Background Glaucoma is common blinding eye diseases characterized by chronic loss of retinal ganglion cells(RGCs).Currently glaucoma pathogenesis is not completely understood,heat shock protein 27 ( HSP27 )may be associated with the pathogenesis of glaucomatous optic neuropathy. Objective Through the establishment of a rat model of high intraocular pressure,detection of the expression of HSP27 antibody in serum and RGCs apoptosis to investigate the role of HSP27 in RGCs apoptosis. Methods Fifty-one clean Wistar rats were divided into high intraocular pressure group (34 rats)and sham operation group( 17 rats)using a random number table.An animal model of high intraocular pressure was established in the right eye by the application of bipolar underwater electrocoagulation on vein of sclera surface in the experimental group,and rats with conjunctiva incision only without electric coagulation were served as sham operation (control).The intraocular pressure of rats of the both groups including experimental and control rats was measured 1,2,4,6,8 weeks after operation and then the rats were sacrificed.1 ml serum was collected from these rats to determine the concentration of HSP27 antibody.The retinas of the rats were isolated and homogenated for the extraction and analysis of the retinal protein by Western blot.Apoptosis of RGCs were assayed by TUNEL.The use of the experimental animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by Kunming Medical Collegc. Results Intraocular pressure was elevated significantly after modeling and remained a high value during the expcrimental duration,showing a significant difference among the different groups ( F =318.502,P＜0.01 ).However,no significant change in intraocular pressure before and after operation in the sham-operation group ( F =2.076,P ＞ 0.05 ).The serum concentration of HSP27 antibody was increased significantly overtime from 1,2,4,6 to 8 weeks after the surgery applied,with a significant difference among various time points( F =154.221,P＜0.01 ),but no obvious difference was seen in the shamoperation group( F =0.422,P＞0.05 ).Importantly,Apoptosis rate of RGCs in the experimental eyes was significant higher as the time prolong after intraocular elevated(x2=856.12,P＜0.05 ).The positive rate of RGCs apoptosis in the rats with high intraocular pressure group was elevated significantly compared to the rats in the sham operation group(P＜0.05). Conclusions High intraocular pressure induces the expressions of HSP27 in retina and the accumulation of HSP27 antibody in rats serum;the elevated HSP27 is associated with the increased cell death in RGCs.

Objective To identify a detailed biological characterization of mesenchymal stem cells (MSCs) isolated from hu‐man umbilical cord(UC) tissue regarding their morphology ,immunophenotype ,purity and proliferative capacity and establish a rea‐sonably cultured and amplified system .Methods After stripping off arteries and veins ,the remaining parts of umbilical cord were cut into 1 mm3 small sections and cultured with DMEM/F12 containing 10% fetal bovine serum .Adhere cells were obtained and the morphology of the cells was observed under inverted phase contrast microscope .The growth curves of them were drawn by CCK‐8 and the cell cycle and surface antigens (CD29 ,CD73 ,CD90 ,CD105 ,CD31 ,CD14 ,CD34 ,CD45 ,CD11b ,HLA‐DR) were detected by flow cytometry .Results Seven to ten days after primary culture ,adhere cells came out of fragments .The MSCs harvested were a high purity and mainly presented as a fibroblast‐like morphology .UC‐MSCs had a strong ability of proliferation through the cell growth curve .The special surface antigens CD29 ,CD73 ,CD90 ,CD105 were positive expression ,while CD31 ,CD14 ,CD34 ,CD45 , CD11b ,HLA‐DR were negative .More than 80% cells of MSCs were found at G0/G1 phase .Conclusion Human UC‐MSCs could be cultured and proliferated in vitro .

0.05).The cerebral infarcted area in SHR were significantly greater than that in SD rats [(42.6?5.6)% vs(29.5?6.7)%,P

Objective To investigate the clinical significance of CD55,CD59 and Aeromonas hydrophila toxin variant (FLAER) in the diagnosis of anemia and paroxysmal nocturnal hemoglobinuria (PNH).Methods Collected 30 healthy controls,22 cases of PNH,33 cases of aplastic anemia (AA),37 cases of iron deficiency anemia (IDA),45 cases of megaloblastic anemia (MA),30 cases of hemolytic anemia (HA) and 31 cases of myelodysplastic syndrome (MDS) from January 2009 to March 2017,CD55,CD59 and FLAER negative cell ratio of peripheral blood neutrophil of them were detected by multipa rameter flow cytometry.Results The detection rates of FLAER in PNH,AA and MDS groups were higher than those of CD55 and CD59,but there was significant difference in AA (x2 =7.759,5.518,P=0.005,0.019＜0.05).The average CD55,CD59 and FLAER deletion rate in PNH and AA group were significantly higher than those in normal control group and other groups (t=2.163～17.890,P=0.000～0.038＜0.05).The number of FLAER in PNH group was higher than CD59 and CD59 was higher than CD55 with the statistically significant difference (t=2.503 ～ 6.308,P=0.000 ～0.016＜ 0.05).Conclusion CD55,CD59 and FLAER have important value in the diagnosis of PNH and differential diagnosis with other anemia diseases,and can also be used to detect the presence of MDS and AA in patients with PNH.FLAER outperforms CD59,CD59 outperforms CD55.

Background As one of the most common microvascular complication of diabetes in eyes,diabetic retinopathy (DR) is one of the most important cause of blindness.Nuclear factor-kappa B (NF-κB) is involved in the occurrence and development of the disease through the activation of a series of inflammatory cytokines.Objective The present study was to investigate the effects of peroxisome proliferator-activated receptor-gamma (PPAR-γ) excitomotor,rosiglitazone,on NF-κB expression and apoptosis of the retinal ganglion cells (RGCs) in the retina with diabetes mellitus. Methods Ninety SPF male Wistar rats were randomized into normal control group,diabetic control group and rosiglitazone group.Diabetes mellitus was induced by intraperitoneal injection of 50 mg/kg streptozotocin(STZ).Then 3 mg/kg rosiglitazone was intragastricly administered once per day in the rosiglitazonegroup,and the same volume of saline solution was used at the same way in the normal control group and diabetic control group from 3 days after modeling.The rats were sacrificed and the eye cups specimen was made at 4,8 and 12 weeks after usage of drugs.Retinal histopathological examination was performed by hematine-eosin staining,and expression of NF-κB p65 protein in retina and apoptotic index(AI) of RGCs were detected by immunohistochemistry and TUNEL assay,respectively in different time points mentioned above.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State and Technology Commission.Results The blood glucose level was significantly elevated at various time points in the diabetic control group and rosiglitazone group compared with normal control group (P＜0.01 ),and that of the rosiglitazone group was significantly declined in comparison to the diabetic control group (q =0.81,0.82,1.23,P＞ 0.05 ).Normal retinal structure was seen in the normal control group,and edema retinal cell and disorder of retinal layers were exhibited in the diabetic control group.Retinal structure was almost normal in the rosiglitazone group.The NF-κB p65 was expressed weakly in the retina of normal control group,but the expression of NF-κB p65 was significantly elevated in the diabetic control group and rosiglitazone group compared with the normal control group(P＜0.01 ).However,the expression of NF-κB p65(A value)was significantly decreased in the rosiglitazone group compared with diabetic control group at 8 weeks and 12 weeks( q=17.77,15.30,P＜0.01 ).There were a few apoptotic cells in rat retina of the normal control group.Compared with the normal control group,the AI of the diabetic control group and rosiglitazone group was significantly reduced(P＜0.01 ).However,the AI of RGCs in the rosiglitazone group was significantly lower than that of diabetic control group in various time points (q =19.28,27.39,49.92,P＜0.01 ). Conclusions As one of the PPAR-γexcitomotors,rosiglitazone can inhibit apoptosis of RGCs through downregulating the expression of NF-κB in rat retina with diabetes mellitus,indicating a protective effect of rosiglitazone on retina in diabetic rat.

BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.

OBJECTIVETo evaluate the clinical effects of transpedicular eggshell technique in treating thoracolumbar deformity.

METHODSFrom December 2008 to December 2011,36 patients with thoracolumbar deformity were treated with transpedicular eggshell technique. There were 20 males and 16 females with an average age of 45 years old (ranged from 20 to 58). Among them, 5 cases were congenital hemivertebrae deformity, 12 cases were secondary to tuberculotic deformity, 14 cases were post-traumatic deformity with pain, 5 cases were ankylosing spondylitis. Low back pain, living ability, scoliotic Cobb angle were analyzed according to VAS scoring, Oswestry Disability Index (ODI), radiological examination.

RESULTSAverage operative time was 245 min and average bleeding was 1 900 ml in 36 patients. All patients were followed up more than 1 year and obtained bone fusion at 1 year after operation. Preoperative,postoperative at 1 week and 1 year, VAS scoring was 7.2 +/- 1.4, 2.5 +/- 1.0, 1.8 +/- 0.5, respectively; ODI was (72.50 +/- 10.80)%, (42.50 +/- 11.10)%, (22.50 +/- 7.90)%, respectively; kyphosis Cobb angle was (76.31 +/- 2.52) degrees, (23.66 +/- 1.16) degrees, (23.67 +/- 1.16) degrees, respectively; lumbar scoliosis Cobb angle was (71.86 +/- 4.02) degrees, (30.81 +/- 2.33) degrees, (30.82 +/- 2.32) degrees, respectively. Postoperative at 1 week and 1 year,above data had obviously improved than that of preoperative (P < 0.05); and there was no significant difference in Cobb angle between postoperative at 1 week and postoperative at 1 year (P > 0.05).

CONCLUSIONTreatment of thoracolumbar deformity with transpedicular eggshell technique could obtain effective correcting and clinical results.

Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Orthopedic Procedures ; methods ; Thoracic Vertebrae ; abnormalities ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Young Adult

Ultrasound therapy is a new type of therapy technology, there are hyperthermia and high intensity focused ultrasound surgery (HIFUS). Compared with a single frequency system, mutiple frequency system has an additional function to combine power patterns of different frequencies. This function increases the availability of power patterns to treat tumors of various shapes and depths. Therefore, we have in this article proposed a system with the ability to drive ultrasonic phased arrays of multiple resonant frequencies for ultrasound hyperthermia and HIFUS. The results show that this system is able to (1) drive multi-element applicators or phased arrays of a single resonant frequency through the multichannel power amplifiers, (2) concurrently drive transducer with different resonant frequencies, (3) adjust the relative phase and output power of each channel for the scanning ultrasonic focus, and (4) operate each channel with good output stability.
Equipment Design ; Humans ; Neoplasms ; therapy ; Transducers ; Ultrasonic Therapy ; instrumentation

To develop a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Norwalk GII. 4 primers which recognized 6 distinct regions on the RNA-dependent RNA polymerase gene of Norwalk GII were designed and used for LAMP assay. Norwalk GII RNA was amplified under isothermal conditions (65 degrees C) for 120 min, and LAMP results were then judged with naked eye, SYBR Green I staining, electrophoretic analysis and restriction digestion. To evaluate the specificity of the RT-LAMP, 48 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses were tested. To compare the sensitivity of the RT-LAMP with that of conventional RT-PCR, Norwalk GII RNA was serially diluted and amplified by RT-LAMP and RT-PCR, respectively. With 46 fecal specimens of Norwalk GII, observation with naked eyes, SYBR Green I staining and electrophoretic analysis were able to detect the PCR products in the RT-LAMP assay. The specificity of RT-LAMP products was also confirmed by digestion of the RT-LAMP products with restriction enzymes. No RNA amplification was observed in 2 fecal specimens of Norwalk GII and 12 fecal specimens of group A rotaviruses. The specificity of the RT-LAMP assay with regard to RT-PCR were 100% for Norwalk GII. The detection limits of RT-LAMP was 15.6 pg/tube for Norwalk GII and similar to that of a RT-PCR assay. Compared to RT-PCR, the RT-LAMP assay has been proven to be a rapid, sensitive, specific and accurate method for detection of the Norwalk GII in fecal specimens, and that RT-LAMP assay is potentially useful for the rapid detection of Norwalk GII from fecal specimens in outbreaks of infectious diarrhea.
Caliciviridae Infections ; virology ; Feces ; virology ; Humans ; Norwalk virus ; genetics ; isolation & purification ; RNA Replicase ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viral Proteins ; genetics

Carcinoma, Hepatocellular ; metabolism ; Glypicans ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; metabolism ; RNA, Messenger ; biosynthesis ; genetics