? AIM: To investigate the influence of phacoemulsification on corneal endothelial cells and its injury risk factors in diabetic cataract patients.?METHODS: From January 2013 to October 2015, 186 diabetic cataract patients ( 224 eyes ) as diabetes group and 190 patients with simple cataract ( 227 eyes ) as control group in our hospital were enrolled. All patients received phacoemulsification combined with intraocular lens implantation. Observation of corneal endothelial cell density, coefficient of variation and percentage of hexagonal cells preoperatively, 1d, 1wk, 1 and 3mo postoperatively were carried out, and multiple Logistic regression analysis for risk factors of corneal endothelial cell injury was taken.?RESULTS: There were no significant difference in the density of corneal endothelial cells, the coefficient of variation and the percentage of hexagonal cells between the two groups before surgery (P>0. 05). Each time point after operation corneal endothelial cell density and the percentage of hexagonal endothelial cells of diabetes group were significantly lower than the preoperative and those of control group ( P < 0. 05 ). Each time after operation endothelial cell coefficient of variation of diabetes group were significantly higher than the preoperative(P<0. 05). The endothelial cell coefficient of variation in diabetes group of the 1wk, 1 and 3mo after operation were significantly higher than those of the control group ( P<0. 05 ) . Corneal endothelial cell density and percentage of hexagonal cells of the control group at 1wk, 1 and 3mo after operation were significantly lower than the preoperative ( P<0. 05 ). The endothelial cell coefficient of variation of control group at 1wk, 1 and 3mo after operation were significantly higher than the preoperative(P<0. 05). Single factor analysis showed that age, shallow anterior chamber, long ultrasonic time, short eye axis, high energy ultrasonic, high cumulative released energy, a lot of perfusate and nuclear hardness ≥ grade Ⅲ associated with corneal endothelial cell injury ( P< 0. 05 ) . Multivariate Logistic regression analysis showed that age, long ultrasonic time, high ultrasonic energy, high cumulative released energy and nuclear hardness ≥ grade Ⅲ were the risk factors of corneal endothelial cell injury, the OR value and 95%CI were 1. 742 (1. 056-2. 682), 1. 958 (1. 227-3. 135), 2. 064(1. 274-3. 256), 2. 585(1. 493-3. 682), 2. 193 (1. 348-3. 316).?CONCLUSION: The injury of corneal endothelial cells after phacoemulsification in diabetic cataract patients is more serious than in patients with simple cataract. Age, long ultrasonic time, high ultrasonic energy, high cumulative released energy and nuclear hardness ≥grade Ⅲ are the risk factors of corneal endothelial cell injury.

AIM: To study the role of nitric oxide ( NO ) and γ-aminobutyric acid ( GABA) in the formation of amblyopia by establishing 2 different types of amblyopic models.METHODS:A total of 18 aged 3-week kittens were randomly divided into monocular deprivation, strabismus and normal groups. All types of amblyopia were developed in the experimental eyes that were detected by P-VEP 12wk later. The cats were killed and the immunocytochemistry staining method were applied to observe under the light microscope the changes of distribution and positive cells areas of NO and GABA across the amblyopic retinal, compared to that from the normal cats of identical age.
RESULTS: The P-VEP showed that the amplitude of wave P1 was lower (P<0. 05) and the P1 latent time was longer ( P<0. 05 ) in two types of amblyopic cats than those in the normal cats. Compared to the normal cats, the NO and GABA positive cells areas were obviously reduced ( P<0. 05 ) across the retina in the amblyopic cats. But no significant difference was found between two kinds of amblyopic cats.
CONCLUSION:The NO and GABA play an important role in the formation of amblyopia in the level of retinal.

Objective To discuss the significance of serum IgE before and after inhale glucocorticoid treat-ment of children's asthma. Methods 520 children with asthma were seleceted from the outpatient. Different type of fluticasone propionate were given to different age groups: Aerosol type by a spacer in less than 5 years old,and in-halant (Seretide) 5 years and the above. The dosage was between 200 μg/day to 375 μg/day. IgE was tested before and 3 months after the treatment. Results Serum IgE decreased significantly in 3 months treatment [ from (496.12±24.75) kU/L to (390.71±18.71) kU/L] (t=7.337,P<0.01). The change of IgE was related to clinical effect and age. The level increased in those less than 3 years [(307.05±34.71)kU/L vs (483.09±41.78) kU/L] (t=2.963,P=0.004),but decreased between 4 to 5 years old group [(543.46±51.03) kU/L vs (316.93±29.30) kU/L] (t=3.368,P=0.000) ,and decreased between 6 to 14 years old group[ (586.30±37.19)kU/L vs (387.61±27.60) kU/L] (t=4.827,P=0.000). In fluticasone group IgE level changed from (468.91±32.81) kU/L to (359.03±22.79) kU/L after treatment (t=5.988,P<0.01),which decreased from (586.30±37.19) kU/L to (387.6±27.60) kU/L in Salmeterol group (t=4.827,P<0.01). In 260 cases of IgE below 300 kU/L 109 cases (41.92%,109/260) increased while in 260 cases of IgE above 300 kU/L,total IgE lev-el increased in 45 cases (16.15% ,45/260) after treatment,with statistical significance(χ<'2>=37.789,P=0.000). Conclusion Inhale glucocorticoid can make the level of IgE decreased.

Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.

Aim To study the effect of PA on the proliferation and maturation of cord blood derived DC in vitro. Methods The monocytes were isolated from human umbilical cord blood and cultured with PA, GM-CSF+IL-4+TNF-?(GTI), GTI+PA (GTIP), respectively. On day 7 of cultivation, the CD1a, CD83,HLA-DR and CD34 antigens expression of cells were analyzed by immunohistochemistry. Result The cells with typical morphological properties of DC can be observed with electron microscope among PA, GTI and GTIP groups. The CD1a and CD83 positive rates of PA group increased significantly, reaching 19.63%?3.61% and 14.52%?5.79% respectively, much higher than that of the control. In addition, compared with GIT group, the positive rate in GTIP group has increased remarkably. Conclusion PA does not only promote the proliferation and maturation of cord blood derived DC, but also cooperate with GTI to enhance the production of DC. PA is a useful activator of DC.

Objective:To express and purify the TRIM5? chimaera[TRIM5? H(R328-332)] protein and to explore the interaction between the TRIM5? H(R328-332)and HIV-1gag. Methods:The plasmid pET28aTRIM5? H(R328-332) was transformed to E.coli BL21 (DE3) strain ,and the expression of TRIM5? H(R328-332) protein was induced by IPTG,purified with Ni2+ chromatography.The expression and purification of TRIM5? H(R328-332) were analyzed by SDS-PAGE and Western blot,and the interaction between TRIM5? H(R328-332) and HIV-1gag was detected by co-immunoprecipitation,His pull-down and ELISA. Results:The recombinant plasmid pET28aTRIM5? H(R328-332) was successfully expressed in E.coli. The results showed that the purified full length TRIM5? H(R328-332) interacted with HIV-1gag protein. Conclusion:The human TRIM5? chimaera was expressed successfully in vitro,and the study demonstrates that the human TRIM5? chimaera interacts with HIV-1 gag in vitro.

Objective To approach the effect of Shenfu injection (SFI) and conventional early goal-directed therapy (EGDT) on organ functions and outcomes of septic shock patients. Methods Eighty-four cases conformed to the criteria for the diagnosis of septic shock admitted to Department of Critical Care Medicine of Xuzhou Central Hospital were randomly divided into conventional treatment group (42 cases), and SFI treatment group (42 cases). Conventional treatment was given in the two groups;in SFI treatment group, SFI 100 mL was additionally given by trace continuous intravenous pump 20 mL/h, twice daily for 7 days. Before and after treatment for 1, 6, 12, 24, 48, 72 hours, the levels of hemodynamic status, lactic acid and dosage of vasoactive drugs used, organ function, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, sequential organ failure assessment (SOFA) score, the time of weaning from ventilator, the length of stay in intensive care unit (ICU), time without organ failure and 28-day mortality rate were observed. Results Compared with those before treatment, after treatment in the two groups, the mean arterial pressure (MAP), cardiac index (CI) and systemic vascular resistance index (SVRI) were increased, while the levels of heart rate (HR) and lactate were decreased (all P<0.05). Compared with conventional treatment group, in SFI treatment group, after treatment for 24 hours, the MAP level was increased significantly [mmHg (1 mmHg=0.133 kPa):75.40±9.75 vs. 71.80±11.08, P < 0.05], that continued to 48 hours; after treatment for 6 hours, the CI level was increased obviously (mL·s-1·m-2: 75.18±34.84 vs. 67.35±39.34, P < 0.05) , that continued to 48 hours; after treatment for 6 hours, the lactic acid level was decreased markedly (mmol/L: 2.03±0.82 vs. 2.24±0.97, P < 0.05);in the comparison of dosage of vasoactive drugs used between two groups, the difference was not significant (all P >0.05). Compared with that before treatment, in the conventional treatment group after treatment for 1 and 3 days, gamma glutamyl transpeptidase (GGT) was increased, on the 5th day it began to decrease, reaching its minimum on the 7th day (U/L:26.75±16.74 vs. 46.96±25.85);while in SFI treatment group, GGT was increased after treatment for 1 day, on 3rd day it began to decrease, reaching its lowest level on the 7th day (U/L:22.41±17.87 vs. 51.23±27.74);aspartate aminotransferase (AST), total bilirubin (TBil), oxygenation index (PaO2/FiO2) were increased after the treatment for 1, 3, 5, 7 days, and blood urea nitrogen (BUN), creatinine (Cr) were decreased at different time points after treatment. In the conventional treatment group, the precursor protein (PA) was decreased after treatment for 1, 3, 5 days, on the 7th day it was increased (mg/L:134.20±63.44 vs. 115.70±45.96);while in SFI treatment group, after the treatment for 1 days and 3 days, it was decreased, on the 5th day it was increased, reaching its highest level on the 7th day (mg/L:145.40±59.75 vs. 108.20±54.34). Compared with those before treatment, after treatment for 1, 3, 5, 7 days, APACHEⅡscore and SOFA score were decreased in the two groups, but there was no statistically significant difference in APACHEⅡscore between the two groups, in SFI treatment group after treatment for 3 days, SOFA score was significantly lower than that of the conventional treatment group (6.31±3.86 vs. 7.14±4.03, P<0.05), that continued to the 7th day after treatment. In SFI treatment group, the time for weaning from ventilator (days:7.5±3.5 vs. 9.1±3.2) and the length of stay in ICU (days: 16.1±9.2 vs. 18.7±8.3) were significantly shorter than those in conventional treatment group (both P<0.05). There were no significant differences in time without organ failure (days:14.5±4.2 vs. 15.3±3.1) and 28-day mortality rate [28.6%(12/42) vs. 31.0%(13/42)] between SFI group and conventional group (both P>0.05). Conclusion The combined use of SFI and EGDT can improve hemodynamics, reduce damage to vital organs, and shorten the times for ventilation and stay in ICU in septic shock patients.

Objective To observe the expression of serine/threonine kinase 31 (STK31) in osteosarcoma and its effect on the malignant biological behavior of osteosarcoma.Methods Fifteen cases of osteosarcoma specimens and adjacent normal tissue were collected.The expression of STK31 in tumor tissues and normal tissue were detected by immunohistochemistry,real-time quantitative PCR and Western blot.The STK31 knockout plasmids PGenesil-STK31-shRNA or control plasmid pGenesil-1 were transfected into osteosarcoma cell line MG63 cells.The effect of STK31 on the proliferation of MG63 cells was detected by CCK8 cell activity assay.Tanswell experiment was used to observed the effect of STK31 on the migration ability of osteosarcoma cells.Results Immunohistochemical showed that STK31 expressed in the tumor tissue,and it was significantly higher than the adjacent normal tissues;Real time quantitative PCR[(3.65±0.83)vs.(1.05±0.14),P＜0.05] and Western blot also revealed that STK31 expression in tumor tissue were significantly higher than adjacent normal tissues(P＜0.05);CCK8 experiments showed that knockdown STK31 inhibited proliferation of MG63 cell when compared with the control group after 36 h[(1.71±0.17)vs.(1.39±0.11),P＜0.05],72 h[(2.15±0.21)vs.(1.54±0.14),P＜0.05];Tansewell experiments showed that transfection of pGenesil-STK31-shRNA could suppress MG63 cell's migration[(13±4)vs.(55±8),P＜0.05].Conclusion STK31 is overexpression in osteosarcoma with increased biological activity of osteosarcoma cells.

Objective To explore the clinical characteristics and influencing factors of primary nephrotic syndrome (PNS) combined with hypercoagulability in children. Methods The clinical data of 57 children with primary PNS were analyzed retrospectively. The clinical features and treatment were compared among high coagulation state group, non high coagulation state group and control group (20 children). At the same time, the differences between the simple nephrotic syndrome group (SNS) and nephritic syndrome group (NNS) in hypercoagulable state were analyzed. In addition, the correlation analysis was performed. Results Among 57 patients, there were 50 patients in high coagulation state group and 7 in non high coagulation state group. There was no significant difference in gender, age and clinical manifestations between two groups (P>0.05). The platelet (PLT) count, platelet aggregation (PCT), albumin (Alb), fibrinogen (Fib), D-dimer (D2) were significantly higher than those in the control group, and there were statistically significant differences (P all<0.01). There were significant differences in the levels of PLT, Fib, D2 and complement C4 between hypercoagulable state group and non hypercoagulable state group (P all<0.05). There were significant differences in HCT, TC, LDL, PT and complement C3 levels between SNS group (n = 32) and NNS group (n =18) in 50 patients with high coagulation state (P<0.05). There was positive correlation between HCT and complement C3 (r=0.30, P<0.05), while there was no correlation between PLT and other indices (P>0.05). All of the 57 patients were improved and has no thrombosis after the treatment. Conclusion Children with primary PNS were usually associated with different degrees of hypercoagulable state, and PLT, Fib, D2 could be used as reference indices for the severity of hypercoagulable state, and the activation of complement system might be related to the occurrence and development of hypercoagulable state.

OBJECTIVETo observe the effects of electroacupuncture (EA) on circadian rhythm of temperature and melatonin (MT) in depression rats model induced by chronic stress, so as to explore the biological mechanism of EA for depression.

METHODSTwenty-four SD rats were randomly divided into a control group, a model group and an EA group, 8 cases in each one. Rats in the control group were treated with normal diet for 21 days without any treatment. In the model and EA group, rat model was established by chronic unpredictable stress combined with solitarily feeding method, and rats in the EA group was treated with EA at "Baihui" (GV 20), "Yintang" (GV 29) 1 h before stress stimulation everyday, 2 Hz in frequency and intensity was favorable with the head of rat slightly shivering. The needles were retained for 20 min, once a day for totally 21 days. After EA treatment, open-field experiment was adopted to observe the behavioral improvement; the rats temperatures were monitored at six time points (2:00, 6:00, 10:00, 14:00, 18:00, 22:00) and orbital blood sampling was collected. The level of serum MT was tested by enzyme linked immunosorbent assay. The circadian rhythm changes of temperature and serum MT in each group were compared.

RESULTSThe numbers of horizontal movement and vertical movement in the model group were obviously lower than those in the control group (both P < 0.05), while those in the EA group were significantly improved compared with those in the model group (both P < 0.01). The circadian rhythm of temperature and MT disappeared in the model group, which was improved into normal level after EA treatment.

CONCLUSIONThe electroacupuncture has regulation effects on circadian rhythm of temperature and melatonin in depression rat model induced by chronic stress.

Animals ; Circadian Rhythm ; Depression ; metabolism ; physiopathology ; therapy ; Electroacupuncture ; Humans ; Male ; Melatonin ; metabolism ; Rats ; Rats, Sprague-Dawley