Objective To evaluate the effect of immune fluorescence chromatography on glycosylated hemoglobin (HbA1c). Methods The precision of immune fluorescence chromatography was evaluated with samples of 6.0% and 8.0% fixed value. Group of High performance liquid chromatography (HPLC) as control, HbA1c for 200 samples of EDTA-K2 anti-coagulated whole blood were detected by immune fluorescence chromatography to synchronous blinded trial. Results As to the precise of immune fluorescence chromatography in the samples of 6% and 8%, values of coefficient of variation were 5.1% and 5.3%, respectively. The linear regression equation of immune fluorescence chromatography and HPLC was Y=-0.110+1.021X and the correlation coefficient was 0.982. 6.0% and 8.0% as the cut-off value, kappa values were 0.950 (P < 0.001) and 0.922 (P < 0.001), respectively. Conclusion Immune fluorescence chromatography and HPLC is consistent with detection of HbA1c, which can be used for clinical detection of HbA1c.

OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.

METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.

RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.

CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

Objective To observe the variation of phosphatidylinositol-3 kinase (PI3K),protein kinase B (AKT),sodium iodide symporter (NIS) mRNA and protein expression in rat mammary tissues and serum insulin growth factor Ⅰ (IGF-1) under different iodine nutrition levels,and to study the role of PI3K-AKT signaling pathway in the process of mammary gland intaking iodine during lactation period.Methods Totally 130 Wistar rats (100 female rats,30 male rats) were randomly divided into five groups with 20 female rats in each group:①control group (NI):was feed with normal diet and iodine content 50 μg/L in deionized water;②low iodine group 1 (LI1 group):was feed with low iodine diet and deionized water;③low iodine group 2 (LI2):was feed with low iodine diet and iodine content 5 μg/L in deionized water;④high iodine group 1 (HI1 group):was feed with normal diet and iodine content 3 000 μg/L in deionized water;⑤high iodine group 2 (HI2):was feed with normal diet and iodine content 10 000 μg/L in deionized water.After feeding for 3 months,females were mated with male rats,then male rats were taken out and every female rat was feed individually.Urinary iodine level of rats in lactation period 10 days after giving birth was tested.Blood and mammary tissue samples of rats in lactation period were taken after killing them.Enzyme linked immunosorbent assay (ELISA) was used to detect serum IGF-1 level,real-time fluorescence quantification PCR to detect the mRNA expression of mammary gland PI3K,AKT and NIS,Western blotting to detect mammary gland PI3K,total AKT,phosphorylation AKT (p-AKT) and NIS protein expression.Results The medians urinary iodine of lactation period rats in LI1 and LI2 (3.16,6.36 μg/L) were significantly lower than that in NI group (162.59 μg/L),and were significantly higher in HI1 and HI2 (2 356.27,11 507.29 μg/L) than that in NI group.The differences were statistically significant (all P ＜ 0.01).Compared with control group [(8.84 ± 2.12) μg/L],the content of serum IGF-1 increased significantly in lactation period rats in LI1 and LI2 groups [(13.30 ± 2.37) and (10.90 ± 1.92) μg/L,all P＜ 0.01].The real-time fluorescence quantification PCR detection results indicated that the differences were statistically significant by comparing NIS,AKT,PI3K mRNA expression of the mammary tissues of lactation period rats in the five groups (F=14.916,36.477,14.994,all P＜ 0.01).Among them,NIS mRNA expression quantities in LI1 and LI2 groups (0.75 ± 0.40,0.89 ± 0.51) were significantly higher than that in NI group (0.53 ± 0.31),and significantly lower in HI2 group (0.30 ± 0.24) than that in NI group (P ＜ 0.05 or ＜ 0.01).AKT mRNA expression quantities in LI1 and LI2 groups (0.90 ± 0.19,0.64 ± 0.22) were significantly higher than that in NI group (0.43 ± 0.22),and significantly lower in HI2 group (0.29 ± 0.15) than that in NI group (P ＜ 0.05 or ＜ 0.01).PI3K mRNA expression quantity in LI1 group (0.85 ± 0.42) was significantly higher than that in NI group (0.50 ± 0.24),and significantly lower in HI2 group (0.28 ± 0.10) than that in NI group (all P ＜ 0.01).Western blot detection results indicated that the differences were statistically significant by comparing mammary gland NIS protein expression of lactation period rats in the five groups (F=4.510,P＜ 0.01).Among them,LI1 group (1.67 ± 0.97) was significantly higher than NI group (0.87 ± 0.43,P ＜ 0.05).The differences were statistically significant by comparing the p-AKT protein expression among groups (F =3.528,P ＜ 0.05).Among them,HI2 group (1.10 ± 0.30) was significantly higher than NI group (0.75 ± 0.23,P ＜0.05).The differences were not statistically significant by comparing total AKT and PI3K protein expression among groups (F =0.558,1.319,all P ＞ 0.05).Conclusion The inhibitory effect of PI3K-AKT signaling pathways on NIS in the mammary gland was weaker than the effect of iodine intake.But the expression of functional p-AKT was gradually increased with the increment of iodine intake,which had been presented inhibit effect on NIS expression in lactating mammary gland.

AlM:To compare the efficacy and safety of latanoprost and brimonidine in the treatment of open angle glaucoma, and provide reference for rational drug use.METHODS:A total of 121 cases ( 136 eyes ) who were diagnosed as primary open angle glaucoma were selected in this study, and they were randomly divided into experimental group (62 cases, 70 eyes) and control group ( 59 cases, 66 eyes) according to different drug treatment. Patients in the control group received brimonidine eye drops twice a day, while patients in the experimental group received latanoprost eye drops once a day. The intraocular pressure, visual acuity and adverse reactions were checked of the two groups in the following 3mo.RESULTS:The intraocular pressure of patients in the control group was 18. 1 ± 1. 3mmHg, while the experimental group was 17. 0 ± 0. 9mmHg after 12wk of treatment, which were both lower than before (P<0. 05). The fluctuation of intraocular pressure in the experimental group was significantly lower than that of the control group. There was no significant difference in the LogMAR visual acuity between before and after treatment in the control group, while the LogMAR visual acuity of the experimental group was significantly improved. The control group had hyperemia, burning sensation, tearing, eyelid edema and other adverse side effects, and the experimental group had little adverse reactions. CONCLUSlON: Latanoprost can significantly reduce intraocular pressure in glaucoma patients with in the follow- up time, and reduce the impact of elevated intraocular pressure in the vision of glaucoma patients, with little adverse reaction, worthy of clinical application.

AIM: To investigate the efficiency of intravitreal ranibizumab therapy ( IVR ) for polypoidal choroidal vasculopathy ( PCV) in single or multiple polyps.
METHODS: A total 63 patients diagnosed with PCV in Shaoxing City People's Hospital from May 2013 to May 2015 were enrolled and divided into single polyp group and multiple polyps group. All patients received intravitreal ranibizumab 3 monthly and were followed up for 12mo. Observe the changes of best corrected visual acuity ( BCVA ) and central retinal thickness ( CRT ) at different time points.
RESULTS: The single polyps group exhibited a better BCVA, shorter greatest linear dimension, and lower prevalence of fibro - vascular pigment epithelial detachment compared with the multiple polyp group before treatment (P<0. 05). Significant difference of BCVA were observed at 3, 6 and 12mo between the two groups (P<0. 05). BCVA at 3, 6, 12mo was significantly better than that at baseline in single polyps group. The single polyp group exhibited a significantly thinner CRT at 6 and 12mo compared with multiple polyps group (P<0. 05). The single polyp group showed improvement in CRT over the followed up period(P<0. 05). The CRT in multiple polyps group at 3 and 6mo significantly decreased compared with preoperative (P<0. 05).
CONCLUSION: IVR meet better result in PCV patients with multiple polyp and polyp numbers may be valuable to prognosis.

Objective:To investigate the relationship between FA,ET-1,Hcy and VEGF in patients with pregnancy induced hypertension.Methods:Selected 80 cases of patients with pregnancy induced hypertension who were treated in our hospital from January 2013 to September 2016,divided into A group (40 cases),and preeclampsia and preeclampsia B group (40 cases),60 cases of normal late pregnant women as C group,60 cases of normal non pregnant women as D group.The plasma levels of FA,ET-1,Hcy and VEGF in each group were detected,and the correlation analysis was performed.Results:The plasma levels of FA and VEGF of group B were lowest in the four groups (P＜0.05),the plasma ET-1 and Hcy levels of group B were highest (P＜0.05).The plasma FA and VEGF levels of women with pregnancy induce hypertension were lower than those of the healthy late pregnant women.The plasma ET-1 and Hcy levels were higher (P＜0.05).The multivariate logistic regression analysis showed that FA,ET-1,Hcy and VEGF levels were risk factors for pregnancy induce hypertension.Conclusion:The elevation of plasma Hcy level caused by FA deficiency in pregnancy might be related to the development of the disease and the incidence of pregnancy induced hypertension syndrome,VEGF might be an important factor in the occurrence and development of pregnancy induced hypertension syndrome.

Objective To investigate the application value of MRS in temporal lobe epilepsy (TLE) without hippocampal sclerosis.Methods A retrospective study was conducted in 23 unilateral TLE patients (TLE group) with absence of hippocampal sclerosis and 20 age-matched normal control subjects (control group).All of them underwent conventional MRI and MRS.N-acetyl aspartate (NAA)/creatine (Cr),choline (Cho)/Cr ratios of bilateral hippocampus in 2 groups were analyzed.Eleven of 23 patients underwent surgical treatment,pathological findings and surgical outcomes were evaluated.Results The NAA/Cr ratios of ipsilateral hippocampus significantly decreased compared with that in the contralateral hippocampus (t=-7.97,P＜0.001) and the control group (t=-9.96,P＜0.001).There was no significant difference between the contralateral hippocampus in patients and the control group (t=-1.21,P=0.12).The Cho/Cr ratios of ipsilateral hippocampus in TLE had no significant difference compared with contralateral hippocampus (t=0.50,P=0.31) and the control group (t=-0.59,P=0.28).The pathological findings of the 11 patients who underwent temporal lobe resection indicated small amounts of neuronal loss and unobvious gliosis.Conclusion MRS is helpful in clinical practice to lateralize and localize the epileptogenic foci in the absence of hippocampal sclerosis in patients with TLE.

In gene manipulation, different selectable markers with various linkers are necessary. In order to get selectable markers directly, we constructed from pBlueScript SK(-) a series of particular plasmids, pSKsymKm, pSKsymBle, pSKsymEry, pSKsymHyg and pSKsymGm, each contains Kanamycin, Bleomycin, Erythromycin, Hygromycin or Gentamycin resistance cassette. By restriction enzyme digestion and gel extraction, any of five antibiotic resistance genes with specific ends can be conveniently obtained.

Objective To analyze the correlation between clinical features and renal dysfunction in patients of acute lacunar infarction with progressive cerebral microbleeds (CMBs). Methods Two hundred and sixty-five patients with first-episode acute lacunar infarction were selected. The serum creatinine was measured within 24 h of admission and the estimated glomerular filtration rate (eGFR) was calculated. The brain MRI (including gradient-echo images) was examined within 2 d of admission and after 1 years of follow-up, respectively. The progressive CMBs was assessed with microbleeds anatomical rating scale (MARS), and the patients were divided into progressive CMBs group (progressive group, 42 cases) and non progressive CMBs group (non progressive group, 223 cases). The clinical features of 2 groups were compared and the correlation between progressive CMBs and renal dysfunction was analyzed. Results The age, 24 h pulse pressure, incidences of renal dysfunction and CMBs in progressive group were significantly higher than those in non progressive group: (69.8 ± 5.8) years vs. (61.5 ± 4.9) years, (63.3 ± 3.1) mmHg (1 mmHg=0.133 kPa) vs. (51.8 ± 4.2) mmHg, 69.0％(29/42) vs. 39.9％(89/223) and 57.1％(24/42) vs. 25.1％(56/223), and the platelet was significantly lower than that in non-progression group:(168 ± 35) ×109/L vs. (189 ± 40) ×109/L, and there were statistical differences (P<0.05 or<0.01). The Logistic regression analysis result showed that renal dysfunction and CMBs were Independent risk factors of progressive CMBs (OR = 1.571 and 1.054, 95％ CI 1.042 - 2.493 and 1.010 - 1.142, P<0.05). Conclusions The rate of renal dysfunction is higher in patients of acute lacunar infarction with progressive CMBs, and progressive CMBs are associated with renal dysfunction.