Objective To investigate the change in heat shock protein 70(HSP7O) in the brain tissue of fetal and newborn rats after prenatal hypoxic adaptation and the possible mechanism of the protective effect. Methods Twenty-two 22d pregnant Wistar rats were randomly divided into two groups: GroupⅠ(hypoxic adaptation group) and group Ⅱ(control group). The animals in group Ⅰ were placed in a tightly closed hypoxic adaptation chamber, of which the oxygen and carbon dioxide concentrations were monitored. The pregnant rats were taken out and exposed to fresh air for 5 mm when the 02 % in the chamber was reduced to 15 %, then the pregnant rats were placed back in the chamber and the above process was repeated once. The animals were then left for their natural labor. In control group the pregnant rats underwent the same process but the chamber was not tightly closed(O2 %= 21 %). Prenatal rats were delivered by cesarean section at lb. 3h, 8h, 24h, 48h, 72h, 120h and 168h after hypoxic adaptation and decapitated and brain was removed. Seven newborn rats from each group were decapitated and brain was removed for determination of HSP70 expression with immunohistochimical technique. Results No HSP7O expression was observed in the brain tissue of normal prenatal and newborn rats. HSP70 was observed in the different regions of hippocampus and cortex from 8h to 168h after hypoxic adaptation. Strongest HSP70 expression was observed in hippocampus CAl . Conclusions HSP70 plays a role in the formation of prenatal hypoxic adaptation.

AIM:To compare the effect of three types of pterygium surgery and on tear film in patients with type 2 diabetes mellitus. ·METHODS:A total of 102 patients ( 102 eyes ) with pterygium combined with type 2 diabetes mellitus treated in our hospital from March 2013 to March 2016 were analyzed retrospectively. The patients were divided into three groups including the 34 cases ( 34 eyes ) with simple excision of pterygium ( resection group ) , pterygium excision combined with conjunctival flap transplantation in 34 cases (34 eyes, as conjunctival flap group ) and pterygium excision combined with limbal stem cell transplantation in 34 cases ( 34 eyes, as stem cell group ) . The wound repair time, complications, recurrence rate, uncorrected visual acuity (UCVA), tear film break-up time ( BUT ) and basal tear secretion test (SⅠt) were observed before, and 6 and 12mo after surgery in the three groups, respectively. ·RESULTS:The postoperative UCVA of the three groups was significantly higher than that preoperation ( P =0. 039, 0. 013, 0. 024 ), and there was no significant difference among the three groups ( P = 0. 317 ). The wound repair time was 5. 67 ± 1. 45d in the resection group, which was significantly higher than that in the conjunctival flap group (4. 18 ± 0. 76d) and the stem cell group (4. 09±0. 79 d) (P<0. 001), there was no significant difference between the conjunctival flap group and the stem cell group ( P = 0. 937 ). There were 4 cases in resection group reappeared, and the recurrence rate was 11. 8%, which was significantly higher than the other two groups ( P = 0. 037 ). There were 1 recurrences in the conjunctival flap group, and the recurrence rate was 2. 9%, while the patients in the stem cell group had no obvious recurrence. SⅠt and BUT increased significantly after operation (P<0. 05), especially in conjunctival flap group and stem cell group (P<0. 001). There was no significant difference between the conjunctival flap group and the stem cell group (P=0. 845, 0. 894). · CONCLUSION: Pterygium excision combined with conjunctival flap transplantation or limbal stem cell transplantation for the treatment of type 2 diabetic patients with normal blood glucose and tear film function has the similar effect, and is better than simple pterygium excision.

BackgroundThe effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood.ObjectiveThe present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.MethodsHFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining.Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased.ConclusionsELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.

Objective To establish a REDE-DHPLC method for detecting the EGFR and KRAS mutations in plasma DNA from tumor patients, and investigate its clinical significance. Methods Restriction endonucleases Mse Ⅰ , Msc Ⅰ , BstN Ⅰ and Bgl Ⅰ were used to digest the wild type fragments of exon 19,exon 21 of EGFR gene and coden 12, 13 of KRAS gene for enriching the mutation fragments, and REDE-DHPLC method was established to detect EGFR and KRAS mutations. The sensitivities of REDE-DHPLC and conventional DHPLC were analyzed by using a series of plasmids containing 50%, 10%, 5%, 1% and 0. 1% mutation genes. Then, Plasma samples and paraffin-embedded tissue samples of 120 NSCLC patients and 120 colorectal cancer patients were detected by REDE-DHPLC. Compared with conventional DHPLC and sequencing, the diagnostic efficiency of REDE-DHPLC method was evaluated by detecting the mutation status of 2 genes in plasma of NSCLC and colorectal cancer patients. Results The sensitivity values of REDE-DHPLC and conventional DHPLC for detecting mutations in 4 loci were 0. 1% and 1%respectively. Plasmid DNA containing 0.1% mutation gene was detected to be positive continually for 2 to 3 times by REDE-DHPLC. EGFR mutation rates of 120 plasma from NSCLC patients detected by REDE-DHPLC, conventional DHPLC and sequencing methods were 27. 5%, 16. 7% and 12.5% respectively, and KRAS mutation rates of 120 plasma from colorectal cancer patients were 38. 3%, 25. 8% and 16. 7%,respectively. The positive rates of EGFR and KRAS mutation detected by REDE-DHPLC were significantly higher than conventional DHPLC(x2 = 4. 092, 4. 301, all P ＜ 0. 05 ) and sequencing method (x2= 8. 438,14. 127,all P ＜ 0. 05 ). In comparison with conventional DHPLC, the sensitivities of REDE-DHPLC for detecting EGFR and KRAS mutation were 100% (20/20,31/31), the specificities were 87. 0% (87/100)and 83. 2% (74/89). In comparison with sequencing method, the sensitivities of REDE-DHPLC were 100%( 15/15,20/20), the specificities were 82.9% (87/105)and 74. 0% (74/100). The coincidence rate of the two methods for detecting EGFR and KRAS mutation were 89. 2% ( 107/120, Kappa = 0. 690, P ＜ 0. 05 ) and 87.5% ( 105/120, Kappa= 0. 718, P ＜ 0. 05 ). The Consistency of EGFR and KRAS mutation status in plasma and tissues detected by REDE-DHPLC were 91.7% (33/36, Kappa =0. 939,P ＜0. 05)and 90. 2 %(46/51, Kappa = 0. 914, P ＜ 0. 05 ), respectively. Conclusions The REDE-DHPLC method is highly sensitive and specific for detecting EGFR and KRAS mutations in plasma DNA from tumor patients. The results are easy to be interpreted without missing homozygous point mutation, which indicate that the detection of EGFR and KRAS mutations in plasma DNA by REDE-DHPLC could therefore extend to be usedin clinical laboratory.

Objective To explore the protection of valsartan combined with simvastatin on kidney in early diabetic nephropathy rats. Methods Diabetic nephropathy rats model were induced by streptozocin (STZ) ,the experimental rats were randomly divided into 5 groups: control (group C), diabetic nephropathy (group D) ,diabetes treated with valsartan (group X) ,diabetes treated with simvastatin (group Z) ,and diabetes treated with combined valsartan and simvastatin ( group L). Blood glucose (BG), HbA1c, blood cholesterol ( TC), trigalloylglycerol ( TG ), blood ureanitrogen ( BUN ), serum creatinine (SCr) , urinary albumin excretion rate (UAER) were measured, and the podocyte ultrastructure was observed by transmission electronic microscopy. Results The levels of BG, HbA1c,TC,TG and UAER in group D increased significantly compared togroup C(BG:[20.3 ±3.2]mmol/L vs [6.1 -±0. 4]mmol/L;HbA1c:[7.18 ±0.47]% vs [3.37 ±0. 15]% ;TC: [2. 69 ±0. 35] mmol/L vs [1.28 ±0. 24] mmol/L;TG: [3.09 ±0. 37] mmol/L vs [1.18 ±0. 25]mmol/L) (P ＜ 0. 05 ). Creatinine clearance rates (Ccr) in group D ( [0. 89 ± 0. 19] ml/min ) decreased significantly compared to group C( [1.27 ±0. 33] ml/min) ,as well as group X,Z and L( Ps ＜ 0. 05 ). UAER in group D was significantly higher than that in group C ( [19. 87 ±3. 85] μg/24 h vs [3. 67 ± 1.01] μg/24 h) (P ＜ 0. 05 ), as well as group X, Z and L ( P ＜ 0. 05 ), and the improvement in group L was particularly significant ( P ＜ 0. 05 ). The projections of podocyte in group D severely syncretized, there were slightly improvement in group X, Z and L compared to group D, and the improvement in group L was remarkable. Conclusion The treatment with valsartan, simvastatin and their combination will effectively protect the kidney in early diabetic nephropathy rats,and the effect of using the combination therapy is much better.

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the effect of KyoT2 on the proliferation and migration of airway smooth muscle cells (ASMCs) in mice with asthma.

METHODSOvalbumin (OVA) was used to establish the asthmatic model of airway remodeling in BALB/c mice. ASMCs were isolated and cultured, and primarily cultured ASMCs were used as the control group. The expression of KyoT2 in ASMCs was measured in the control and asthma groups. After the ASMCs from asthmatic mice were transfected with pCMV-Myc (empty vector group) or pCMV-Myc-KyoT2 plasmid with overexpressed KyoT2 (KyoT2 expression group) for 48 hours, RT-PCR and Western blot were used to measure the mRNA and protein expression of KyoT2, the MTT assay and BrdU assay were used to measure the proliferation of ASMCs, and Transwell assay was used to measure the migration of ASMCs. Western blot was used to determine the effect of KyoT2 overexpression on the protein expression of RBP-Jκ, PTEN, and AKT.

RESULTSCompared with the control group, the asthma group had significantly downregulated expression of KyoT2 in ASMCs, and the KyoT2 expression group had significantly upregulated expression of KyoT2 in ASMCs (P<0.05). Compared with the empty vector group, overexpressed KyoT2 significantly inhibited cell proliferation and migration, downregulated the expression of RBP-Jκ and AKT, and upregulated the expression of PTEN.

CONCLUSIONSOverexpressed KyoT2 can inhibit the proliferation and migration of ASMCs through the negative regulation of RBP-Jκ/PTEN/AKT signaling pathway.

Animals ; Asthma ; pathology ; Cell Movement ; Cell Proliferation ; Female ; Intracellular Signaling Peptides and Proteins ; physiology ; LIM Domain Proteins ; physiology ; Mice ; Mice, Inbred BALB C ; Muscle Proteins ; physiology ; Myocytes, Smooth Muscle ; physiology ; PTEN Phosphohydrolase ; physiology ; Trachea ; pathology

Age Factors ; Aged ; Female ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Scrub Typhus ; diagnosis ; pathology

Objective The aim of this study was to analyze the status and influencing factors of public health service utilization in rural patients with severe mental illness in Yunnan province. Methods The main caregivers of patients with severe mental illness in Yunnan province were selected by stratified cluster sampling method. Multivariate logistic regression was used to analyze the influencing factors of public health service utilization. Results A total of 284 cases of rural patients with severe mental illness were investigated, including 144 males (50.7％) and 140 females (49.3％) . The rate of medical file filing, follow-up rate of village doctor, participation rate of free physical examination and health education acceptance rate among patients with severe mental illness were 89.8％, 84.9％, 73.2％and 56.7％respectively (<0.05) . Female patients had higher participation rate of free physical examination than male patients, and patients having work and patients with controlled disease condition and their caregivers had higher rates of health education acceptance rate than their counterparts ( <0.05) . Conclusion Measures should be taken to strengthen the work of medical examination for male patients, and to expand health education activities for patients having no work and patients with uncontrolled disease condition, so as to improve the public health service utilization in rural areas of Yunnan province.

Neuropeptide Y (NPY) co-exists with norepinephrine (NE) in sympathetic terminals, and is the most abundant neuropeptide in myocardium. Many studies have focused on the effects of NE on ion channels in cardiac myocytes and its physiological significance has been elucidated relatively profoundly. There have been few investigations, however, on the physiological significance of NPY in myocardium. The effects of NPY on L-type Ca2+ channel currents (I(Ca-L)) were evaluated in some studies and different results were presented, which might be attributed to the different species of animal tested and different methods used. It is necessary, therefore, to study the effects of NPY on ion channels in cardiac myocytes systematically and further to discuss the biological significance of their coexistence with NE in sympathetic terminals. The single ventricular myocytes from adult rat or guinea pig (only for measuring I(K)) were prepared using enzymatic dispersion. I(Ca-L), I(to), I(Na/Ca), I(Na) and I(K) in the cellular membrane were observed using whole cell voltage-clamp recording. In the present study, NPY from 1.0 to 100 nmol/L dose-dependently inhibited I(Ca-L) (P<0.01, n=5). The maximal rate of inhibition in this study reached 39% and IC(50) was 1.86 nmol/L. NPY had no effect on the voltage-dependence of calcium current amplitude and on the voltage-dependence of the steady-state gating variables. I(Ca-L) was activated at -30 mV, reaching the maximum at 0 mV. When both NE and NPY were applied with a concentration ratio of 500:1, 10 nmol/L NPY inhibited I(Ca-L) that had been increased by 5 mumol/L NE, which was consistent with the effect of NPY only on I(Ca-L). NPY also inhibited I(Na/Ca). At a concentration of 10 nmol/L, NPY inhibited inward and outward I(Na/Ca) from (0.27+/-0.11) pA/pF and (0.45+/-0.12) pA/pF to (0.06+/-0.01) pA/pF and (0.27+/-0.09) pA/pF, respectively (P<0.05, n=4). NPY at 10 nmol/L increased I(to) from (12.5+/-0.70) pA/pF to (14.7+/-0.59) pA/pF(P<0.05, n=4). NPY at 10 nmol/L did not affect I(Na) in rat myocytes and I(K) in guinea pig myocytes. NPY increased the speed of action potential depolarization and reduced action potential duration of I(Ca-L), I(Na/Ca) and I(to), which contributed to the reduction of contraction. These results indicate that the effects of NPY are opposite to the effects of NE on ion channels of cardiac myocytes.
Animals ; Calcium Channel Blockers ; pharmacology ; Calcium Channels, L-Type ; drug effects ; Female ; Guinea Pigs ; Heart Ventricles ; cytology ; Ion Channels ; drug effects ; Male ; Myocytes, Cardiac ; metabolism ; Neuropeptide Y ; pharmacology ; Norepinephrine ; Patch-Clamp Techniques ; Rats ; Rats, Wistar ; Sodium-Calcium Exchanger ; antagonists & inhibitors