Bloodletting Acupuncture at Jing-Well Points Alleviates Myocardial Injury in Acute Altitude Hypoxic Rats by Activating HIF-1α/BNIP3 Signaling-Mediated Mitochondrial Autophagy and Decreasing Oxidative Stress / 中国结合医学杂志

OBJECTIVE@#To explore the protective effect and possible mechanisms of bloodletting acupuncture at Jing-well points (BAJP) pre-treatment on acute hypobaric hypoxia (AHH)-induced myocardium injury rat.@*METHODS@#Seventy-five rats were randomly divided into 5 groups by a random number table: a control group (n=15), a model group (n=15), a BAJP group (n=15), a BAJP+3-methyladenine (3-MA) group (n=15), and a BANA (bloodletting at nonacupoint; tail bleeding, n=15) group. Except for the control group, the AHH rat model was established in the other groups, and the corresponding treatment methods were adopted. Enzyme-linked immunosorbent assay (ELISA) was used to detect creatine kinase isoenzyme MB (CK-MB) and cardiac troponins I (CTnI) levels in serum and superoxide dismutase (SOD) and malondialdehyde (MDA) levels in myocardial tissue. Hematoxylin-eosin (HE) staining was used to observe myocardial injury, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) staining was used to observe cell apoptosis. Transmission electron microscopy detection was used to observe mitochondrial damage and autophagosomes in the myocardium. The mitochondrial membrane potential of the myocardium was analyzed with the fluorescent dye JC-1. Mitochondrial respiratory chain complex (complex I, III, and IV) activities and ATPase in the myocardium were detected by mitochondrial respiratory chain complex assay kits. Western blot analysis was used to detect the autophagy index and hypoxia inducible factor-1α (HIF-1α)/Bcl-2 and adenovirus E1B 19k Da-interacting protein 3 (BNIP3) signaling.@*RESULTS@#BAJP reduced myocardial injury and inhibited myocardial cell apoptosis in AHH rats. BAJP pretreatment decreased MDA levels and increased SOD levels in AHH rats (all P<0.01). Moreover, BAJP pretreatment increased the mitochondrial membrane potential (P<0.01), mitochondrial respiratory chain complex (complexes I, III, and IV) activities (P<0.01), and mitochondrial ATPase activity in AHH rats (P<0.05). The results from electron microscopy demonstrated that BAJP pretreatment improved mitochondrial swelling and increased the autophagosome number in the myocardium of AHH rats. In addition, BAJP pretreatment activated the HIF-1α/BNIP3 pathway and autophagy. Finally, the results of using 3-MA to inhibit autophagy in BAJP-treated AHH rats showed that suppression of autophagy attenuated the treatment effects of BAJP in AHH rats, further proving that autophagy constitutes a potential target for BAJP treatment of AHH.@*CONCLUSION@#BAJP is an effective treatment for AHH-induced myocardial injury, and the mechanism might involve increasing HIF-1α/BNIP3 signaling-mediated autophagy and decreasing oxidative stress.

Effect of Tibetan medicine zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2 / 药学学报

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.

Effect of high altitude hypoxia on the activity and protein expression of CYP2C9 and CYP2C19 / 药学学报

This study is to investigate the effect of high altitude hypoxia on the activity and protein expression of CYP2C9 and CYP2C19. Rats from plain (P) and rats with acute middle altitude hypoxia (AMH), chronic middle altitude hypoxia (CMH), acute high altitude hypoxia (AHH) and chronic high altitude hypoxia (CHH) were administered orally phenytoin sodium (PHT) and omeprazole (OMZ) to evaluate the activity of CYP2C9 and CYP2C19, separately. The serum concentrations of PHT and metabolite 4'-hydroxyphenytoin (HPPH) at 12 h after treatment and the serum concentrations of OMZ and metabolite 5-hydroxy omeprazole (5-OHOMZ) at 3 h after treatment were determined by RP-HPLC. The activity of CYP2C9 and CYP2C19 was evaluated by the ratio of HPPH to PHT and the ratio of 5-OHOMZ to OMZ, respectively. The protein expressions of CYP2C9 and CYP2C19 were determined by ELISA method. The activities of CYP2C9 (HPPH/PHT) in P, AMH, CMH, AHH and CHH were 0.67 +/- 0.31, 0.75 +/- 0.29, 0.76 +/- 0.23, 0.79 +/- 0.31 and 0.75 +/- 0.18, respectively, and the activities of CYP2C19 (5-OHOMZ/OMZ) in P, AMH, CMH, AHH and CHH were 0.17 +/- 0.06, 0.20 +/- 0.10, 0.11 +/- 0.05, 0.37 +/- 0.13 and 0.19 +/- 0.05, respectively. The protein expressions of CYP2C9 in P, AMH, CMH, AHH and CHH were 4.20 +/- 1.27, 3.95 +/- 0.81, 3.93 +/- 1.11, 4.32 +/- 1.03 and 4.12 +/- 0.86 ng x g(-1), respectively, and the protein expressions of CYP2C19 in P, AMH, CMH, AHH and CHH were 3.91 +/- 1.82, 3.63 +/- 2.07, 2.55 +/- 0.85, 4.78 +/- 2.37 and 3.51 +/- 1.03 ng x g(-1), respectively. The activities and protein expressions of CYP2C9 in AMH, CMH, AHH and CHH were not significantly different with those of P. The protein expressions of CYP2C19 in AMH, CMH, AHH and CHH were not significantly different with those of P, but the activity of CYP2C19 in AHH was significantly higher than that of P. This study found significant changes in the activity of CYP2C19 under the special environment of acute high altitude hypoxia.

Autophagy of SO-Rb50 cells induced by arsenic trioxide / 中华实验眼科杂志

Background Cellular autophagy is a non-apoptosis death form of tumor tissue.Research determined that arsenie trioxide (As2O3) leads to apoptosis of tumor cells.But whether As2O3 induce autophagy of SO-Rb50 cells or not is unclear.Objective This study was to assess the effects of As2O3 on autophagy of SO-Rb50 cells.Methods As2O3 with the concentration of 0,0.5,1.0,2.0,4.0 μmol/L was used to treat the SO-Rb50 cell line for 48 hours,and the growth and proliferation of SO-Rb50 cells were detected using MTT assay (A570).pGFP-LC3,a marker of autophagy,was constructed to transfer SO-Rb50 cells,and the cells were then divided into RPMI-1640 culture group (untreated group),As2O3 + RPMI-1640 culture group (As2O3 treated group) and rapamycin culture group (positive control group).Autophagy of SO-Rb50 cells was examined by laser confocal microscope and monodansylcadaverine (MDC) influorescence staining,respectively,48 hours following cell culture.Ultrastructural features of autophagy were examined with transmission electron microscope (TEM).The percentage of autophagy positive cells in different concentrations of As2O3 treated groups was calculated with flow cytometer.Results The A570 values of SO-Rb50 cells were 2.194±0.066,1.841 ±0.213,1.035±0.046,0.374±0.042 and 0.167±0.019 in 0,0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups,with a significant difference among these 5 groups(F=547.636,P＜0.05),and those of 0.5,1.0,2.0,4.0 μmol/L As2O3 treated groups were significantly reduced in comparison with untreated group (P =0.000).The positive granular spots for GFP-LC3 chimeric protein were seen to aggregate in autophagic vacuoles in the As2O3 treated group and positive control group,but diffuse cytoplasmic signal for GFP-LC3 was found in the untreated group.Normal ultrastructure of SO-Rb50 cells was exhibited in the untreated group,and many double-membrane-like bound vesicles and autlysosomes were documented in the As2O3 treated group and positive control group under the TEM.A lots of MDC fluorescence granule were found in the As2O3 treated group and positive control group rather than the untreated group.Flow cytometry showed that the percentages of SO-Rb50 cells were 0,15.6％,42.7％,57.9％,79.5％ and 89.0％ in the 0,0.5,1.0,2.0,4.0 μmol/L As2O3 groups and positive control group,respectively,showing a As2O3 concentration-dependent increase.Conclusions As2O3 can induce the autophagy of SO-Rb50 cells and inhibit the proliferation of SO-Rb50 cells.Autophagic response of SO-Rb50 cells appears prior to the nuclear change after exposed to As2O3.The degree of autophagy of SO-Rb50 cells is associated with As2O3 dose.

A clinical and pathologic study of meibomian gland carcinoma with reduplicative operations / 中华实验眼科杂志

BackgroundThe meibomian gland carcinoma is an eyelid malignant tumor with a domestic incidence after basal cell carcinoma.Meibomian gland carcinoma is not sensitive to radiation therapy and chemotherapy,and the related factors with its recurrence and metastasis are rarely reported.ObjectiveThis study was to investigate the clinical and pathologic features of meibomian gland carcinoma with multiple operations and the effectiveness of histologically controlled excision.MethodsThe clinical data and the histopathologic sections of 34 cases of the meibomian gland carcinoma diagnosed by pathology were retrospectively analyzed at Zhongshan Ophthalmic Center in September 2003 to April 2011,and the treating effectiveness of histologically controlled excision was evaluated. ResultsIn this group of cases,the appearing rate of the meibomian gland carcinoma was resemble in both lateral eyes.A higher morbidity was on the upper eyelid (26/34,76.5％) and then the lower eyelids (5/34,14.7％ ) and both (3/34,8.8％).The average ages of these cases were 57.5 years old.Sixteen of 21 misdiagnosed cases were identified as chalazion at the first visit,and no histopathological examination was performed in 11 cases after initial operation.Twenty-six cases(76.5％ )were identified as meibomian gland carcinoma in initial histopathologic diagnosis.Two cases had histologically controlled excision and 16 cases had simple excision while 16 cases had chalazion enucleation in the first operation.All the patients had histologically controlled excision in Zhongshan Ophthalmic Center with 58.8％ of the patients having pagetoid invasion.Sixteen cases were followed up for 5 months to 8 years after histologically controlled excision,in which none died of recurrence and metastasis of meibomian gland carcinoma.No significant differences were found in the pathological feature between 16 lost patients and 18 followed-up patients(P ＞ 0.05 ).Conclusions Misdiagnosis of meibomian gland as chalazion is a main cause of repeat operations of meibomian gland carcinoma.Histologically controlled excision is a feasible therapy for the recurrence and metastasis of meibomian gland carcinoma.

Origin and development of human retinal astrocyte / 中华实验眼科杂志

BackgroundRetinal astrocytes are the main glial cells of retina,their origin and evolution progress are always the hot and difficult points in domestic and foreign researches. ObjectiveThis study was to explore the origin and development of retinal astrocytes in human fetal retina.Methods Thirty-three human embryonic eyes were collected from the abortion with the embryonic ages of 8-12 gestation weeks (20 eyes),15-17 gestation weeks ( 2 eyes),19-23 gestation weeks ( 4 eyes ),25 - 28 gestation weeks ( 4 eyes ),30 - 32 gestation weeks (3 eyes).The section of eyeball wall was prepared to observe the morphology and structure of different embryonic ages of retinas by regular histopathology examination.The origin of human embryonic retinal astrocytes was assessed by evaluating the change of GFAP expression in different embryonic ages of retinas using immunochemistry and immunofluorescence under the light and laser scanning confocal microscope.Theresearch was approved by the the Ethics Committee.Results The optic cup in embryonic 6- 7 weeks was in the retinal layering phase.Some immature short or round spindle-like cells appeared in primitive non-cell layer in the inner layer of the optic cup in embryonic 9 weeks of eyes.There were no positive GFAP-immunoactive cell was detected until embryonic 15 weeks of eyes.Some spindle-like cells migrated from a single layer primitive neuroepithelium next to optic disc expressing GFAP in the eyes with embryonic 19 weeks,and positive immunostaing for GFAP were detected in stellate cells surround blood vessels,and some seem to form the vessel wall in the ganglion cell layer and nerve fiber layer of the fetal central retina from 25 weeks through 26 weeks.Some positive response cells for GFAP presented in inner layer of the retina closed to ora serrata with the connection to nonpigmented epithelium (NPE) of the ciliary body during this period.Human retinal astrocytes showed typical stellate-like in shape,and the cellular processes crossed into inner plexiform layer in embryonic 28-week eyes.Conclusions Human retinal astrocytes mainly seem to have three kinds of origin in human embryonic eyes,and they are vascular precursor cell/pericytes,primitive neuroepithelium and nonpigmented epithelium of the ciliary body.

Application of avastin in vitrectomy for proliferative diabetic retinopathy and its mechanism / 中华实验眼科杂志

Background Researches demonstrated that avastin-assisted vitrectomy for serious proliferative diabetic retinopathy can decrease intra-operation complication and bring down difficulty of surgery.It is speculated that this is associated with suppression of avastin on neovascularization.However,the evidence of its pathology is lack.Objective The aim of this study was to evaluate the application and mechanism of avastin in vitrectomy for severe proliferative diabetic retinopathy. Methods Twenty-four eyes from consecutive 24 patients with vitrectomy for severe proliferative diabetic retinopathy were enrolled in this study.Fourteen eyes received all intravitreal injection of 0.06 ml avastin(1.5 mg)14 days prior to vitrectomy。And the other 10 eyes underwent only vitrectomy without avastin injection as controls,Preretinal membranes were collected during vitrectomy for histopathologic examination by hemotoxylin and eosin staining.The differences in the density of the neovessels and micro-neovessels between the two groups were observed by detecting the expression of CD34 in vesse]endothelial cells using immunohistochemistry.Written informed consent was obtained from each patient before surgery. Results No statistically significant differences were found in the demography and eye manifestations between only vitreetomy group and avastin+vitrectomy group(P＞0.05).The neovessels with grade three was in 10 eyes in only vitrectomy group and 1 eye in avastin+vitrectomy group(P＜0.01).More capillary-like neovascularization with single vascular endothelial cells,obvious hemorrhage and inflammatory cells infiltration were observed on preretinal membranes in vitrectomy group.However,there were less hemorrhage,ceUular components and few capillary-like neovascularization but more hyaline degeneration of fibrous tissue were observed in avastin+vitrectomy group under the light microscope.Immunochemistry revealed that CD34 was positively expressed in vascular endothelial cells on preretinal membrane in both two groups.The neovessels density and miero-neovessels density were 15.40±7.42/field and 1.88±1.70/field in avastin+vitrectomy group and those in vitreetomy group were 18.00±3.80/field and 0.45±0.56/field respectively,showing significant differences between these two groups(neovessels density:Z=-4.102,P＜0.01;micro-neovessels density:Z=-4.137,P＜0.01).Conclusion As an adjunct drug during the vitrectomy for proliferative diabetic retinopathy, avastin can improve the successful rate of surgery by inhibiting the neovascular formation and alleviating retinal edema.

Pharmacokinetics of sulfamethoxazole in healthy Han volunteers living at plain and in native Han and Tibetan healthy volunteers living at high altitude / 药学学报

The paper is to report the pharmacokinetics of sulfamethoxazole in healthy Han volunteers living at plain (PH) and native Han and Tibetan healthy volunteers living at high altitude (HNH and HNT). After healthy volunteers were administrated orally cotrimoxazole tablets, plasma concentration of sulfamethoxazole and metabolite N4-acetylsulfamethoxazole was determined by RP-HPLC, and plasma concentration-time data were analyzed by DAS 2.0 software to get the related pharmacokinetic parameters. The main pharmacokinetic parameters t(1/2) of sulfamethoxazole in PH, HNH and HNT were, respectively, 9.30 +/- 1.11, 10.99 +/- 1.23 and 10.44 +/- 1.05 h; tmax were 1.4 +/- 0.3, 2.0 +/- 1.1 and 1.8 +/- 0.4 h; Cmax were 94.42 +/- 15.26, 89.33 +/- 7.67 and 87.43 +/- 11.61 micro x mL(-1); AUC(0-t) were 1202.5 +/- 238.3, 1 434.7 +/- 193.9 and 1302.8 +/- 103.0 microg x h x mL(-1); AUC(0-infinity) were 1240.7 +/- 255.3, 1511.5 +/- 211.9 and 1363.9 +/- 116.5 microg x h x mL(-1); CL were 1.01 +/- 0.22, 0.81 +/- 0.12 and 0.89 +/- 0.08 L x h(-1) x kg(-1); V were 13.27 +/- 1.73, 12.81 +/- 2.15 and 13.28 +/- 1.20 L x kg(-1). Sulfamethoxazole pharmacokinetic parameters of HNH and HNT were significantly different from that of PH. The t(1/2) was significantly higher and the CL was significantly lower in HNH and HNT than that in PH, and the AUC(0-infinity) was significantly lower in HNT compared with HNH. This study found significant changes in the disposition of sulfamethoxazole under the special environment of high altitude hypoxia. This finding may provide some references for clinical rational application of sulfamethoxazole in HNH and HNT.

Pathological analysis of lacrimal pleomorphic adenoma / 中华实验眼科杂志

Background Lacrimal gland pleomorphic adenoma (LGPA) is the most common epithelialneoplasms of the lacrimal glands.Though its histopathological feature is benign,recurrence and malignant transition iscommon.The histopathologica feature of LGPA vary and resemble other lacrimal gland tumors.Objective Thissurvey was to explore the histopathological features of different subtypes of LGPA and their relationships with tumorrecurrence,and improve the knowledge of histopathological characteristics of LGPA.Methods A retrospectivestudy of 181 consecutive cases pathologically diagnosed as LGPA during 1966- 2010 years was performed toinvestigate the clinical and histopathological characteristics.Results One hundred and eighty-one LGPAs wereclassified as cell-rich type in 90(49.7％ ),stroma-rich type in 38 cases(21％ ),and intermediate type in 53 cases(29.3％ ).91.7％ of all tumors showed areas with thin( ＜20 μm) capsules independent of the tumor subtype.Tumorsof stroma-rich subtype showed greater thin-capsule regions.86.5％ recurrent LGPAs belonged to stroma-rich subtype(x2 =120.896,P =0.000 ),which had a higher rate of pseudopodia or satellite nodules in the capsules than other twosubtypes did(x2 =80.715,P =0.000 ).Cuboidal cells were the most commonly found cellular type followed bysquamous cells.Duct-like structures were the most frequent patterns formed mainly by epithelial cells.Myxoid stromawas the most frequently found mesenchymal-like tissue.Conclusions The tendency of recurrence and invadationof capsule may contributed to the high recurrence rate in stroma-rich type of LGPAs.The knowledge of the immensevariety of cells,architectures and morphological characteristics of LGPA are essential for a correction of pathologicaldiagnosis.

Study on genetic aberrations of ocular mucosa-associated lymphoid tissue lymphomas occurring in southern China / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the genetic aberrations of ocular extranodal marginal zone B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type occurring in patients from southern China.</p><p><b>METHODS</b>Fifty seven paraffin-embedded ocular MALT lymphoma specimens from patients in southern China were studied by interphase fluorescence-in-situ hybridization (FISH) for genetic aberrations including t(11;18)(q21;q21)/API2-MALT1, t(1;14)(p22;q32)/IgH-bcl-10, t(14;18) (q32;q21)/IgH-MALT1 and bcl-6/FOXP1 gene translocations.</p><p><b>RESULTS</b>Amongst the 57 cases studied, 9 cases (15.8%) showed chromosome translocations, including 4 cases (7.0%) of t(11;18)(q21;q21)/API2-MALT1, 1 case (1.8%) of t(14;18) (q32;q21)/IgH-MALT1, 1 case (1.8%) of bcl-6 gene-related chromosome translocation and 3 cases (5.3%) of IgH-unknown translocation partner. FISH revealed 17 cases (29.8%) with 3 copies of bcl-6 gene, 21 cases (36.8%) with 3 copies of MALT1 gene and 12 cases (21.1%) with 3 copies of both genes.</p><p><b>CONCLUSIONS</b>The MALT lymphoma-associated chromosome translocations t(11;18)(q21;q21)/API2-MALT1 and t(14;18) (q32;q21)/IgH-MALT1 are demonstrated in ocular MALT lymphomas of southern Chinese patients. The prevalence is significantly different from that reported in northern Chinese and northern American patients, indicating a geographic heterogeneity in the MALT lymphoma-associated genetic aberrations. The presence of 3 copies of bcl-6 and MALT1 genes is the commonest genetic abnormalities observed in ocular MALT lymphomas, suggesting a possible role in MALT lymphomagenesis.</p>

Multi-points pulse information acquisition and 3-D reconstruction based on grid-net images / 中国医疗器械杂志

A method of acquiring radial pulse information from the grid-net area on every pulse image frame is proposed based on the principle of lens imaging and the characteristics of image data. The radial pulse image data are collected by the pulse image sensor. Multi-points out-of-plane displacements of the pulse are acquired by the method. And the membrane surfaces are reconstructed, then the 3D information of the pulse can be observable.

Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of retinoblastoma / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To find new biomarkers in the sera of retinoblastoma (Rb) patients with surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI TOF MS) and protein chip technique.METHODS:SELDI TOF MS, IMAC30 and CM10 protein chips were used to analyze the protein profiles from sera of 18 patients with Rb and 17 age matched controls. The protein profiling was analyzed statistically by Ciphergen protein chip software 3.0.2. The test was applied to compare the protein peak intensity. Fisher's exact test was used to compare the predominance of differential protein peaks appeared in patients.RESULTS:With IMAC30 protein chips, there were 26 proteins which appeared different in sera of patients with Rb compared to normal children. Among them, 21 proteins, I.e. 7746, 7014, 11713, 3049, 7084, 7299, 5888, 2544, 12575, 5489, 9658, 9575, 9929, 10161, 8955, 1886, 10617, 6209, 2411, 7374, 6614m/z were up regulated and 5 proteins, I.e. 8382, 7923, 7972, 8590, 66576m/z, were down regulated(P<0.01). Using the 7014 protein peak for statistical analysis, we could differentiate the patients with Rb from the healthy children with a sensitivity of 94.4% and a specificity of 82.4%. By CM10 protein chips, 4 proteins, including 3 up regulated proteins(5888, 6097, 7798 and 1 down regulated protein (8590m/z), were detected in Rb patients (P<0.01). The sensitivity and specificity were 83.3% and 70.6% respectively when 7798m/z protein peak was selected for statistical analysis.CONCLUSION:There are a few candidates as Rb biomarkers in the sera of Rb patients. SELDI TOF MS protein chip technology could be a potential method in the clinical screening test of Rb.