone group (P < 0.05). After treatment with 5-Aza-CdR or VPA alone for 72 h, more cells were arrested in G(0)/G(1) phase as conpared with control group (P < 0.05), and even more cells were so arrested in combined treatment group (P < 0.05).

CONCLUSIONDNA methylation and histone deacetylase inhibitor can synergistically induce demethylation of the RASSF1A gene, re-express RASSF1A gene silenced in U266 cells, inhibit the proliferation of U266 cells and induce cell apoptosis.

Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Gene Expression ; Histone Deacetylase Inhibitors ; Humans ; Promoter Regions, Genetic

OBJECTIVETo study the expression of connective tissue growth factor (CTGF) and its significance in sporadic ascending thoracic aortic aneurysm (AAA), and initially to investigate the mechanisms of pathological remodeling in AAA.

METHODSAAA specimens were taken from 18 patients during elective surgical intervention, and 18 control specimens of ascending aorta were obtained from patients undergoing coronary artery bypass surgery. Specimens were stained with HE and Masson to evaluate the arrangement and aggregation of cells and collagen types I and III; immunohistochemistry staining was performed using antibodies directed against markers of CTGF; real-time PCR analysis was performed to quantify the expression level of CTGF and collagen types I and III.

RESULTSPathological results show degradation of elastin and hyperplasia of collagen fibers as well as disordered arrangement of smooth muscle cells in AAA. When compared with controls, protein levels of CTGF were significantly increased [(44 ± 4)% vs. (33 ± 5)%, P < 0.01]. Similar patterns were shown in mRNA levels of CTGF (P < 0.01). Using real-time PCR method, elevated levels (relative expression ratio of mRNA: 10.54/3.8 and 1.79/1.19, respectively; P < 0.01, both) of collagen types I and III were observed. CTGF expression had a correlation with both collagen fibers and aortic aneurysm diameter (r = 0.784, P < 0.01; r = 0.793, P < 0.01).

CONCLUSIONSThese results indicate increased expression of aortic collagen types I and III as well as CTGF in AAA specimens, which is likely to be responsible for the aortic wall pathological remodeling. The expression of CTGF was positively correlated with the aortic diameter. As a cytokines factor can stimulate collagen synthesis, CTGF may be involved in the pathogenesis and progression of AAA.

Aged ; Aorta ; metabolism ; pathology ; Aortic Aneurysm, Thoracic ; metabolism ; pathology ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Connective Tissue Growth Factor ; metabolism ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the effect of enterostomy in treatment of locally advanced rectal carcinoma patients with combined chemoradiotherapy and operation.

METHODSClinical data from 51 cases of locally advanced rectal cancer patients treated with preoperative chemoradiotherapy and operation were analyzed.

RESULTSThirty-three patients (64.9%) got staging down of their cancer after preoperative chemoradiotherapy, and 21.6% of patients (11 cases) had complete pathologic response. Thirty-seven patients received enterostomy, including extraperitoneal sigmoidostomy (29 cases), defunctioning ileostomy (8 cases) and double colostomy (3 cases with colon obstruction during preoperative therapy). One case experienced parastomal hernia and one stomal stenosis and 2 cases parastomal infection after enterostomy. No death of enterostomy occurred.

CONCLUSIONColostomy can reduce the pressure of obstructed intestinal tract and contribute much to the preoperative chemoradiotherapy, ileostomy can protect the distal stoma from leakage in sphincter saving operation. Enterostomy could be selected when needed in the favor of locally advanced rectal cancer patients.

Adult ; Aged ; Chemotherapy, Adjuvant ; Combined Modality Therapy ; Enterostomy ; adverse effects ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Radiotherapy, Adjuvant ; Rectal Neoplasms ; pathology ; surgery ; therapy ; Rectum ; drug effects ; radiation effects ; surgery ; Treatment Outcome