Shikonin induces hepatocellular carcinoma cell apoptosis by suppressing PKM2/PHD3/HIF-1α signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism of shikonin-induced death of human hepatocellular carcinoma SMMC-7721 cells.@*METHODS@#Cultured SMMC-7721 cells and normal hepatocytes (L-02 cells) were treated with 4, 8, or 16 μmol/L shikonin, and the changes in cell viability was assessed using MTT assay. The levels of ATP and lactic acid in the cell cultures were detected using commercial kits. Co-immunoprecipitation and immunofluorescence staining were used to determine the relationship among pyruvate kinase M2 (PKM2), prolyl hydroxylase 3 (PHD3), and hypoxia-inducible factor-1α (HIF-1α). The expressions of PHD3, PKM2, HIF-1α, Bax, cleaved caspase-3, and Bcl-2 in SMMC-7721 cells were detected with Western blotting, and cell apoptosis was analyzed with annexin V-FITC/PI staining. The effects of RNA interference of PKM2 on PHD3 and HIF-1α expressions in SMMC-7721 cells were detected using Western blotting.@*RESULTS@#The IC50 of shikonin against SMMC-7721 and L-02 cells was 8.041 μmol/L and 31.75 μmol/L, respectively. Treatment with shikonin significantly inhibited the protein expressions of PKM2, HIF-1α and PHD3 and nuclear translocation of PKM2 and HIF-1α in SMMC-7721 cells. Coimmunoprecipitation and immunofluorescence staining confirmed that shikonin inhibited the formation of PKM2/PHD3/HIF-1α complex and significantly reduced the contents of lactic acid and ATP in SMMC-7721 cells (P < 0.05). The expressions of PHD3 and HIF-1α decreased significantly after PKM2 knockdown (P < 0.05). Shikonin treatment significantly increased the apoptosis rate, enhanced the expressions of Bax and cleaved caspase-3, and decreased Bcl-2 expression in SMMC-7721 cells (P < 0.05).@*CONCLUSIONS@#Shikonin induces apoptosis of SMMC-7721 cells possibly by inhibiting aerobic glycolysis through the PKM2/PHD3/HIF-1α signaling pathway to cause energy supply dysfunction in the cells.

Risk factors for early in-hospital death in patients with acute Stanford type A aortic dissection / 中国胸心血管外科临床杂志

@#Objective     To investigate the risk factors for early in-hospital death in patients with acute Stanford type A aortic dissection and emergency surgical treatment. Methods    We retrospectively analyzed the clinical data of 189 patients with acute Stanford type A aortic dissection who underwent surgery in the First Affiliated Hospital of Xinjiang Medical University between January 2017 and January 2020. There were 160 males and 29 females with an average age of 46.35±9.17 years. All patients underwent surgical treatment within 24 hours. The patients were divided into a survival group (n=160) and a death group (n=29) according to their outcome (survival or death) during hospitalization in our hospital. Perioperative clinical data were analyzed and compared between the two groups. Results     The overall in-hospital mortality was 15.34% (29/189). There was a statistical difference between the two groups in white blood cell count, blood glucose, aspartate aminotransferase (AST), bilirubin, creatinine, operative method, operation time, aortic occlusion time, or cardiopulmonary bypass time (P<0.05). Multivariate regression identified white blood cell count [OR=1.142, 95%CI (1.008, 1.293)], bilirubin [OR=0.906, 95%CI (0.833, 0.985)], creatinine [OR=1.009, 95%CI (1.000, 1.017)], cardiopulmonary bypass time [OR=1.013, 95%CI (1.003, 1.024)] as postoperative risk factors for early in-hospital death in the patients undergoing acute Stanford type A aortic dissection surgery (P<0.05). Conclusion     Our study  demonstrated that white blood cell, bilirubin, creatinine and cardiopulmonary bypass time are independent risk factors for in-hospital death after acute Stanford type A aortic dissection surgery.

Comparison of diagnostic values between myocardial perfusion imaging and dual source CT coronary angiography in coronary artery stenosis in patients with diabetes complicated with coronary heart disease / 吉林大学学报(医学版)

Objective:To evaluate and compare the diagnostic values between myocardial perfusion imaging (MPI)and dual source CT coronary angiography(DS-CTCA)in coronary artery stenosis in the diabetic patients with coronary heart disease(CHD),and to elucidate the clinical application values of MPI combined with DS-CTA in the diabetic patients with CHD.Methods:A total of 52 diabetic patients with CHD underwent the examinations of MPI and DS-CTCA were selected.The degrees of coronary artery stenosis of branches of coronary artery were compared between MPI examination and DS-CTCA examination,including left main coronary artery(LM),left anterior descending coronary artery(LAD),left circumflex branch(LCX)and right coronary artery(RCA).Based on the coronary angiography(CAG)regarded as the golden standard,the specificity,sensitivity and accuracy of each imaging examination method were compared;the specificities,sensitivities and accuracies of parallel diagnosis and serial diagnosis of two kinds of methods were also compared.Results:The stenosis degrees of LM and LAD were no different between MPI examination and DS-CTCA examination(P>0.05),but the stenosis degrees of LCX and RCA were different between two imaging examination methods(P<0.05).Compared with MPI examination, the sensitivity of DS-CTCA examination in the diabetic patients with CHD was decreased(71.0%vs 90.1%),and the difference was significant(P=0.035);the specificity of DS-CTCA examination in the diabetic patients with CHD was increased(85.7%vs 58.4%),and the difference was significant(P=0.027);but the accuracies of two examinations in the diabetic patients with CHD were no different(76.4%vs 78.4%)(P=0.062).Compared with parallel diagnosis,the specificity and accuracy of serial diagnosis were increased(93.5% vs 33.8%,P=0.001;94.7%vs 71.2%,P = 0.030);but the sensitivities had no difference(95.4% vs 93.1%,P = 0.074). Conclusion:The diagnostic accuracy of evaluating the degree of coronary artery stenosis in the diabetic patients with CHD is not different between DS-CTCA examination and MPI examination. And the serial diagnosis of two examination methods can improve the diagnostic accuracy.

Enzymatic biosynthesis of novel neobavaisoflavone glucosides via Bacillus UDP-glycosyltransferase / 中国天然药物

The present study was designed to perform structural modifications of of neobavaisoflavone (NBIF), using an in vitro enzymatic glycosylation reaction, in order to improve its water-solubility. Two novel glucosides of NBIF were obtained from an enzymatic glycosylation by UDP-glycosyltransferase. The glycosylated products were elucidated by LC-MS, HR-ESI-MS, and NMR analysis. The HPLC peaks were integrated and the concentrations in sample solutions were calculated. The MTT assay was used to detect the cytotoxic activity of compounds in cancer cell lines. Based on the spectroscopic analyses, the two novel glucosides were identified as neobavaisoflavone-4'-O-β-D-glucopyranoside (1) and neobavaisoflavone-4', 7-di-O-β-D-glucopyranoside (2). Additionally, the water-solubilities of compounds 1 and 2 were approximately 175.1- and 4 031.9-fold higher than that of the substrate, respectively. Among the test compounds, only NBIF exhibited weak cytotoxicity against four human cancer cell lines, with IC values ranging from 63.47 to 72.81 µmol·L. These results suggest that in vitro enzymatic glycosylation is a powerful approach to structural modification, improving water-solubility.

Biosynthesis of a new psoralidin glucoside by enzymatic glycosylation / 南方医科大学学报

<p><b>OBJECTIVE</b>To modify the structure of psoralidin using in vitro enzymatic glycosylation to improve its water solubility and stability.</p><p><b>METHODS</b>A new psoralidin glucoside (1) was obtained by enzymatic glycosylation using a UDP- glycosyltransferase. The chemical structure of compound 1 was elucidated by HR-ESI-MS and nuclear magnetic resonance (NMR) analysis. The high-performance liquid chromatography (HPLC) peaks were integrated and sample solution concentrations were calculated. MTT assay was used to detect the cytotoxicity of the compounds against 3 cancer cell lines in vitro. Results Based on the spectroscopic data, the new psoralidin glucoside was identified as psoralidin-6',7-di-O-β-D- glucopyranoside (1), whose water solubility was 32.6-fold higher than that of the substrate. Analyses of pH and temperature stability demonstrated that compound 1 was more stable than psoralidin at pH 8.8 and at high temperatures. Only psoralidin exhibited a moderate cytotoxicity against 3 human cancer cell lines. Conclusion In vitro enzymatic glycosylation is a powerful approach for structural modification and improving water solubility and stability of compounds.</p>

Parathyroid in situ exposure and protection in differentiated thyroid carcinoma surgery / 中华内分泌外科杂志

Objective To study the application value of parathyrod in situ exposure and protection in differentiated thyroid cancer thyroid carcinoma (DTC) surgery. Methods 500 cases of DTC admitted from Jan. 2012 to Dec. 2014 were selected for the research. According to whether the parathyroid was anatomically exposed during the surgery, the patients were divided into experimental group (exposed group) and the control group (un-exposed group). Then, depending on tumor extent and risk factors, the experimental group were divided into group E1 (lateral affected glandular lobe, isthmus lobe resection and unilateral VI lymph node dissection )and E2 (total or subtotal thyroidectomy and bilateral VI lymph node dissection), while the control group were divided in-to group C1 (surgical treatment same as E1)and group C2 (surgical treatment same as E2). Patients’ postopera-tive nerve and muscle symptoms were observed, such as clinical manifestations of face, lips, hand and foot numb-ness or convulsions. Both preoperatively and postoperatively, patients’ parathyroid hormone (PTH) and serum calcium level indicators were detected before surgery and at the 1st, 3rd, 5th, 7th, 14th and 28th day after surgery. Results The lymph node metastasis rate and number between experimental group and the control group had statistical insignificance (χ2=1.02, 0.79, P=0.14, 0.96). The recurrence rate of the 2 groups had no significant difference during the follow-up (χ2=0.23, P=0.65). Group E1 was significantly lower than Group C1 in postopera-tive parathyroid function decline rate, incidence of hypocalcemia and parathyroid mistakenly cut rate. Group E2 was also obviously lower than Group C2 in all these aspects. The difference had statistical significance (P<0.05). Conclusion Parathyrod in situ exposure and protection assisted by direct vision of the magnifying glass can re-duce the mistakenly cut rate of parathyroid and the temporary hypoparathyroidism, which can effectively avoid the adverse effects caused by total thyroidectomy.

Transglycosylation of neolignans by enzymatic synthesis and evaluation of their antitumor activity / 南方医科大学学报

<p><b>OBJECTIVE</b>To improve the water solubility and biological activity of neoligans (magnolol and honokiol) and test the antitumor activity of the modified compounds.</p><p><b>METHODS</b>The glycosylated products of magnolol and honokiol were obtained by enzymatic synthesis using a UDP-glycosyltransferase (YjiC) from Bacillus. The products were characterized by high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and nuclear magnetic resonance (NMR) analysis. MTT assay was used to detect the growth inhibition of 4 human cancer cell lines induced by the compounds.</p><p><b>RESULTS</b>We obtained two glucosides of neolignans (magnolol and honokiol) for the first time by enzymatic synthesis using a UDP-glycosyltransferase. Based on the spectroscopic data, the glucosides were identified as magnolol-2- O-β-D-glucopyranoside (1) and honokiol-4'-O-β-D-glucopyranoside (2). Compounds 1-4 exhibited moderate anti-proliferative activities against the 4 human cancer cell lines, with IC50 values ranging from 9.41 to 111.21 µmol/L.</p><p><b>CONCLUSION</b>The glycoslated products show enhanced water solubility and drug sensitivity against SMMC7721 cells, suggesting their value as potential therapeutic drugs.</p>

Effect of anacardic acid, a Hsp90 inhibitor, on proliferation, invasion and migration of breast cancer MDA-MB-231 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of the Hsp90 inhibitor anacardic acid on cell proliferation, invasion and migration of breast cancer MDA-MB-231 cells.</p><p><b>METHODS</b>The inhibitory effect of anacardic acid on Hsp90 was assessed with in vitro ATPase inhibition assay and ATP-sepharose binding assay. MTT assay was used to detect the growth inhibition induced by anacardic acid in MDA-MB-231 cells. Transwell assays were used to evaluate MDA-MB-231 cell invasion and migration. Western blotting was performed to assess the effect of anacardic acid in triggering the degradation of MMP-9, TIMP-1, Hsp90, and Hsp70.</p><p><b>RESULTS</b>Anacardic acid exhibited a modest activity of ATPase inhibition with an IC50 value of 82.5 µmol/L. Anacardic acid significantly suppressed the proliferation of MDA-MB-231 cells in a dose-dependent manner (IC50 value of 29.3 µmol/L). Treatment with 12.5, 25, and 50 µmol/L anacardic acid for 36 h caused inhibition of cell invasion by 23.6%, 56.6%, and 67.0% in MDA-MB-231 cells, respectively (P<0.05), and anacardic acid treatment for 24 h inhibited the cell migration by 30.0%, 45.5%, and 77.5%, respectively (P<0.05). Anacardic acid dose-dependently induced MMP-9 degradation, but did not obviously affect Hsp90 or Hsp70 expressions.</p><p><b>CONCLUSION</b>Anacardic acid can significantly inhibit the proliferation, invasion, and migration of MDA-MB-231 cells, the mechanism of which may involve the inhibition of Hsp90 ATPse activity and down-regulation of MMP-9 expression.</p>

Surgery for sinus of Valsalva aneurysm: 33-year of a single center experience / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sinus of Valsalva aneurysm (SVA) is a rare anomaly and few large or long-term series are well established. This study was designed to review 33-year surgical experience of SVA in one center.</p><p><b>METHODS</b>From August 1980 to December 2013, patients with SVA underwent surgical repair were retrospectively studied.</p><p><b>RESULTS</b>A total of 160 patients were identified with mean age of (30±12) years and 112 (70%) of them were males. The right coronary sinus origin of SVA was found in 108 patients (67.5%), the non-coronary sinus in 51 patients (31.9%), and the left coronary sinus in one patient (0.6%). The rupture of SVA into the right ventricle was identified in 89 (55.6%) cases, the right atrium in 61 (38.1%), the left ventricle in 2 (1.3%) and no rupture in 8 (5.0%). Ventricular septal defect (VSD) and aortic regurgitation (AR) were found in 59 (37%) and 45 (28%) patients respectively. An approach via the involved chamber was used in 86 patients (54%), aortotomy in 8 (5%), and a combined approach in 66 (41%). Either direct suture (56, 35%) or patch (104, 65%) closure were used to repair the SVA. The VSD was closed with a patch (44/59, 75%) or direct suture (15/59, 25%). aortic valve replacement (AVR) was performed in 23/45 (51%) and aortic valvuloplasty (AVP) in 9/45 (20%) patients combined with AR. There were 3 hospital deaths (1.9%) and 2 late deaths and 84% of the patients were followed up for (17.6±4.2) years. New York Heart Association functional class improved significantly after surgery (P < 0.01). Actuarial survival was 94% at 10 years, and 88% at 20 years.</p><p><b>CONCLUSIONS</b>Surgical treatment of SVA is safe and effective, ruptured SVA or unruptured SVA with VSD and/or AR should be repaired surgically as early as possible. However, late progressive AR is still a risk during long-term follow-up, and early aggressive measures are recommended. These include more use of a combined approach to achieve optimal evaluation of lesions, more patch repairs to reduce the chance of recurrence, and more AVR in patients with moderate to severe AR.</p>

Construction and screening of specific short hairpin RNA vector targeting heparanase gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct short hairpin RNA (shRNA) expression vectors of RNA for specific silencing of heparanase (HPA) gene.</p><p><b>METHODS</b>The genomic sequence of HPA gene was retrieved from GenBank and the cDNA encoding shRNA for HPA gene silencing was designed. Five specific interference sequences and a random negative control sequence were inserted into the vector pGPU6/GFP/Neo. After verification by restriction enzyme digestion and sequence analysis, the recombinant vectors were transfected into MDA-MB-231 cells via lipofectamine. Fluorescent quantitative PCR and Western blotting were employed to detect the expression of HPA gene expressions in the transfected cells at the mRNA and protein levels, respectively.</p><p><b>RESULTS</b>Both restriction analysis and sequencing confirmed correct construction of the shRNA vectors. Transfected with the specific siRNA vectors HPSE-1 and HPSE-5 resulted in significantly decreased expression level of HPA protein in MDA-MB-231 cells, while negative control vector produced no significant changes in HPA expressions.</p><p><b>CONCLUSION</b>We have obtained two shRNA vectors which can significantly down-regulate HPA expressions in MDA-MB-231 cells, which facilitates further investigation of the role HPA may play in the invasiveness and metastasis of human breast cancer.</p>

Differential expression of collagen and matrix metalloproteinases between left and right atria in patients with chronic atrial fibrillation / 生理学报

The present study aimed to investigate the mRNA expression levels of collagen, matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs) in the left and right atria in patients with chronic atrial fibrillation (CAF). Forty five patients with valvular heart disease were sampled in this study, including 18 patients with sinus rhythm (SR), 27 patients with CAF. Clinical data of these patients were collected, and the left and right atrial appendages were obtained from these patients during heart valvular replacement surgery. The mRNA levels of collagen type I, collagen type III, MMP1, MMP9, TMP1 of the atria were then measured by semi-quantitative RT-PCR. The results obtained were as follows. Compared to those in SR group, in atria of CAF group, the mRNA levels of collagen type I, MMP1 and MMP9 increased (P<0.05), while the mRNA level of TMP1 decreased (P<0.01). There were no significant differences in the mRNA levels of collagen type I, collagen type III, MMP1, MMP9 and TMP1 between the left and right atria of SR group (P>0.05). In CAF group, the mRNA level of MMP1 in the right atrium was higher than that in the left atrium (P<0.05), however, the mRNA level of MMP9 in the left atrium was higher than that in the right atrium (P<0.01). In both the left and right atria, the mRNA of collagen type I was positively correlated with the corresponding atrial diameter; the mRNA of MMP1 and MMP9 was positively correlated with the mRNA of collagen type I, and was negatively correlated with the mRNA of TMP1. These results suggest that the increased level of collagen type I associated with selective upregulation of MMP1, 9 and downregulation of TMP1, 9 in the atrium might be the molecular basis of atrial interstitial fibrosis in patients with CAF. Moreover, during CAF development, there is difference in the expression of MMPs between the left and right atria.

Research on Isolating Bacteria by Using Magnetophoresis / 中国生物工程杂志

There are similarities between magnetotactic bacteria and Acidithiobacillus ferrooxidans (A. ferrooxidans) which isolated from Acid mine drainage(AMD). The weak magnetotaxis of some bioleaching bacteria isolated were found by microscope. A magnetophoresis apparatus was designed based on these weak magnetotaxis and be used to analysis the movement of these strains. The physiological properties of the anear magnetic field strain and removed magnetic field strain which isolated successfully by magnetophoresis apparatus have large difference. The nanometer magnetic particles was extract from the Acidithiobacillus ferrooxidans which purified by spread plate method from AMFS and its main elements are Fe and O by energy spectrum analysis. The results show that A. ferrooxidans have weak magnetotaxis and can be isolated by magnetophoresis. With the development of this new isolating method, the research of magnetotactic bacteria and bioleaching will get more benefit from it.

Pharmacokinetics and bioequivalence of piracetam tablets in healthy volunteers / 中国临床药理学与治疗学

AIM: To study the bioequivalence of piracetam tablets in Chinese healthy volunteers. METH-ODS: Twenty volunteers were randomly divided into two groups (test and reference), with double cross-over design and single-dose oral administration. The concentration of piracetam in serum was determined by HPLC. The main pharmacokinetic parameters were calculated and the bioequivalence was evaluated with DAS2.0 practical pharmacokinetics program. RESULTS: The pharmacokinetic parameters of piracetam tablets were as follows: t1/2 were 5.50±1.48 and 4.29±1.00 h, Cmax were 21.47± 6.27 and 20.96±5.10 mg·L-1, Tmax were 0.70±0.46 and 0.66± 0.36 h, AUC0-24h were 93.44± 16.61 and 96.67± 18.50 mg·h·L- 1. The relative bioavailability of the test preparation was 99.8%± 22.7%. CONCLU-SION: The test and reference preparations were bioequivalent and may be prescribed interchangeably.

Mini-thoracotomy for Open Heart Surgeries:Report of 810 Cases / 中国微创外科杂志

Objective To evaluate the value of mini-thoracotomy for open heart surgery.Methods From December 1995 to January 2008,810 patients including 660 cases of congenital heart diseases,129 cases of valvular heart diseases and 21 cases of heart myxoma,underwent open heart surgery by mini-thoracotomy through cardiopulmonary bypass in our hospital.Among the cases,superior median sternotomy was performed on 36 patients,inferior median sternotomy was made on 59,right parasternal mini-thoracotomy was carried out in 3,right anterolateral thoracotomy was done on 658,and right axillary mini-thoracotomy was used in 54.A total of 382 patients received beating-heart surgery.Results The postoperative mortality in our patients was 1.5%(12 cases).In this series,the mean postoperative mechanical ventilation time,drainage volume,and hospital stay were(6.7?4.2) hours,(210?165) ml,and(7.4?4.9) days respectively.421 of the patients received no blood transfusion.None of the patients developed sternal dehiscence or mediastinal infection.Follow-up was available in 690(85%) of the patients up to a mean of(48.2?25.3) months,none of them died during the period.The cardiac function of the patients was significantly improved after the operation(preparation vs postoperation: 310 cases vs 478 cases for grade Ⅰ,438 vs 212 for grades Ⅱ-Ⅲ,and 62 vs 0 for grade Ⅳ,Z=-13.21,P=0.000).The mean cardiothoracic ratio was decreased significantly after the operation(0.51?0.11 vs 0.53?0.08,t=4.065,P=0.000),while the left ventricular ejection fraction was increased markedly(0.63?0.11 vs 0.57?0.11,t=-10.529,P=0.000).Conclusions The mini-thoracotomy is superior in cosmetic results and the postoperative morbidity rates of sternal malformation and infection.Proper selection of patients,good exposure of the surgical field,and skillful surgical procedures are crucial for the outcomes of the operation.

Clinical characteristics and surgical treatment of cardiac and pericardial echinococcosis / 中华胸心血管外科杂志

Objective: To review clinical characteristics of cardiac and pericardial hydatid cysts and assess the results of surgical treatment. Methods: From February 1978 to July 1999, 13 patients were diagnosed as heart and/or pericardium echinococcosis. There were nine male and four female. Their age ranged from 8 to 36 years ［mean (23.0±8.5) years］. 10 patients were symptomatic and 3 were asymptomatic. 3 patients had involvement of liver or lung. Pathologic cardiac murmur or cardiac enlargements were found in 10 patients. All patients were examined with chest roentgenograms, electrocardiogram and echocardiography. The serologic test was positive in 11 cases and negative in 3 cases. One right atrial cyst was removed under extracorporal circulation. 4 single pericardial cyst and multi-pericardial cyst were resected, 6 myocardial cysts and one complicated myocard ial and pericardial cysts were resected through puncture-aspiration endocystect omy. Results: All patients were followed up from six months to six years ［mean (3.5±1.5) years］. One late death occurred ten-month after operation, and the cause of death was heart failure. 3 patients had recurrence and were reoperated. Conclusion: The clinical characters may vary according to the number, size and location of cysts. Echocardiography is important for surgical planning. The results of endocystectomy of cardiac and pericardial echinococcosis were excellent.