Objective To make objective criteria for evaluation of conjunctival impression cytology(CIC)in early screening of vitamin A deficiency(VAD)in children.Methods The status of vitamin A in children aged 3～7 years old was evaluated with CIC method.In order to justify the judgemental criterion of CIC,the changes in CIC and the levels of serum vitamin A were studied.Results Goblet cells and normal epithelial cells could be found in grade 0～1 of CIC;only normal epithelial cells without Goblet cells could be seen in grade 2 of CIC,and only abnormal epithelial cells were identified in grade 3～5 of CIC.Along with the increasing degree of abnormalify of CIC,the levels of serum vitamin A decreased accordingly(r=-0.824,P

Objective:To investigate the effect and mechanism of transcription factor Foxp 3 on the proliferation and cell cycle of human lung adenocarcinoma cell line A 549.Methods:We knocked down the expression of Foxp 3 using siRNA.Foxp3 inhibition was detected by RT-PCR.Cell proliferation was detected by MTT.Cell cycle of A549 cells were detected by flow cytometry after the transfection of siRNA.Cell cycle-related checkpoint genes were filtered by RT-PCR.The regulation of Foxp3 on cell cycle-related checkpoint genes were detected by immunofluorescence and dual -luciferase reporter assay system.Results: The proliferation of A549 cells were inhibited after silencing Foxp3,and A549 cells were arrested in G0/G1 cycle.G1/S cycle checkpoint gene CCND1 was down regulated.Mechanism research show that Foxp 3 can regulate the expression of CCND 1 directly.Conclusion: Foxp3 can promote the proliferation of A549 cell line by up regulating G 1/S cycle checkpoint gene CCND 1.This provides a new target for the therapeutic targets of lung adenocarcinoma.

Objective:To study whether autoimmune regulator (Aire) affects macrophage polarization. Methods: The mouse mononuclear macrophage cell line RAW264. 7 cells, stable expressing GFP-Aire protein RAW264. 7 cells ( A33-3 ) and stable expressing GFP protein RAW264. 7 cells (C1-6) were stimulated with LPS,IL-4 and LPS combined with immune complex respectively to make the macrophage polarize to M1 (LPS),M2a (IL-4) and M2b (LPS with immune complex). To investigate the effects of Aire on the various types of macrophages polarization,the M1 related molecules (IL-1α,iNOS and IL-6),M2a related molecules (Arg-1) and M2b related molecule (IL-10) were detected by Real-time PCR. Results:The expression of IL-1α,IL-6 and inducible nitric oxide synthase ( iNOS) ,the products of M1 macrophages, were significantly upregulated in RAW264. 7 cells treated with LPS at 0. 5 μg/ml. The expression of Arg-1 and IL-10 mRNA in RAW264. 7 cells increased in a dose-dependent manner after stimulation with IL-4 or LPS combined with immune complexes, respectively. M1 macrophage-related marker iNOS and IL-1 increased, whereas IL-6 levels decreased in A33-3 cells compared with C1-6 cells after treatment with LPS. The expression of Arg1 and IL-10 were downregulated in A33-3 cells compared with C1-6 cells after IL-4 and LPS combined with immune complexes stimulation,respectively. Conclusion:Aire may promote M1 macrophage polarization while inhibit M2a and M2b macrophage polarization.

Objective To investigate clinical results of the treatment of posterior shoulder instability in a combined way of bone block procedure and posterior capsulorrhaphy. Methods The double contrast CT arthrography was performed for 5 patients suffering from posterior shoulder instability so as to detect the pathology. The treatments combining bone block procedure and posterior capsulorrhaphy were carried out according to the pathological conditions of bone and soft tissue. Results The double contrast CT arthrography showed posterior bone defects, posterior glenoid labrum tear and enlargement of posterior capsular cavity. After the bone block procedure and posterior capsulorrhaphy, no patients complained shoulder instability in the follow-up of 5 to 38 months. Conclusions Many pathological factors may induce posterior shoulder instability, which should be corrected by an appropriate operation rather than a “standard”method of operation. For patients involving several pathological factors, a combined operation may be needed in order to correct various pathological changes.

Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.

Resveratrol is a natural phytoalexin with special pharmacological and health functions. Stilbene synthase (STS) is a key and rate-limiting enzyme in the biosynthesis of resveratrol that is present only in a limited number of plants. The content of resveratrol from Polygonum cuspidatum is more than 1000 times higher than grapes and peanuts. We speculate that the catalytic ability of different STS may be one of the reasons causing differences in the content of resveratrol. To verify the above speculation, Vitis vinifera stilbene synthase gene (VvSTS) was amplified according to overlap PCR protocol with genomic DNA as template. VvSTS and PcSTS (PcPKS5) were analyzed through heterologous expression in Escherichia coli. The expression products were purified with Ni-NTA sepharose affinity chromatography and desalted through PD-10 column. The molecular weight of the two fusion proteins was about 43 kDa. Enzyme reaction and product analysis showed that the two products were resveratrol. The enzyme kinetic analysis showed that the catalyze efficiency (Kcat/Km) of PcPKS5 was 2.4 times of the VvSTS. Our findings confirms that STS from certain plants has much higher catalytic capability.
Acyltransferases ; metabolism ; Fallopia japonica ; enzymology ; Recombinant Fusion Proteins ; biosynthesis ; Stilbenes ; metabolism ; Vitis ; enzymology

To investigate the effect of captopril on the dentin bonding durability of self-etch adhesive. Different concentrations of captopril ethanol solutions or captopril ethanol/water solutions were prepared to pretreat dentin as primer for the self-etch adhesives. The surface morphology of the dentin was observed with scanning electron microscopy (SEM). Based on the morphology analysis, the pretreatment condition was selected and two self-etch adhesives were employed to evaluate the improvement effect of the captopril pretreatment on the dentin bonding durability. : SEM showed that the pretreatment of captopril ethanol solutions and captopril ethanol/water solutions were able to remove the smear lay and partially expose collagen matrix. According to the SEM results, the pretreating condition of captopril ethanol/water solution with the pretreating time of was selected for further dentin bonding study. For Clearfil SEBOND system, the immediate bonding strength increased from to  (<0.05). After one-year aging, the bonding strength of the control group decreased markedly [(22.90±6.82) MPa, <0.05]; while the bonding strength of the captopril pretreated group kept steadily >0.05]. For Clearfil S BOND system, there was no significant difference in the immediate bonding strength between the experimental group [(4.07) MPa] and the control group[(4.11) MPa]. But after one-year aging, the bonding strength of the experimental group was higher than that of the control group <0.05]. : The pretreatment with captopril ethanol/water solution increases the dentin bonding strength of the self-etch adhesive systems and also improves the bonding durability.
Adhesives ; Captopril ; Dental Bonding ; Dentin ; Dentin-Bonding Agents ; Materials Testing ; Microscopy, Electron, Scanning ; Resin Cements

OBJECTIVETo explore the feasibility of inducing fungal infection by Cryptococcus neoformans aerosol inhalation in immunosuppressed Balb/c mice.

METHODSTwenty-four Balb/c mice were randomized into cyclophosphamide (CTX) group and control group (n=12). The mice in CTX group were subject to inhalation of Cryptococcus neoformans aerosol prepared using a ultrasonic nebulizer 4 days after intraperitoneal CTX injection, and the control mice received the inhalation only. The leukocyte count and changes in appetite, body weight, and behaviors of the mice were observed after the treatments. On days 3 and 7 after the inoculation, the mice were sacrificed to analyze the fungal counts in brain and lung tissue homogenates and examine the pulmonary pathologies.

RESULTSCTX injection caused lowered appetite and body weight loss in the mice, and significantly reduced the leukocyte counts (2.77∓0.45 vs 8.26∓0.56, P<0.05). At days 3 and 7 after inoculation, the Cryptococci load in the lungs increased obviously in CTX group with a colony-forming unit (CFU) of the lungs of 271.67∓122.22 and 41.67∓0.28, respectively, significantly higher than that in the control group (60.00∓43.36 and 3.00∓5.30, respectively, P<0.05). In CTX group, the CFU was 10.17∓5.42 and 9.17∓6.34 in the peripheral blood and 6.83∓4.92 and 11.00∓5.44 in the brain tissue at days 3 and 7, respectively, whereas no Cryptococci was detected in the peripheral blood or in the brain tissue in the control group. Pathological examination of the lungs revealed destruction of normal alveolar structure in CTX group, with numerous infiltrating inflammatory cells and visible yeast.

CONCLUSIONInhalation of Cryptococcus neoformans aerosol can cause fungal infection in the lungs of immunosuppressed Balb/c mice.

Aerosols ; Animals ; Brain ; microbiology ; Colony Count, Microbial ; Cryptococcosis ; microbiology ; Cryptococcus neoformans ; growth & development ; Disease Models, Animal ; Immunocompromised Host ; Inhalation ; Lung Diseases, Fungal ; microbiology ; Male ; Mice ; Mice, Inbred BALB C ; Ultrasonics

Objective:To investigate the preventive and therapeutic effects of resveratrol on lens opacification in diabetic rats and its biological mechanism.Methods:Fifty 8-week-old healthy male SPF grade SD rats were selected and randomly divided into blank control group, model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group according to their body weight, with 10 rats in each group.The diabetes model was established by intraperitoneal injection of streptozotocin in model group, gliclazide group, low-dose resveratrol group and high-dose resveratrol group.On the third day after modeling, rats in gliclazide group was gavaged with 2 mg/(kg·d) gliclazide suspension, and rats in low-dose and high-dose resveratrol groups were gavaged with 20 and 40 mg/(kg·d) resveratrol, respectively, for four weeks.Rats in blank control group and model group were gavaged with the same volume of normal saline once a day, also for four weeks.After the diabetes model was established, there were 10 rats in blank control group and 9 rats in the other four groups.The fasting blood glucose concentration of the rats was measured with a blood glucose meter.The concentrations of fasting insulin, superoxide dismutase (SOD) 1, SOD2, SOD3, and glutathione peroxidase (GPX1) were determined by enzyme-linked immunosorbent assay.Lens opacification after treatment was observed by slit lamp microscopy.Morphologic changes in lens cells were examined by hematoxylin-eosin staining.Apoptosis of lens epithelial cells (LECs) was detected using TUNEL.The relative expressions of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in lens tissues were determined by Western blot.The study protocol was approved by the Welfare Ethics Committee of Experimental Animal of Zhengzhou University (No.IACYC2019-02).Results:Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were increased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were decreased in model group in comparison with blank control group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group, low-dose resveratrol group, and high-dose resveratrol group compared with model group, and the differences were statistically significant (all at P<0.05). Fasting blood glucose concentration, fasting insulin level, and apoptosis rate of LECs were decreased and the concentrations of SOD1, SOD2, SOD3, and GPX1 were increased in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, and the differences were statistically significant (all at P<0.05). The proportions of grade 0, 1 and 2 lens opacities after treatment were 100.00%, 0.00% and 0.00% in blank control group, 0.00%, 66.67% and 33.33% in model group, 77.78%, 22.22% and 0.00% in gliclazide group, 22.22%, 44.44% and 33.33% in low-dose resveratrol group, and 66.67%, 33.33% and 0.00% in high-dose resveratrol group, respectively, with a statistically significant difference ( H=7.514, P<0.001). Compared with model group, lens opacification was less severe in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). Lens opacification was less severe in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Compared with model group, there were fewer abnormal changes of lens cells and sub-organelles in gliclazide group, low-dose resveratrol group and high-dose resveratrol group, and the abnormalities in gliclazide group and high-dose resveratrol group were slighter.Compared with model group, the relative expression levels of Nrf2 and HO-1 were higher in blank control group, gliclazide group, low-dose resveratrol group, and high-dose resveratrol group, with statistically significant differences (all at P<0.05). The relative expression levels of Nrf2 and HO-1 were higher in gliclazide group and high-dose resveratrol group compared with low-dose resveratrol group, showing statistically significant differences (both at P<0.05). Conclusions:Resveratrol can reduce lens opacification in diabetic rats and its mechanism may be related to the regulation of the Nrf2/HO-1 signaling pathway by exerting antioxidative stress effects.

Objective: To develop a native adaptive behavior scale for children with autism spectrum disorder(ASD) and to explore its reliability and validity. Methods: Items of ASD adaptive behavior rating scale were selected based on the scale development theory, ASD knowledge and adaptive behavior concept through preliminary survey and statistical, and 301 ASD children aged 2 to 12 from hospitals in Guangzhou, Huizhou, Shenzhen who met the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition were selected, data was analyzed by the item analysis. Results: After item analysis and exploratory factor analysis, the final version of the scale contains 58 items, and 64.24% of the total variation could be explained by 6 factors; The Cronbach’s α coefficient of the full scale was 0.98, and the coefficient value of dimen sional factors were 0.94，0.93，0.91，0.95，0.88，0.94. The test-test reliability r of full scale was 0.86, the r of the factor were 0.88，0.81，0.81，0.87，0.88，0.79. The criterion-related validity r with the ABAS-Ⅱ scale was -0.77, the criterion-related validity r with the CARS scale was 0.64. Conclusion The ASD Child Adaptive Behavior Scale showed good reliability and validity, and could be used widely.