Objective To investigate the effect of melatonin on oxidative stress and inflammatory reaction in patients with moderate to severe chronic obstructive pulmonary disease and to explore its mechanisms. Methods 42 patients with moderate to severity chronic obstructive pulmonary disease in stable stage were random-ly divided into melatonin group and control group,and 21 patients in each group treated with melatonin(3 mg/d) or placebo for 3 months respectively. The plasma levels of 8- isoprotane,IL-8,TNF-α,h-CRP pulmonary func-tion,six minutes walking test and MRC dyspnea score before treatment,2 months and 3 months after the treatment were analyzed. Results After 2 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(10.40 ± 5.4 vs. 16.92 ± 4.33,P<0.05),and the concentration of IL-8 (6.88 ± 2.37 vs. 11.33 ± 3.39,P < 0.05). After 3 months of treatment,compared to placebo groups,melatonin could significantly decrease the concentration of 8-isoprotane(9.40 ± 4.0 vs. 17.92 ± 3.33,P < 0.01),and IL-8 (5.67 ± 3.22 vs. 9.31 ± 3.23,P < 0.05). Compared with before treatment,melatonin could significantly de-creased the concentration of 8-isoprotane(9.40 ± 4.0 vs. 20.40 ± 8.4,P<0.01 )and IL-8(5.67 ± 3.22 vs. 12.33 ± 3.88,P<0.05)after 3 months. Meanwhile,the concentration of the TNF-α(25.83 ± 9.18 vs. 35.83 ± 12.18,P<0.05)and hypersensitive C(1.76 ± 1.18 vs. 3.09 ± 1.79,P < 0.05)reactive protein in the melatonin group was greatly lower than the placebo group. After 3 months,compared to the placebo group,MRC dyspnea score of pa-tients in the group of melatonin was improved significantly(1.56 ± 1.38 vs. 2.09 ± 1.16,P<0.05 ),and lung func-tion and six minutes walk test showed no significant difference between patients in the two groups. Conclusions Exogenous melatonin administration can decrease the concentration of 8-isoprotane,IL-8,TNF-αand h-CRP in the blood of patients with moderate to severe COPD ,and improve the MRC dyspnea score. Melatonin has a significant effect on reducing oxidative stress and inhibiting inflammatory reaction in patients with moderate and severe stage stable COPD,which demonstrates its potential therapeutic value with broad clinical application prospects.

OBJECTIVE:To establish a method for the determination of related substances in bifonazole raw material. METH-ODS:HPLC method was performed on the column of Kromasil C18 with mobile phase of methanol-0.02 mol/L phosphoric acid(ad-justed pH to 7.5 with triethylamine)(70:30,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 258 nm,volume injection was 20 μl,and the column temperature was 40 ℃. RESULTS:The linear range was 0.05-0.25 μg/ml for bifonazole(r=0.9996), 0.05-0.25 μg/ml for impurity A(bifonol)(r=0.9997)and 0.05-0.25 μg/ml for impurity B(4-C isomer)(r=0.9995);the detec-tion limits were 8.2 ng/ml,7.5 ng/ml and 8.4 ng/ml,and the quantification limits were 27.1 ng/ml,24.7 ng/ml and 27.8 ng/ml,re-spectively;RSDs of precision and reproducibility tests were lower than 1%;recovery of impurity B was 95.13%-101.29%(RSD=1.89%,n=9);both impurity A and impurity B were were detected in the 3 batches of samples. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the determination of the related substances in bifonazole raw material.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.

Stroke is one of the severe perioperative complications.The incidence of perioperative stroke has great difference because of the surgical site,surgical complexity,and different basic diseases of patients.The induced anesthesia and the intraoperative management also have an important influence on the onset of stroke.The mortality of perioperative stroke is as high as 26%.Neturologists must guide the assessment of the potential risks for perioperative stroke,and carefully balance the risk-benefit ratio when making each decision.This article reviews the advances in research on the risk factors for perioperative stroke,prevention,and treatment.

Objective To improve the understanding to transposition of the great arteries with posterior aorta and decrease the clinic misdiagnosis by summarizing their echocardiography features . Methods Echocardiography images were retrospectively reviewed for 7 cases with transposition of the great arteries with posterior aorta who diagnosed in Anzhen Hospital ,and their echocardiography features were summarized .Results In 7 patients ,5 patients had operation ,including 3 for arterial switch operation ,1 for right GLEEN ,1 for right GLEEN and Fortan surgery ,and 2 patients were lost to follow‐up .The echocardiography characters:① Artery short view showed aorta and pulmonary artery existed the half‐surrounding relationship .The aorta was situated posteriorly and the pulmonary artery anteriorly .②Multiple views showed aorta was completely and mostly arised from the right ventricle ,while the pulmonary artery was came off the left ventricle .③ Ventricular septal defects were presented in all cases and aorta could ride the defect of ventricular septal .④ In most cases ,fibrous continuity between the aortic and mitral valve through a ventricular septal defect was found and there was no conus under the aorta .⑤ Some cases could combine the different degree stenosis of the outflow tract of subpulmonary valve and pulmonary valve . Conclusions The connection relationship between two arteries and ventricles can displayed clearly on the long axis view of double outflow tracts of two ventricles under xiphoid process .The acoustic window under the xiphoid process is the key for diagnosis and differential diagnosis of the disease .

Objective To improve the prenatal echocardiographic diagnostic accuracy for fetus with isolated mirror-image right aortic arch which combined with vascular ring by analyzing and accumulating its echocardiographic features.Methods Echocardiographic signs were analyzed retrospectively and fetal echocardiographic features were accumulated in 16 cases with prenatal diagnosis of isolated mirror-image right aortic arch with left-side ductus arteriosus which formed vascular ring between January 2014 and February 2016.All cases were confirmed by neonatal echocardiography.Results The fetal echocardiography characters:① In 3-vessel and trachea view,fetal right aortic arch was demonstrated a Ushaped appearance of the great vessels,the right arm of the ‘ U’ represented the aortic arch and its left branch with ductal continuation.② In 3-vessel and trachea view or upper axial mediastinal transverse view,left innominate artery passed straightly to the left of the trachea.Left innominate artery and right-side ductus arteriosus were overlapped and the distance of two vessels was very close.③ In double arch view and longitudinal view for ductus,Left innominate artery did not connect to descending aorta or ductus arteriosus and did not involve in the formation of vascular ring.④ Color Doppler played an important role in diagnosis of this disease.Power Doppler or high-definition imaging easily showed color overflow and wrongly displayed the crossed relationship of left innominate artery with descending aorta as the anatomical connection,and caused the misdiagnosis of this disease as double aortic arch.Conclusions Isolated mirrorimage right aortic arch with vascular ring is not rare in the fetal period.Because ductus arteriosus is large and patent during the prenatal period,it is easily mistaken for the crossed relationship of left innominate artery with descending aorta as the anatomical connection and finally misdiagnoses this anomaly as double aortic arch.The keys to accurate diagnosis of the disease are clearly visualization of the innominate artery and its relationship with descending aorta and ductus arteriosus by scanning different views.

AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

Background Diabetic retinopathy (DR) is one of the leading causes that result in adult irreversible blindness in many countries.Recent researches suggest that neurodegeneration is an important component of DR.To realize the disease process of retinal neutron is very important for prevention and treatment on DR.Objective This study was to investigate the change of retinal nerve fiber layer thickness in patients with type 2 diabetes mellitus.Methods Ninety-six eyes of 48 patients with type 2 diabetes mellitus were enrolled in Peking Union Medical College Hospital.The patients were assigned into non-diabetic retinopathy (NDR) group,background diabetic retinopathy(BDR) group,proliferative diabetic retinopathy (PDR) group and panretinal photocoagulation (PRP) group based on the fundus finding and fundus fluorescein angiography(FFA),and 24 normal subjects with matched age were included as control group.RNFL thickness was measured by GDxVCC system,including temporal,superior,nasal,inferior,total,(TSNIT) average,superior average,inferior average,TSNIT standard deviation and nerve fiber indication.The datas of the RNFL thickness were analyzed and comparison among different groups by one-way analysis of variance and Student Newman Keuls test.Results The TSNIT averages of the NDR group,BDR group,PDR group and PRP group were(56.54±5.28),(56.92±6.49),(53.04±6.14) and(53.17±9.30) μm,respectively,while that of the control group was (59.04±4.37) μm.The TSNIT average,superior average,inferior average,TSNIT standard deviation of the PDR group and PRP group compared with control group were significantly decreased,and the nerve fiber indication of the PDR group and PRP group was significantly increased (P =0.002,0.000,0.002,0.000,0.001 ;P =0.002,0.000,0.001,0.000,0.000).Compared with the control group,the TSNIT average,superior average,inferior average,TSNIT standard deviation were insignificantly decreased,and the nerve fiber indication was insignificantly increased in the NDR group and BDR group (P =0.187,0.235,0.333,0.106,0.202 ;P=0.262,0.063,0.072,0.098,0.062).Conclusions The decline of the RNFL thickness appears prior to DR findings.The RNFL thinning of PDR and PRP patients suggests the degeneration of neurons and atrophy of axonal.The neurodegeneration is an important component of DR.

Objective To detect infection of Angiostrongylus cantonensis and examine effection of treatment to prepare monoclonal antibodies(McAbs). Methods Six-week-old BALB/c mice were imrnunized by the intraperitoneal injection of e/s antigens of Angiostrongylus cantonensis. Fusion of splecn cells from immunized mice with prepared SP2/0-Ag14 myeloma cells was performed in RPMI 1640. Fused cells were suspended in RPMI 1640 containing 1% HAT and 20% fetal calf serum and dispensed into 96-well cell culture plates. The supernatants of clones were screened by ELISA with sera of patients of angiostrongyliasis.Distribution of cohere antigen of 12D5 and 21B7 monoclonal antibodies was analyzed with immunohistochemistry. Two McAbs ( 12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA.Results 12D5 McAb was identified as IgG1 and 21 B7 McAb was IgM. Western blot result showed two McAbs could used to identified 55 × 103 protein of adult worms of A. cantonensis. Cohere antigen of 12D5 and 21B7 monoclonal antibodies were distributed on intestine surface of A. cantonensis. The detection rates of CAg in the sera of infected rats 100% (48/48), the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostomiasis, trichinosis, anisakiasis as well as schsitosomiasis, and health srea did not reacted with 12D5 and 21B7 McAbs,and detaction rate of antibody of angiostrongyliasis patients only reached 75% (24/32) with antigen of A. cantonensis. Conclusion Cohere antigen of 12D5 and 21B7monoclonal antibodies were antigens of enteric epithelium. Sandwich ELISA with 12D5 and 21B7 McAbs showed high specificity act as detecting CAg of A. cantonensis in sera of infection animal and patients. It is apparent that Sandwich ELISA with 12D5 and 21 B7 is not only rapid and simple without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A. cantonensis.