Objective To investigate the value of distinguishing small clear cell renal cell carcinoma and high-attenuation renal cysts by applying unenhanced CT value and homogeneity of the esions.Methods Retrospective analysis of 38 cases of small renal cell carcinoma confirmed by operation and pathology and 58 cases of high-attenuation renal cyst that were confirmed by enhanced CT,and measured CT value and homogeneity of each mass,and did statistical analysis for corresponding datas.Results The CT value of small clear cell renal cell carcinoma was (35.5±10.1)HU (range 20-60.3 HU).The CT value of high-attenuation renal cysts was (70±1 5.2)HU (range 29-95 HU).Difference of the two groups had statistical significance (P <0.001).For high-attenuation re-nal cysts,uniform density often can be seen(47 cases,81.03%).For small clear cell renal cell carcinoma,uneven density often can be seen (29 cases,76.32%).Difference of the two groups had statistical significance (P <0.001).When optimal cut-off of CT value determined by ROC was 5 1.5 HU,the sensitivity and specificity were 89.7% and 92.1%,respectively.Conclusion Unenhanced CT may be helpful in distinguishing small clear cell renal cell carcinoma and high-attenuation renal cysts,which can provide reasona-ble suggestions on the next identification examinations.

Objective To investigate the efficacy and safety of tiotropium bromide aerosol combined with salmeterol in the treatment of ACOS. Methods 76 patients with ACOS were enrolled in this study from October 2014 to November 2016. They were divided into experimental group and control group according to the random number table method. The test group was given tiotropium bromide combined with salmeterol Casson aerosol treatment, the control group was given salmeterol tacrolone aerosol treatment, compared the two groups of patients with clinical efficacy and adverse reactions. Results The total effective rate was 92.11% in the experimental group and 73.68% in the control group, the difference was statistically significant (Z = 4.547, P<0.05). The ACT score of the test group was higher than that of the control group, the CAT score was lower than that of the control group, the difference was statistically significant (P<0.05); The incidence of adverse reactions in the test group was 10.53%, the incidence of adverse reactions in the control group was 13.16%,the difference was not statistically significant (χ2 = 0.126). Conclusion Tiotropium powder combined with salmeterol tegon aerosol in the treatment of bronchial asthma-chronic obstructive pulmonary syndrome is reliable and worthy of clinical practice.

Background Phakic posterior chamber implantable contact lens (PPC-ICL) or phakic posterior chamber Toric implantable contact lens (PPC-TICL) implantation is an effective way for the correction of high myopia or high myopia with astigmia,but it often has residual myopic power.Excimer laser-assisted subepithelial keratectomy (LASEK) can correct the residual myopia following PPC-ICL or PPC-TICL,but its effectiveness and safety deserve attention.Objective This study was to analyze the clinical effectiveness and safety of LASEK for residual myopia after PPC-ICL implantation for extreme high myopia.Methods A prospective cases-observational study was performed,and written informed consent was obtained from each patient before any surgery.Fourteen eyes of 9 patients with residual myopia following PPC-ICL or PPC-TICL for the eyes with spherical equivalent refraction of ≥-20.00 D were collected in the Affiliated Hospital of Guizhou Medical University from July 2010 to March 2015,including PPCICL implantation in 8 eyes and PPC-TICL implantation in 6 eyes.LASEK were performed on the eyes to correct the residual myopic power.Uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),haze,the distance of intraocular lens to lens,corneal thickness,corneal topography,corneal endothelial cell counting,intraocular pressure (IOP) and fundus were examined and compared before and after surgery.The effectiveness and safety of the surgery were evaluated.Results The operation was smooth and no complication was found after surgery in all of the eyes.The UCVA and BCVA were significantly different in the eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F =31.360,1.778;both at P＜0.05),and the UCVA after LASEK was higher than BCVA before LASEK.The refractive powers were (-22.27-±4.29),(-3.75±2.25) and (-0.42±0.63) D before surgery,6 months after PPC-ICL implantation and 12 months after LASEK,showing a significant difference among them (F=46.370,P＜0.05),and the refractive power was considerably lower after LASEK than that before surgery and after PPC-ICL implantation (both at P＜0.05).No significant difference was found in IOP or corneal endothelial cell counting in operated eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F=1.663,1.055;both at P＞0.05).The distance of intraocular lens to lens was (0.69±0.26)mm in the eyes after LASEK and (0.71 ±0.29)mm in the eyes after PPC-ICL implantation,with no significant difference between them (t =0.192,P＞0.05).Conclusions PPC-ICL or PPC-TICL implantation for the correction extreme high myopia often remains a certain degree of myopia,and LASEK for the correction of residual refractive power is safe and effective.

Aim To investigate the roles of p53-dependent signaling pathways in the process of ginsenoside Rg1 protection against 8-methoxypsoralen(8-MOP) and subsequent ultraviolet A(UVA) irradiation induced photoaging model in human dermal fibroblasts(HDFs).Methods Photoaging model was established by 8-MOP/UVA in skin HDFs.Flow cytometry, enzyme cytochemistry, immunofluorescence and Western blot were employed.Results Pretreatment with ginsenoside Rg1 could significantly reduce the amount of UVA-generated 8-oxo-dG and relieve the photoaging representation.Compared with 8-MOP/UVA treatment group, pretreatment with Ginsenoside Rg1 could decrease the expression of SA β-galatosides(SA-β-Gal), and down-regulate the level of senescence associated proteins(p16 and p21).Conclusions Ginsenoside Rg1 has prominent dose-dependent antagonism on senescence of skin HDFs induced by 8-MOP/UVA, and its mechanism may be due to its antioxidation which reduces the production of photoproducts to protect telomere against abnormal shortening.

Objective To evaluate the effect of early-phase insulin secretion and insulin resistance in the pathogenesis of type 2 diabetes, and to analysis the risk factors of glucose tolerance deterioration. Methods Oral glucose tolerance test (OGTT) was performed in subjects over 30 years old coming from 78 families with type 2 diabetes. A total of 118 subjects with normal glucose tolerance (NGT) [fasting plasma glucose (FPG)<6.1 mmol/L and 2h postprandial glucose (2hPG)<7.8 mmol/L] were enrolled. Another OGTT was performed in them to define the glucose tolerance status at the end of the 4-7 years follow-up. AINS30/APG30, the ratio of the increment of insulin to that of plasma glucose at 30 min after the glucose load, was used to assess the early phase insulin secretion. HOMA-IR and HOMA-β were calculated to assess the insulin resistance and β-cell function respectively. Results After 4-7 years follow-up, 66 of 118 subjects still remained NGT, while 52 became either diabetic (n=11)or pre-diabetic (n=41). Using the median of HOMA-IR and AINS30/APG30 as the cutoff points, all subjects were divided into four groups: subjects with good early phase insulin secretion and no insulin resistance, subjects with good early insulin secretion but relative insulin resistance, subjects with impaired early phase insulin secretion but no insulin resistance, subjects with impaired early phase insulin secretion and also relative insulin resistance. The incidences of abnormal glucose tolerance among these four groups were 23.1%, 36.4%, 45.5% and 73.1% respectively. There was a statistical difference between the former three groups and the last one (P<0.05). Log/st/c regression analysis showed that only the early phase insulin secretion was the risk factor of glucose tolerance deterioration, while age, gender, insulin resistance or β-cell function were not. Conclusion Impaired early phase insulin secretion is a major risk factor for the disturbance of glucose metabolism in the population with NGT.

Hepatitis B virus (HBV) DNA replication takes place in the viral capsid that consists of 180 or 240 copies of HBV capsid (HBc or core) protein. The monomeric core protein contains an apical loop region that forms the spikes on the surface of viral capsid upon core dimerization and capsid assembly. To investigate the impact on HBV DNA replication through gene engineering at the spike of HBV capsid. plasmids expressing engineered HBc with linker-fused enhanced green fluorescent protein (EGFP) or shortened EGFP insertion at the spike region were constructed by Restriction Digestion and Ligation-independent Cloning (RLIC). The wildtype or mutant HBc construct was cotransfected with HBV1.1c(-), a plasmid containing 1.1 unit-length HBV genome with deficiency in HBc expression, into HEK293 cells, respectively. GFP signal was observed through a fluorescence microscope and HBV DNA replicative intermediates were assayed by Southern blotting to determine the expression and functions of different recombinants. Our results demonstrated that the RLIC method was effective to generate deletion or insertion in the apical loop region of HBc. Both HBc-EGFP recombinants with different linkers produced green fluorescence but with different subcellular distribution pattern. However, HBV DNA replication was not detected with the trans-complementation of these two HBc recombinants. In addition, other recombinants including the one only with the deletion of aa79-80 failed to support HBV replication. Taken together, our results suggest that RLIC is a robust method which can be broadly applied in gene engineering; different peptide linkers may have different influences on the functions of an engineered fusion protein; and HBc aa79-80 play a critical role for HBc to support HBV DNA replication.
Capsid Proteins ; genetics ; Cloning, Molecular ; Genetic Engineering ; methods ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection ; Virus Replication

Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.

Objective To prepare Integrin αvβ3/CCR2 dual targeted microbubble,test physical-chemical properties,enhancement effect and targeting ability in vitro.Methods The dual targeted microbubbles (MBdual-target )with FITC labled iRGD and PE labled CCR2 were prapared,and non-target microbubbles as control (MBcontrol ) were prapered.Physical and chemical properties of two groups of microbubbles were tested,connectivity of peptides/antibodies and microbubbles were detected by fluorescence microscope and flow cytometry instrument.Enhancement effect and the stability of two groups of microbubbles was observed and compared in vitro.The affinity of MBdual-target and MBcontrol for bEnd.3 cells was investigated with light and fluorescent microscope.Results ①The particle size of MBdual-target was (0.93±0.23)μm,with no statistically significant difference compared with MBcontrol (P >0.05).②MBcontrol showed no fluorescent,while MBdual-target showed both clear green and red light,under fluorescent microscope.③There was no significant difference of gray scale of enhancement between MBdual-target and MBcontrol in vitro.④ It was showed that MBdual-target adhered to bEnd.3 cells in vitro experiment.Conclusions Integrinαvβ3/CCR2 dual targeted targeted microbubbles was successfully prepared and proved having good enhancement effect and targeting ability in vitro.

Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.

Objective To study the relationship between serum cystatin C(Cys C)level and the development of insulin resistance(IR)in elderly patients with type 2 diabetes mellitus (T2DM).Methods A cross-sectional survey research involving 757 elderly patients with T2DM was performed and patients were divided into groups according to Cys C and IR levels.Serum 1evels of fasting insulin (FBI),fasting blood glucose(FPG),fasting C-peptide (FCP),glycosylated hemoglobin A1 c (H bA1 c),homeostasis model assessment of insulin resistance (IR) (HOMA2 IR-C),homeostasis model assessment of insulin secretion,homeostasis model assessment of insulin sensitivity(HOMA2-％ S-C),micro-albuminuria(mALB)and serum lipid were measured and compared among the groups.Possible influence factors were adjusted,and the correlation between Cys C levels and IR was analyzed.The influencing factors on IR were also analyzed.Results There were statistically significant differences in ages,course of T2 DM,FBI,FCP,HbA1c,urinary mALB,IR,insulin secretion,insulin sensitivity,morbidity rate of coronary heart disease,hypertension and diabetic nephropathy among the three groups(allP＜0.05).Insulin sensitivity was decreased with the increase of Cys C level,while others were increased.Among 757 patients,the level of serum Cys C was positively correlated with FCP,HOMA2-IR C levels,and was negatively correlated with HOMA2-％ S-C levels(P＜0.05).Multiple logistic regression analysis showed that the higher level of Cys C was independent risk factors for IR in elderly T2DM patients(OR=3.41,95％CI:2.22-5.22,P＜0.05).Conclusions Serum Cys C level is closely related with IR in elderly T2DM,and the elevated level of serum Cys C is one of independent risk factors for elderly T2DM.