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OBJECTIVE@#To investigate the recovery of protective effects of exogenous hydrogen sulfide (HS) on hypoxia post-conditioning in aged H9C2 cells and its mechanism.@*METHODS@#H9C2 cells (cardiomyocytes line) were treated with 30 μmol/L hydrogen peroxide (HO) for 2 hours, then cultured for 3 days in order to induce cellular aging. Aged H9C2 cells were randomly divided into 5 groups (=8):Control group (Control), hypoxia/reoxygenation group (H/R), H/R + NaHS group, hypoxia post-conditioning (PC) group, PC+NaHS group. H/R model:the cells were exposed to hypoxic culture medium (serum and sugar free medium, pH=6.8) for 3 hours and then cultured at normal condition for 6 hours. PC model:at the end of hypoxia for 3 hours, the cells were exposed to normoxic culture solution for 5 minutes, then the cells were placed in hypoxic solution for 5 minutes, the cycle above-mentioned was repeated 3 times and followed by reoxygenation for 6 hours. Advanced glycation end products (AGEs) content and caspase-3 activity were detected by ELISA. The cell viability was observed by cell counting kit-8 (CCK-8). The reactive oxygen species (ROS) levels were analyzed using 2, 7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. The apoptotic rate was determined through Hoechst 33342 staining. The mRNA levels of relative gene expression were detected by real-time PCR.@*RESULTS@#Thirty μmol/L HO induced H9C2 cell senescence while did not lead to apoptosis. Compared with control group, cell viability was decreased, the apoptotic rate、levels of ROS and the mRNA of caspase-3, caspase-9 and Bcl-2 were increased in H/R and PC groups (<0.01). There were no differences in the above indexes between PC group and H/R group. Supplementation of NaHS increased cell viability and decreased apoptotic rate and oxidative stress. The effects of PC + NaHS on the above indexes were better than those of H/R+NaHS group.@*CONCLUSIONS@#Exogenous HS can restore the protective effect of PC on the aged H9C2 cells, and its mechanism is related to the inhibition of oxidative stress and apoptosis.
Subject(s)
Humans , Apoptosis , Cell Hypoxia , Cell Survival , Hydrogen Peroxide , Myocytes, Cardiac , Reactive Oxygen SpeciesABSTRACT
Gardner syndrome is a rare autosomal dominant disease. Its symptoms include multiple intestinal polyps, soft tissue tumors, dental disorders, osteoma, and congenital hypertrophy of the retinal pigment epithelium. Here, we present a patient with Gardner syndrome and chronic osteomyelitis of the jaw to highlight the serious damage that can be caused by Gardner syndrome.
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Objective To study the serum pharmacochemistry of Liujing Toutong Tablets (LTT). This study could be helpful to elucidate the bioactive constituents of LTT. Methods HPLC-Q/TOF-MS was used to analyze the ingredients in rats blood plasma with LTT (ig). Results By comparing the information on the total ion chromatogram, extraction chromatogram and the mass spectrogram of LTT, rat serum with drug and blank serum sample was used to confirm the constituents of LTT in vivo. As a result, a total of 46 compounds were detected in rat plasma, including 24 absorbed prototype constituents, such as puerarin, and 22 of the metabolites. Conclusion The compounds are absorbed into the blood might function as real bioactive constituents and this study can provide a scientific fundament for bioactive substances of LTT.
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Objective To establish the HPLC fingerprint of Liujing Toutong Tablets (LTT) and multi-wavelength method for determination of the contents of 3’-hydroxy puerarin, puerarin, daidzin, nuezhenide, and daidzein, so as to provide a reference for the quality control. Methods Using the method of HPLC, the fingerprint chromatography of high polarity and low polarity were established, with a mobile phase of acetonitrile-0.1% phosphoric acid and Diamonsil C18 column (250 mm × 4.6 mm, 5 μm). Furtherly, the Diamonsil C18 column was used with a mobile phase of acetonitrile-0.1% acetic acid gradient elution to determine the five contents. Results The fingerprint chromatography of high and low polarities with 11 batches of LTT were established, and the similarities of 11 batches were all over 0.90. The low polarity fingerprint chromatography included 16 mutual peaks, 12 of which belonged to herbs: No. 11, 12, 13 belonged to Angelicae Dahuricae Radix, No. 1, 2, 3, 7 belonged to Puerariae Lobatae Radix, No. 8 belonged to Ligustri Lucidi Fructus, No. 5, 6 belonged to Chuanxiong Rhizoma, No. 15 belonged to Ligustici Rhizoma et Radix and No. 4 belonged to Chuanxiong Rhizoma and Ligustici Rhizoma et Radix. Seven peaks including puerarin (No. 1), 3’-methoxy puerarin (No. 2), daidzin (No. 3), ferulic acid (No. 4), daidzein (No. 7), imperatorin (No. 11), and isoimperatorin (No. 13) were verified by standard compounds. The high polarity fingerprint chromatography included 10 mutual peaks, among them No. 2-9 belonged to Puerariae Lobatae Radix, No. 1, 10 belonged to Ligustri Lucidi Fructus. Five peaks including 3’-hydoxypuerarin (No. 2), puerarin (No. 3), 3’-methoxypuerarin (No. 4), daidzin (No. 7) and specnuezhenide (No. 10) were verified by standard compounds. The five active components were well separated and showed good lineaeity, such as 3’-hydroxy puerarin 71.60-716.00 ng (r = 0.999 1), puerarin 407.78-4 077.80 ng (r = 0.999 4), daidzin 90.72-907.20 ng (r = 0.999 9), nuezhenide 95.80-958.00 ng (r = 0.999 1), daidzein 21.98-219.80 ng (r = 0.999 9). The average recoveries and corresponding RSD values were 101.1% (1.38%), 97.8% (0.72%), 99.1% (0.75%), 97.8% (2.75%), and 98.7% (0.70%). The contents of 3’-hydoxypuerarin, puerarin, daidzin, specnuezhenide and daidzein in the 10 batches of sample were 3.08-3.84 mg/mL, 17.71-21.24 mg/mL, 3.51-4.71 mg/mL, 1.40-5.69 mg/mL, and 0.66-0.86 mg/mL, respectively. Conclusion The HPLC fingerprint combined with multi index determination method can reflect the intrinsic quality of LTT, which is stability, accurate, and reproducible, providing a scientific reference for quality control.
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Objective: To establish the HPLC fingerprint and to determine multiple components of leaves of Diospyros kaki, in order to provide a scientific basis for quality control for the D. kaki leaves and related products. Methods: The chromatographic separate was achieved on a Diamonsil C18 (250 mm × 4.6 mm, 5 μm) column with acetonitrile and 0.2% formic acid in water as the mobile phase for gradient elution; Volume flow rate was 1.0 mL/min; Column temperature was 35 oC; Detection wavelength was 360 nm; And injection was 10 μL; On 22 batches of persimmon leaf medicinal materials of fingerprint study, was evaluated with a chromatographic fingerprint similarity evaluation system (version 2004A), the 22 batches of persimmon leaf are divided into two categories by system cluster analysis and PCA analysis. And the determination method of hyperoside, isoquercitrin, quercetin, and kaempferol was also investigated at the same time. Results: The fingerprint of D. kaki leaves was established. There were 12 common peaks in the fingerprint of D. kaki leaves. Hyperoside, isoquercitrin, quercetin and kaempferol were baseline seperated with good linearity relationships between concentration and peak areas over the linear ranges, within 0.044-0.880, 0.078-1.560, 0.040-0.800 and 0.035-0.700 μg(r > 0.999 8). The average recoverys of the investigated compounds were 97.59%, 98.97%, 100.55%, 99.96%. Conclusion: HPLC fingerprint binding index component determination can provide a reference for the quality of D. kaki leaves and related products control.
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<p><b>OBJECTIVE</b>To investigate the association between rs751141 gene polymorphisms in EPHX2 gene and essential hypertension in Kazak and Han in Xinjiang.</p><p><b>METHODS</b>A total of 267 essential hypertensive patients in Kazaks, 368 essential hypertensive patients in Hans, 284 normotensive controls in Kazaks and 348 normotensive controls in Hans were enrolled in this study. TaqMan assay was used to detect the rs751141 G/A gene polymorphisms of EPHX2 gene.</p><p><b>RESULTS</b>The rs751141 G/A genotype frequencies for GA + AA genotypes was 40.2 percent in essential hypertensive subjects and 52.0 percent in control subjects in Hans, respectively. The genotype frequencies were significant difference between the two groups in Hans in Xinjiang (P < 0.01). The rs751141G/A gene polymorphism had no significant difference between essential hypertensive patients and normotensive controls in Kazaks in Xinjiang (P > 0.05).</p><p><b>CONCLUSION</b>The essential hypertension in Kazaks in Xinjiang is not associated with rs751141G/A gene polymorphism of EPHX2 gene, but the essential hypertension in Hans in Xinjiang is associated with rs751141G/A allele gene polymorphism of EPHX2 gene. A type of rs751141 allele gene polymorphism may be the independent protective factor of essential hypertension in Hans in Xinjiang.</p>
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Adult , Humans , Middle Aged , Alleles , Asian People , Genetics , China , Epidemiology , Epoxide Hydrolases , Genetics , Gene Frequency , Genotype , Hypertension , Epidemiology , Genetics , Polymorphism, Single NucleotideABSTRACT
Objective To find the effective therapeutic arrangement through investigating the clinical characteristics of chronic cardiac insufficiency with anaemia. Methods A total of 46 cases of anemia from 315patients who had been admitted to department of cardiology, the First Affiliated Hospital, Harbin Medical University for chronic cardiac insufficiency with anaemia were selected. They were divided into two groups. There were 22 patients in the first group who only accepted treatment to improve cardiac function (normal cardiac, diuretic,vasodilator therapy, etc.), and 24 patients in the second group who accepted treatment to improve cardiac function while receiving anti-anemia therapy treatment, oral ferrous sulfate tablets(0.3 g/tablet), 1 tablet each time, 3 times a day and(or) 2 times per week subcutaneous erythropoietin 3000 U. The hemoglobin(Hb), red blood cell(RBC) ,hematocrit (HCT), left ventricular ejection fraction (LVEF), fractional shortening (FS), stroke volume (SV) , cardiac output(CO) and E peak and A peak ratio(E/A) were observed before and after treatment. By logistic regression, grade grade Ⅱ , Ⅲ , Ⅳ, the incidence of anaemia were 7.9% (10/126), 19.2% (23/120) and 24.6% (17/69),respectively. Grade Ⅱ compared with grades Ⅲ, Ⅳ, the difference was statistically significant (x2 = 4.08, 3.12, all (3.49 ± 0.17) × 1012/L, (0.36 ± 0.08)%, (48.9 ± 3.11)%, (15.6 ± 1.8)%, (38.9 ± 3.7)%, (4.4 ± 1.6)% and (130.7 ±5.75)g/L, (4.12 ± 0.25) × 1012/L, (0.43 ± 0.02)%, (58.5 ± 2.65)%, (18.0 ± 2.5)%, (49.1 ± 7.7)%, (5.1 ± 1.2)%in the first and second groups, respectively. The difference between the two groups was statistically significant(t =value of Hb, RBC, HCT, LVEF, FS,SV, CO were (102.7 ± 6.93)g/L, (3.41 ± 0.12) × 1012/L, (0.35 ± 0.07)%,(47.5 ± 2.86)%, (16.0 ± 2.4)%, (38.2 ± 7.9)%, (3.7 ± 1.4)%, respectively. Compared with those after treatment,the difference was statistically significant (t = 15.632, 13.325, 5.569, 17.182, 3.186, 2.999, 3.074, all P < 0.05);Ⅳ-level relative risk were 1.62, 3.14(P < 0.05 or < 0.01) . Conclusions Based on the standard treatment with treatment of anemia, cardiac contractile function can be improved.
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<p><b>OBJECTIVE</b>To explore the expression of III β-tubulin and MDR1 protein in patients with non-small cell lung cancer (NSCLC), and to clarify its clinical significance.</p><p><b>METHODS</b>Paraffin embedded tissues from 158 primary non-small cell lung cancers and para-cancerous lung tissues were investigated for the expression of III β-tubulin and MDR1 protein by immunohistochemistry, as well as in freshly-taken NSCLC tissues by Western blot. The relationship between the expression of III β-tubulin and MDR1 and the biological features of lung cancer was analyzed.</p><p><b>RESULTS</b>The positive rate of III β-tubulin and MDR1 protein expression in lung cancer tissues was 65.19% and 51.27%, respectively. Western blot analysis showed that the level of of III β-tubulin and MDR1 protein in NSCLC tissues was remarkably higher than that in normal tissues (P < 0.01). The expression of III β-tubulin in stage III-IV cases was significantly higher than that in stage I-II cases (P < 0.05), while the expression of MDR1 protein showed no significant difference (P > 0.05). The positive rate of III β-tubulin expression in well-moderate pathological grades was lower than that in poor ones. The positive rate of MDR1 expression in adenocarcinoma was higher than that in squamous cell carcinoma and large cell undifferentiated cancers (P < 0.01). The positive rate of expression of MDR1 protein and III β-tubulin was not correlated with sex, age, tumor size and lymph node metastasis (P > 0.05).</p><p><b>CONCLUSION</b>The expression of III β-tubulin and MDR1 may play an important role in the development and progression of human non-small cell lung cancer, and could be looked as an important index for judging the prognosis of lung cancer.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Adenocarcinoma , Metabolism , Pathology , Carcinoma, Large Cell , Metabolism , Pathology , Carcinoma, Non-Small-Cell Lung , Metabolism , Pathology , Carcinoma, Squamous Cell , Metabolism , Pathology , Lung Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Neoplasm Staging , Prognosis , Tubulin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>to investigate the effects of clotrimazole on the growth of oral squamous cell carcinoma (OSCC).</p><p><b>METHODS</b>OSCC-25 cells growing in log phase were treated with various doses of clotrimazole. The concentration of IC(50), cell cycle and cell cycle related protein were examined.</p><p><b>RESULTS</b>the concentration of clotrimazole for inhibiting OSCC was IC(50) 8.51 µmol/L. Clotrimazole induced cell cycle arrest in the G(0)-G(1) cell cycle phase, with a concomitant decrease of cells in the G(2)-M and S-phase. Furthermore, clotrimazole significantly decreased the levels of cyclin D, cyclin E and CDK-4.</p><p><b>CONCLUSIONS</b>clotrimazole inhibits the growth of OSCC.</p>
Subject(s)
Humans , Carcinoma, Squamous Cell , Pathology , Cell Cycle , Cell Cycle Checkpoints , Cell Division , Cell Line, Tumor , Clotrimazole , Pharmacology , Cyclin E , Mouth Neoplasms , Pathology , Oncogene ProteinsABSTRACT
Objective To find out pathologieal change and the expression of neuron specific enolage (NSE)in the hippocampus dentate gyrus granular cell layer of epilepsy patients,to investigate the relationship between the pathological change and the cause of the epilepsy.Methods The specimens of hippocampus were from 9 epilepsy patients and 20 normal persons and the pathological change were investigated under the staining of the hematoxylin and eosin staining.The expression of NSE in the hippocampns denmte gyrus granular cell layer of epilepsy patients was detected by immunohistochemistry(IHC)with specific antibody,and the rate of NSE positive nenrons was evaluated.Results The nuclear pyknosis was observed in all of hippocampus from the epilepsy patients and some neurons were swelling.The positive of NSE was showed to have yellow granule;the rate of NSE positive was 28.66%.The expression of NSE of the neurons in the hippocampus dentate gyrus granular cell layer was.significantly reduced in epilepsy patients(7.9±5.6)%.The normal neuron nuclear was big and round in the middle of the cell and the nucleolus could be seen easily.The expression of NSE of the neurons in the hippocampus dentate gyrus granular cell layer was(39.0±17.4)%.Compared with normal group,the number of neurons with yellow granule was reduced.The difference of the NSE expression rate between the two group was statistically significant(t=-5.13,P<0.01).Conclusions The result suggests that the pathological abnormality and the reduced expression of NSE on the hippocampus dentate gyrus granular cell layer neurons could be one of the main reasons of the occurring of epilepsy.
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Objective To observe the variation of enzymatic activity and areas and bulk of focus of heart injuries by using controllable catheter to ablate epicardial tmsue of rabbits and focus underneath atrioventrieular ring narcosis with 20% urethane(4 ml/kg)and divided into three groups.Each group included 7 rabbits.Anterior wallepieardium of left ventricle was ablated thirty seconds in each group(10,20 and 30 W)with self-made ablationspheroid microwave antenna,refilling with high pressure normal saline at same time.Then all of the rabbits were sacrificed respectively and their ventricular myocardium were taken out to undergo immunohistochemistry in order to display suceinate dehydrogenase(SDH).Also amplitude Wag measured in order to calculate areas of heart injuries.(8F)wag delivered to the pre-selected sites around atrioventricular ring of thirty-two healthy dogs,which had beenin intravenous narcosis with pentobarbital sodium(30 mg/kg).The dogs were divided into four groups(40,50,60 and 80 w) and two time points(60 and 120 s),by the combined method of X-ray and endocardial electrocardiograph,the microwave antenna could be confirmed to be located at the accurate position between anterior and posterior wall close to septum of left/right ventricle.After ventricular myocardium had been taken out,amplitude were measuredin order to calculate bulk of heart injuries by 1/6×3.14 x long×wide×deep.In addition.the histological changesand transmural injury were examined by optic microscope.Results In each group,the centre of injuries wagenzyme deficiency locus.The diameter and areag of heart injuries enlarged significantly(3.99.±0.41),(5.20±0.25),(6.31±0.37)mm and(12.53±2.56),(21.19±3.14),(30.96±3.76)mm2 with the increased microwave power level(10、20、30 W).Group comparison had statisficM significance(F=76.8,58.5;P<0.01 or <0.05).A total of 116points were ablated.The myocardial lesion showed ellipse in shape,and continuous symmetrical coagulationnecrosis under microscopic examination.There was a clear demarcated line around tlle myocardial tissue and fewparietal thmmbus.There were 16 transmura]injuries and five-with lung damage.The bulk of lesion aroundatrioventrieular ring hag been significantly enlarged(46.7±2.5),(51.1±2.7),(133.2±3.4),(141.8±3.9),(248.5±6.2),(260.3±6.5),(313.7±9.5),(327.4±10.5)with the increased microwave power level(40,50,60and 80 W)and/or distance of microwave ablation(60 and 120 s).Groups comparison had statistical significance(F=31.16,27.85;all P<0.01).In each time point,the lesion bulks had conspicuous distinction of statistics.In the same microwave power,the time wag longer,the bulk was larger(P<0.01).Conclusions The more the microwave power level and time,the severe the heart injuries is.It is possible to use the microwave energy to ablate the deep focus under endocardium around atrioventricular ring.
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<p><b>OBJECTIVE</b>To investigate whether chemically synthesized double-stranded RNA (dsRNA) targeting epidermal growth factor receptor (EGFR) can induce gene silencing in non-small cell lung cancer (NSCLC) cells in vivo.</p><p><b>METHODS</b>The NSCLC cell line SPC-A1 was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000. SPC-A1 cells (1 x 10(7)/ml) in 200 microl were injected s.c. into the left flank area of the athymic nude mice to establish a tumor-bearing nude mouse model. To calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor. Immunohistochemistry and Western blot were used to monitor the reduction of EGFR protein production. Real-time RT-PCR was used to detect the silencing of the EGFR mRNA level.</p><p><b>RESULTS</b>The EGFR sequence specific dsRNA (dsRNA-EGFR) significantly inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.0%. The dsRNA-EGFR sequence specifically silenced EGFR with 53.6% of down-regulation of EGFR protein production and 32.3% of silencing of EGFR mRNA level.</p><p><b>CONCLUSION</b>dsRNA-EGFR show a blockbuster effect in down-regulation of EGFR mRNA level and protein production, and inhibition of tumor growth in vivo.</p>
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Animals , Humans , Male , Mice , Carcinoma, Non-Small-Cell Lung , Metabolism , Pathology , Cell Line, Tumor , Down-Regulation , Gene Expression Regulation, Neoplastic , Lung Neoplasms , Metabolism , Pathology , Mice, Nude , RNA Interference , RNA, Double-Stranded , Genetics , RNA, Messenger , Metabolism , Random Allocation , ErbB Receptors , Genetics , Transfection , Tumor BurdenABSTRACT
When albino rats were fed with high cholesterol diet or injected With a large dose of carbon tetrachloride or chloroform, severe fatty liver developed but without any morphological changes in the pancreas. However, when fatty liver was produced with a protein free diet, the atrophic change in the acinar cells was observed prior to fatty infiltration in the liver. No change was found in the islets of Langerhans.The presence of lipocaic factor as an internal secretion of the pancreas cannot be verified by these studies or by our previous works.