Objective To construct recombinant lentivirus silence vector aiming at rictor gene in mTORC2 specific protein, and to investigate its regulation on mTORC2/SGK1 signal pathway and the effect on pulmonary alveolar epithelial sodium ion chan-nel,as well as the role in acute respiratory distress syndrome(ARDS)and acute lung injury.Methods The interfering vector plas-mid and empty vector plasmid of target gene rictor were constructed,which and the lentivirus packaging system were co-transfected to 293T cells.The viral supernatant was collected,centrifuged,concentrated and purified for obtaining recombinant lentivirus.The virus titer was detected and the virus was infected to A549 cells.Stable cell lines were screened.RT-PCR was used to confirm the silencing situation of target gene rictor.The expression situation of various signal indexes in this pathway was detected by PCR and Western blot.Results The recombinant lentivirus of silence gene rictor was successfully constructed and transfected to A549 cell for obtaining stable cell lines.Compared with blank and control groups,the mRNA levels of rictor,downstream SGK1 andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased (P <0.05 ).Meanwhile,the protein levels of rictor,downstream SGK1,P-SGK andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased compared with the other two groups(P<0.05).Conclusion Silence rictor gene has the obvious regulation effect on mTORC2/SGK1 signal pathway,meanwhile affects the expression of pulmonary alveolar epithelial cellular α-,β-and γ-ENaC at gene and protein level.It is speculated that mTORC2/SGK1 may be an important signal pathway for regulating the clearance capacity of pulmonary alveolar epithelial cells on pulmonary alveolar fluid and simultaneously affecting the pulmonary edema formation.

Objective To explore the effects of balloon dilatation on esophageal motility in patients with achalasia. Methods Forty eight patients were diagnosed as having achalasia by clinical features, barium radiography, endoscopy and esophageal manometry (PC polygraf HR, Medtronic). The following parameters were evaluated: the symptom score, the maximal width of the esophagus (MWE) on barium radiography, lower esophageal sphincter pressure (LESP), lower esophageal sphincter relaxation rate (LESRR) and the contraction amplitude of esophageal body before and after dilatation at 4 weeks and 12～24 weeks. Results (1) Parameters of symptom score and the maximal esophagus width were significantly decreased ( P

Calcium calculi account for about 80% of urolithiasis with high incidence and recurrence rate. Recurrent urolithiasis increases the risk of hypertension and chronic kidney disease. Comprehensive metabolic evaluation is of paramount importance for the prevention of calcium calculi. With deepening of the study on the pathogenesis of calculi, we find crystallization inhibitors, inflammation and oxidative stress molecules play an important role in the formation of calculi, and susceptibility gene sites of urinary calculi are identified in succession. These may be used as novel markers for the prevention and evaluation of urinary calculi. Models that predict the risk of kidney stone recurrence can help warn people at high risk.

AIM: To dynamically observe the feeling change of the photorecrptor layer in the eyes with acute central serous chorioretinopathy ( CSCR ) krypton laser treatment by fourier - domain optical coherence tomography ( FD - OCT), and to study their correlation with the chang of vision.
METHODS: This is a retrospective case series study. The clinical diagnosis of 52 patients with monocular initial onset of central serous chorioretinopathy, krypton laser photocoagulation before treatment, after 1, 2, 4, 6, 8wk, 6mo, FD - OCT were performed to observe the morphological changes characteristic of photoreceptor layer and changes in vision.
RESULTS: After 1wk treatment, all cases were improved; 2wk, 6 cases were cured; 4wk, 38 cases were cured; 6wk, 41 cases were cured; 8wk, 45 cases were cured, the OCT showed macular retinal neuroepithelial layer ( RNL ) from fully absorbed; 6mo with the same 8wk. Before and after treatment in patients with best corrected visual acuity and from the height difference between the macular region of RNL was statistically significant (P<0. 05), there was a correlation between the changes of visual acuity after treatment and the macular detachment of RNL height (P<0. 05), Photoreceptor layer of complete and incomplete best corrected visual acuity difference was statistically significant (P<0. 01).
CONCLUSION: FD-OCT can dynamicaly observed acute central serous chorioretinopathy krypton laser treatment of photoreceptor ultrastruture changes. Photoreceptor layer of complete and incomplete best corrected visual acuity difference was statistically significant (P<0. 01).

Objective: To explore the relationship between prevalence of atrial fibrillation (AF), iskhemia stoke and CHA2DS2-VASc score in patients≥65 years in order to provide prevention and treatment basis in clinical practice. Methods: A total of 5016 patients admitted in our hospital from 2013-10 to 2015-10 were enrolled. The patients were divided into 2 groups: AF group, n=437 and Non-AF patients, n=4579; according to age, the patients were further assigned into 4 subgroups as <65 years subgroup, (65-74) years subgroup, (75-84) years subgroup and ≥85 years subgroup. The risk factors for AF occurrence were retrospectively studied. Results: Compared with the Non-AF group, the patients in AF group had the elder age and more male gender, both P<0.001; more patients combining with hypertension, coronary artery disease (CAD), diabetes, sick sinus syndrome and rheumatic heart disease, all P<0.001. Age, male gender, CAD, sick sinus syndrome and rheumatic heart disease were the independent risk factors for AF occurrence. Compared with Non-AF group, AF group showed the higher prevalence rate of ischemic stroke and the elder onset age, both P<0.01. For non-valvular AF, the ratio of patients with CHA2DS2-VASc score≥2 was higher than those with CHA2DS2-VASc score<2 and the rate of anticoagulant therapy was decreasing by age increasing, all P<0.001. Conclusion: Age, male gender, CAD, sick sinus syndrome and rheumatic heart disease were independently related to AF occurrence. Non-valvular AF patients had the higher risk for ischemic stroke than non-AF patients, anticoagulation therapy should be conducted at the early stage.

Objective To study inflammatory cytokines changes in brain tissue of neonatal rats with periventricular leukomalacia (PVL).Method A total of 80 neonatal SD rats (P3) were randomly assigned into 2 equal groups,sham-operated group and PVL group.Rats in each group were further assigned into four subgroups (12,24,48,72 h),with 10 rats in each subgroup.The hypoxic-ischemic PVL modal were established following the procedure:first,isolation and ligation of left common carotid artery,and then exposed to 8％ O2 and 92％ N2 for 2.5 h.The sham-operated rats were processed with isolation of left common carotid artery only.Rats of the four subgroups were sacrificed at 12,24,48 h and 72 h respectively,then the brains were rapidly removed in corresponding time.Pathological changes of brain tissues were observed using HE stain.The mRNA expression levels of tumor necrosis factor α(TNF-α)and inter leukin-1β (IL-1β) were assessed using real-time quantitative PCR assays,the protein levels of TNF-α and IL-1β were detected using enzyme linked immunosorbent assay method.Result The brains tissues of rats in PVL group showed remarkably hyperemia and edema,with left ventricle enlargement.Periventricular white matter structure was disintegrated comparing with sham-operated group.The expression of TNF-α and IL-1 β mRNA in PVL group increased significantly,reaching peak by 24 h and then gradually decreased 72 h after the procedure.The mRNA levels of TNF-c and IL-1 β were significantly different between each two time points of 12,24 h and 48 h in PVL group (P ＜0.05).However,there was no differences between 72 h and 48 h within PVL and sham-operated group group(P ＞0.05).In PVL group,the protein expression trends of TNF-α and IL-1 β were similar to mRNA expression trends.Moreover,the protein levels were significantly different between each two time points of TNF-α and IL-1 β,respectively (P ＜ 0.01).The protein expression levels of TNF-α were different at each time point between PVL group and sham-operated group[(189.2 ± 20.4) pg/ml vs.(131.4 ±5.2) pg/ml at 12 h,(213.8 ± 16.7) pg/ml vs.(127.7 ±7.4) pg/ml at 24 h,(181.7 ± 15.0) pg/ml vs.(126.3 ± 6.0) pg/ml at 48 h,(159.6 ± 25.3) pg/ml vs.(131.4 ± 6.0) pg/ml at 72 h;P ＜0.01].The protein levels of IL-1β were different between the two groups only at 24 h and 48 h.[(121.8 ±30.0) pg/ml vs.(67.4 ± 13.7) pg/ml,(83.3 ± 15.7) pg/ml vs.(65.3 ± 14.9) pg/ml;P ＜0.05].In sham-operated group,no differences of TNF-α and IL-1 β protein levels were found between any different time points (P ＞ 0.05).Conclusion Inflammatory cytokines such as TNF-α and IL-1β are involved in ischemic-hypoxia induced PVL.Dynamic detection of inflammatory factors is expected to be an important method of early diagnosis,assessment of treatment efficacy and prognosis of PVL.

Objective To evaluate the effect of olmesartan medoxomil tablets (olmesartan) in comparison with Olmetec on 24 h ambulatory blood pressure (ABPM) and blood pressure variability (BPV)in patients with mild to moderate hypertension.Methods A randomized,double-blind,double-mimic controlled trial was performed.Forty-eight patients with mild to moderate essential hypertension were randomly into treatment group (olmesartan) and control group (Olmetec) for eight weeks.The ABPM was taken before and at the end of the trial.Results After eight weeks,treatment with olmesartan induced a significant reduction in ABPM in patients [(9 ± 3)/(11 ± 3) mmHg (1 mmHg =0.133 kPa)],which is similar with the reduction by Olmetec [(9 ± 4) / (9 ± 5) mmHg],P ＞ 0.05.This situation holds for BPV with the standard deviations of 24 h,systolic blood pressure/diastolic blood pressure of pre-treatment and pro-treatment were (10 ± 2)/(11 ± 3) mmHg vs (10 ± 3)/(12 ± 2) mmHg in olmesartan group,and (10 ± 3)/(11 ±3) mmHg vs (12 ±3)/(12 ±4) mmHg in Olmetec group.(3) There is no difference in the rate of adverse event between olmesartan (10.42％) and Olmetec (8.33％) treatment(P ＞ 0.05).Conclusion Similar to Olmetec,treatment with olmesartan once daily can significantly reduce ABPM in patients with mild to moderate essential hypertension.

Background Studies showed that some members of matrix metalloproteinases (MMPs) play an important role in the pathogenesis of diabetic retinopathy (DR).However,whether MMPs inhibitor can deter blood retinal barrier (BRB) from damage is below understood.Objective This study was to investigate the effects of GM6001,a MMPs inhibitor,on BRB permeability.Methods Twenty-four adult clean SD rats were randomized to the control group,diabetic group and diabetes+GM6001 group according to randomized number table.Diabetic models were induced by the intraperitoneal injection of streptozotocin in the rats of the diabetic group and the diabetes + GM6001 group,and equal volume of citrate buffer was used in the same way in the rats of the control group.GM6001 10 μ1 (100 μ mol/L) was intravitreously injected in the third and fourteenth day after modeling in the diabetes+ GM6001 group,and equal volume of normal saline solution was injected in the same way in the control group and the diabetic group.The rats were sacrificed and eyeballs were extracted 1 month after injection,and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in rat retinas were detected by reverse transcription PCR (RT-PCR).Evens blue (EB) was infused via the right jugular vein,and paraformaldehyde solution 1％ was then infused via left ventricle at the perfusion pressure 120 mmHg.The eyeballs were extracted 2 minutes later,and the leakage of EB in rat retinas was examined.Results RT-PCR electrophoresis exhibited the response bands of MMP-2 mRNA,and MMP-9 mRNA and GAPDH,with the gene size of 436,536 and 484 bp,respectively.The difference of the MMP2 mRNA and MMP-9 mRNA was statisticaly significant (F =20.336,P =0.000 ; F =8.742,P =0.002) ; and the relative expressions of MMP-2 mRNA and MMP-9 mRNA were significantly higher in the diabetic group and diabetes +GM6001 group than those in the control group (all at P＜0.01),and the relative expressions of MMP-2 mRNA and MMP-9 mRNA in the diabetes+GM6001 group was significantly reduced in comparison with the diabetic group(both P=0.01,P=0.02).The standardized EB content in the retinas of the control group,diabetic group and diabetes+ GM6001 group was (12.60±3.50) ng/mg,(26.52±7.14) ng/mg and (17.55±2.65) ng/mg,showing a significant difference (F=17.032,P＜0.01),and EB content in rat retinas in the diabetic group was higher than that in the control group (P=0.003),and that in the diabetes+GM6001 group was lower in comparison with the diabetic group (P=0.020).Conclusions Intravitreal injection of GM6001 can down-regulate the expression of MMP-2 mRNA and MMP-9 mRNA in diabetic rats and therefore protect BRB.

Objective Human sperm hyaluronidase in the acrosome is a key enzyme during fertilization.To study and establish a modified substrate film method to improve the diagnotic and treatment of male infertility and investigate the influence of sperm hyaluronidase on male fertility.Methods According to the biochemical feature of sperm hyaluronidase that can dissolve the hyaluronic acid in matrixes,the modified sodium hyaluronate-Gelatin membrane was used as substrate to demonstrate the sperm hyaluronidase activity by incubation and staining.70 human semen samples were selected,categorized and detected for hyaluronidase activity according to the resuh of routine clinical semen examination.The average sperm hyaluronidase activity was statistically anyliezed between the fertile group and infertile group.Results Under general light microscope,the clear and bright digestion circle around the sperm head can be observed against the background of deep purple-blue stained substrate at the positive-reaction area.The amounts of positive reaction and the diameter of the bright circle are positively related to the activity of sperm hyaluronidase.The average hyaluronidase reaction rate and diameter of normal fertile group were 70.84% and 78.17 ?m;What about the infertile group A were 60.02% and 76.92 ?m;The infertile group B were 29.11% and 8.22 ?m; There was statistical difference of HYD activity between the infertile group.B and the fertile group(P

The neurotoxin β-amyloid (Aβ) is the main hallmark of Alzheimer′s disease (AD). Recent evidence suggests that, in common sporadic or late-onset forms of AD, elevated brain Aβ levels are caused by impaired clearance rather than overproduction. The cell surface receptor′s low-density lipoprotein receptor-related protein-1 (LRP1) has been reported to not only play a role in Aβ endocytosis, but also exist in the blood-brain barrier system, peripheral blood, liver, kidney and other tissues and organs, and transport Aβ to the cerebrospinal fluid or blood system by passing through the blood-brain barrier or the blood-cerebrospinal fluid barrier effectively, and finally clear it out of the body through peripheral tissues and organs. In this review, the role of LRP1 in the peripheral transport and clearance of Aβ is described, and it may be a safe and effective way to reduce Aβ in the brain and even improve cognitive dysfunction.