BACKGROUND: The commonly used culture methods for primary intervertebral disc cells are type Ⅱcol agenase alone digestion method, and type Ⅱ col agenase combined trypin digestion method. However, the acquired cells are few. OBJECTIVE: To acquire nucleus pulposus cells from human degenerative intervertebral disc using a systemic and simplified method. METHODS: Nucleus pulposus cells were isolated and cultured. The morphology of nucleus pulposus cells was observed under inverted phase contrast microscope every day. Primary and subcultured cellsuspension was applied for the determination of cel viability using trypan blue staining. The cel growth was detected with MTT assay. The cel morphology was observed under laser confocal microscopy. RESULTS AND CONCLUSION: Nucleus pulposus cells were successful y isolated from from human degenerative intervertebral disc using the improved method, and cells were subcultured to passage 3. Primary cells were fusiform shaped, while cells at passage 1 and 2 were triangle or polygonal, which were similar to fibroblasts. When cells almost reached the confluence, the cells showed slabstone-like appearance. Trypan blue staining showed that, the primary cel viability was 99%, and passage 3 cells had 93%-95% viability. The proliferation of nucleus pulposus cells gradual y decreased as the generation increased. Compared with passage 1 cells, passage 2 and 3 cells at logarithmic phase trended to be smoother. The cel morphology observed under laser confocal microscopy was similar to the results under phase contrast microscope. The improved simple method can successful y acquire a variety of cells from human degenerative intervertebral disc, and these cells show fine biological properties.

Child ; Humans ; Muscular Atrophy, Spinal ; blood ; diagnosis ; Polymerase Chain Reaction ; methods

Objective To explore the clinical value of B ultrasound-guided psychro-circulation percutaneous microwave coagulation therapy(PMCT) in the treatment of liver cancer.Methods One hundred and sixteen liver cancer patients with 154 lesions were selected to receive psychro-circulation PMCT in our department from July 2004 to December 2007.The pre-and postoperative alpha fetoprotein(AFP) levels were detected.The diameters and blood flow state of the tumors were measured and compared by use of B ultrasound and computerized tomographic scanning(CT).The postoperative 1,2 and 3 y survival rate of those patients was followed up and recorded.Results No mortality or severe complications occurred in this series.Preoperative serum AFP level of the patients was(421.6?36.5)?g/L and decreased obviously to(232.5?25.6)?g/L at 1 month after psychro-circulation PMCT(P

AIM: To assess the efficacy and safety of prophylactic laser photocoagulation for retinal breaks before laser in situ keratomileusis (LASIK) in myopic eyes.METHODS: From April 2000 to April 2004, totally 1 845 eyes ( 1 233 patients ) requesting LASIK had a fundus examination with indirect ophthalmoscopy before the surgery. They were divided into two groups according to the presence (Group 1) or absence of retinal breaks (Group 2). All patients with retinal breaks, though they were asymptomatic, underwent prophylactic laser photocoagulation to seal the breaks before LASIK.RESULTS: Patient age ranged from 18 to 43 ( 25.3±5.7) yaers old. Mean preoperative spherical equivalent refraction (PSER) was -7.44± 2.13 D (range, -1.50 to -14.50 D). Retinal breaks were identified and treated in 37eyes (2.05%) of 32 patients;1 808 eyes of 1 201 patients had no retinal breaks. No statistical difference was found in age ( P ＞0.05) or gender (P ＞0.05) between the two groups. Significant difference of PSER was noted between Group 1 (-9.41± 4.15D) and Group 2 (-7.52±3.71D) (P＜0.05). During a mean 14mo follow-up, none of the patients developed retinal detachment.CONCLUSION: The efficacy and safety of prophylactic laser photocoagulation for retinal breaks was confirmed.Retinal breaks should be identified and treated by photocoagulation in eyes before LASIK for myopia.

Bronchi ; Bronchoscopy ; Child ; Foreign Bodies ; surgery ; Humans ; Magnets ; Male

Background Oxidative damage may cause the functional dysfunction and death of retinal vascular endothelial cells(RVECs),and further leads to the development of retinal vascular diseases.Fufang xueshuantong has a therapeutic effect on retinal vascular diseases,but little is known about its molecular mechanism.Objective The goal of this study was to investigate the protective effects and mechanism of fufang xueshuantong on injury of human RVECs induced by tert-butyl hydroperoxide(t-BHP).Methods Human RVECs were isolated from healthy donor eyes and primarily cultured and then identified by flow cytometry.The third to fifth generations of cells were used in this experiments.The fufang xueshuantong solution of 0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L were added in the cuhure plate with 5 × 104/L cells respectively in the experimental groups,and t-BHP of 75,100,200 and 300 μ.mol/L were added in the model control groups.MTT was used to detect the A490and survival rate of RVECs.The apoptotic rate and death rate of the cells were evaluated by double staining of Annexin V-FITC/PI.Morphology of human RVECs were examined using invert microscopy and Hoechst33258 staining.The expressions of nitro tyrosine (a marker of oxidative damage of protein)and 8-OHdG(a marker of oxidative damage of DNA)in human RVECs were assessed by the immunofluorescence staining.Western blot was used to detect the expressions of nuclear factorkappa B(NF-KB),p53,bcl-2 and bax after 6,12,24 hours t-BHP action.This study was approved by the Ethic Committee of Zhongshan Ophthalmic Center.Results No significant difference was found in A490value among the normal control group,0.0625,0.1250,0.2500,0.5000 and 1.0000 g/L fufang xueshuantong groups(F =1.989,P＞0.05).The survival rates of the cells were lower in 75,100,200 and 300 μmol/L t-BHP groups compared with corresponding fufang xueshuantong groups(t =14.57,13.82,21.51,32.64,P＜ 0.01).The percentages of normal cells were evidently lower in 75,100,200 and 300 μmol/L t-BHP compared with corresponding fufang xueshuantong groups(t=14.908,5.495,17.165,26.330,P＜0.01).The numbers of deformation and death of the human RVECs increased as the elevated concentration of t-BHP,but those in fufang xueshuantong groups were less than the t-BHP groups under the invert microscopy.Compared with t-BHP groups,the expressions of nitro tyrosine,8-OHdG,NF-KB,p53 and bax were lower but the expression of bcl-2 was higher in human RVECs with the statistically significant differences(P＜0.05).Fufang xueshuantong at the concentration of 0.2500 g/L showed maximally protective effect on human RVECs.Conclusions Fufang xueshuantong protects human RVECs against the t-BHP-induced injury through downregulating the expression of NF-kB,p53,bax and up-regulating the express of the bcl-2 protein.

Objective To investigate the efficacy and safety of concurrent chemoradiation with S -1 and cisplatin for the treatment of locally advanced mid and upper thoracic esophageal carcinoma.Methods Forty -six patients with locally advanced mid and upper thoracic esophageal carcinoma were enrolled in this study and received two cycles of S-1 60 mg · m-2 · d -1 for 14 days and then rested for 7 days;cisplatin 25 mg · m-2 were divided into three times.Meanwhile, concurrent radiotherapy (60 Gy total, 2 Gy a day) was done.When the radiotherapy finished, patients received two more cycles of chemothera-py.The data of clinical efficacy and side effects were analyzed.Results Therewere 26 patients ( 56.5%) diagnosed with T4 and 30 patients (65.2%) with N1.The clinical response rate was 67.4%, the estimated overall survival and progression -free survival rates after 2 years were 54.5% and 45.2%, respectively.The primary adverse reactions (≥grade 2) included radiation-related esophagitis (28.3%) and neutropenia ( 37.0%).No treatment -related death occurred.Conclusion Concurrent chemoradiation with S -1 and cisplatin for the treatment of locally advanced mid and upper thoracic esophageal carcinoma exhibited encouraging results with clinically manageable adverse reactions.

Objective To explore the distinctive ultrasonographic characters of enthesitis in patients with spondyloarthritis (SpA) and evaluate the diagnostic value of ultrasonography in SpA by comparing ultrasonography of enthesis in the lower limbs of patients with SpA,rheumatoid arthritis (RA) and osteoarthritis(OA).Methods Entheses in the lower limbs (quadriceps tendon,proximal patellar ligament,distal patellar ligament,achilles tendon and plantar aponeurosis) were detected in SpA,RA,OA patients and health controls by ultrasonographic examination.Results A total of 46 SpA,23 RA and 12 OA patients were enrolled in this study,with 26 SpA and 7 RA patients complaining of heel pain(P ＜0.05).There were 126 enthesal sites(126/460,27.2％) were detected abnormal in SpA group,compared with 54 of 230 (23.5 ％) sites in RA group and 27 of 120 (22.5 ％) sites in OA group (P ＞ 0.05).Sixty five (70.7 ％) sites of the distal patellar ligaments were abnormal in SpA group,while that were 19(41.3％)in RA group(P ＜ 0.05)and 13 (54.2％)in OA group.Twenty six sites of tendon thickness and 22 sites of bone erosion in the distal patellar ligaments were found in SpA group,while only one site of each in RA group(P ＜ 0.05) and 4 sites of tendon thickness in OA group (P ＜ 0.05) were found.Tendon thickness,bursal synovitis and bone erosion coexisting in one distal patellar ligament indicated SpA,the sensitivity,specificity and the area under the curve (AUC) of which were 70.7％,65.5％ and 0.740,respectively.Conclusion Tendon thickness and bone erosion of entheses were more likely the distinctive ultrasonographic characters of enthesitis in SpA patients.Tendon thickness,bursal synovitis and bone erosion in the distal patellar ligament could be an ideal tool for SpA diagnosis.

Contemporary hospital information systems normally contain multiple applications,which are traditionally deployed as one application per server.More servers in the system will incur high system costs,low server efficiency and poor management and maintenance.The information system reform project of Wuxi Maternal and Child Health Hospital called into play the virtualization based on VMware.The project brought forth a number of merits,namely sizably raising efficiency of servers integration,simplifying server group management,reducing cost and promoting hospital informationization.

Objective To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy(X-ALD).Methods The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells.Maternal contamination was evaluated by paternity test.PCRRFLP,sequencing and denaturing high performance liquid chromatography(DHPLC)were used to detect the ABCD1 gene of fetal genome.Results In the pedigree 1,the PCR product(799 bp)of the fetus 1 and her father(normal control)could be digested with BcnI. No P560L mutation,which was present in the index patient,was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing.In the pedigree 2,the PCR product(232 bp)of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation,which was present in the propositus,was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing.In the pedigree 3,the PCR product(271 bp)was digested with AciI.the pattern of the fetus 3 and the propositus being the same,and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing.In the pedigree 4,the PCR product(269 bp)was analyzed with the DHPLC,and the pattern of elution peaks of the fetus 4 and her father was similar,but different from that of the propositus.No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing.Judging from the sex of the fetuses,fetuses 1 and 2 were normal homozygotes,fetus 3 was an ALD hemizygote,and fetus 4 was a normal hemizygote.Conclusion A new protocol for X-ALD prenatal molecular diagnosis is proposed,which would ensure the accuracy of prenatal diagnosis.