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@#Objective To provide experimental data and theoretical support for further studying the maturity of cardiac patches in other in vitro experiments and the safety in other in vivo animal experiments, through standard chemically defined and small molecule-based induction protocol (CDM3) for promoting the differentiation of human induced pluripotent stem cells (hiPSCs) into myocardium, and preliminarily preparing cardiac patches. Methods After resuscitation, culture and identification of hiPSCs, they were inoculated on the matrigel-coated polycaprolactone (PCL). After 24 hours, the cell growth was observed by DAPI fluorescence under a fluorescence microscope, and the stemness of hiPSCs was identified by OCT4 fluorescence. After fixation, electron microscope scanning was performed to observe the cell morphology on the surface of the patch. On the 1st, 3rd, 5th, and 7th days of culture, the cell viability was determined by CCK-8 method, and the growth curve was drawn to observe the cell growth and proliferation. After co-cultured with matrigel-coated PCL for 24 hours, hiPSCs were divided into a control group and a CDM3 group, and continued to culture for 6 days. On the 8th day, the cell growth was observed by DAPI fluorescence under a fluorescence microscope, and hiPSCs stemness was identified by OCT4 fluorescence, and cTnT and α-actin for cardiomyocyte marker identification. Results Immunofluorescence of hiPSCs co-cultured with matrigel-coated PCL for 24 hours showed that OCT4 emitted green fluorescence, and hiPSCs remained stemness on matrigel-coated PCL scaffolds. DAPI emitted blue fluorescence: cells grew clonally with uniform cell morphology. Scanning electron microscope showed that hiPSCs adhered and grew on matrigel-coated PCL, the cell outline was clearly visible, and the morphology was normal. The cell viability assay by CCK-8 method showed that hiPSCs proliferated and grew on PCL scaffolds coated with matrigel. After 6 days of culture in the control group and the CDM3 group, immunofluorescence showed that the hiPSCs in the control group highly expressed the stem cell stemness marker OCT4, but did not express the cardiac markers cTnT and α-actin. The CDM3 group obviously expressed the cardiac markers cTnT and α-actin, but did not express the stem cell stemness marker OCT4. Conclusion hiPSCs can proliferate and grow on matrigel-coated PCL. Under the influence of CDM3, hiPSCs can be differentiated into cardiomyocyte-like cells, and the preliminary preparation of cardiac patch can provide a better treatment method for further clinical treatment of cardiac infarction.
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Cirrhotic portal hypertension refers to a series of syndroms characterized by structural abnormality and dysfunction of hepatic sinusoid caused by chronic liver injury and obstructing portal-systemic blood flow, resulting in gradually increased portal venous system pressure as clinical manifestations. Increased intrahepatic resistance and portal venous system blood flow are main causes for cirrhotic portal hypertension. The structural abnormality and dysfunction of hepatic sinusoid cause not only increased intrahepatic resistance, but also substance exchange barriers between hepatic sinusoidal blood and hepatocytes, resulting in splanchnic artery dilation and increased blood flow and pressure of portal venous system. Dysfunction of splanchnic hemodynamic is an important factor for hyperdynamic circulation in cirrhotic portal hypertension. As the disease progresses, cirrhotic portal hypertension can continuously promote the activation of hyperdynamic circulation, which in turn can accelerate the development of cirrhotic portal hyperten-sion. This vicious circle is the main reason for the irreversible and untreatable end-stage liver disease. The authors review the pathophysiological mechanisms of cirrhotic portal hypertension, splanchnic hemodynamic dysfunction and hyperdynamic circulation.
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Objective:To compare the efficacy and safety of KeyPort access and traditional transanal endoscopic microsurgery(TEM) in the treatment of rectal tumors.Methods:In this study, 36 cases of rectal tumors were treated by KeyPort TEM access and 52 cases by traditional TEM. Tumor type, size, distance from anal edge, operation time, intraoperative blood loss, postoperative hospitalization time, specimen quality and complications were compared between the two groups.Results:There were no significant differences in tumor type, size, operation time, intraoperative blood loss, postoperative hospitalization time, complications, recurrence and metastasis rate between the two groups. The distance between lower cutting edge to the anus in KeyPort access group was significantly greater than that of traditional TEM group[(6.7±1.9) vs. (5.1±1.8) cm , t=3.901, P<0.001]. All the surgeries in the KeyPort access group were completed. While two cases of in traditional TEM group were coverted to other surgical approaches. All patients in the KeyPort group had normal anal function in the early postoperative period, while 2 patients in the traditional TEM group suffered anal function impairment. Conclusion:TEM by KeyPort access is safer and more effective then traditional TEM, as well as more generous indications.
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Prostate cancer (PCa) patients who progress to metastatic castration-resistant PCa (mCRPC) mostly have poor outcomes due to the lack of effective therapies. Our recent study established the orphan nuclear receptor ROR
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Objective To investigate the relationship between pelvic floor pain and pelvic floor injury in female patients with lower urinary tract disease.Methods Transvaginal and rectal pelvic floor acupressure was used to perform pelvic floor pain point examination in women with lower urinary tract disease.The number of tender points and the pain point of tenderness were recorded.R~ults The positive rate of pelvic pain points was 61.5％ (3 507 checkpoints,2 156 checkpoints had tenderness);there was a difference in the degree of pain caused by pain points in different regions (H =159.144,P ＜0.01).There was no difference in pain level between the P3 area,P4 and P5 areas and P2 (adjust P =1),and the pain points of the other areas were statistically different from the pain level (adjust P ＜ 0.01).Conclusions The density of tender points and degree of pain in pain points in female pelvic floor are related to the pelvic floor injury.It is of clinical significance to evaluate the degree of pelvic floor injury by taking the pelvic floor pain point.
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Objective To study the correlation between female urination abnormalities and pelvic floor injury.Methods A total of 193 adult women with urination abnormalities from January 2015 to January 2017 were selected from Foshan Maternal and Child Hospital of Guangdong Province,including 84 cases of lower urinary tract symptoms (LUTS),71 cases of overactive bladder (OAB) and 38 cases of stress urinary incontinence (SUI).All patients aged 38-82 years (mean 51 ± 3.74) were enrolled according to their clinical symptoms.100 adult female patients without obvious symptoms such as abnormal urination were selected as the control group.Pelvic floor pain points were examined and scored in both groups.Results In the 193 cases,the number of pain points in LUTS,OAB and SUI were 1 093,983 and 415,respectively,a total of 2 491.The total pain scores of pain points were 7 163,6 480 and 2 583,with a total of 16 226 points.There were 100 cases in the control group with 527 pain points,and the total pain points were 1 377 points.According to the pelvic floor injury formula,the injury scores of the inspection group and the control group were calculated to be 3.59 vs 0.24 respectively,with statistically significant difference (P ＜ 0.01).The pelvic floor injury scores of LUTS group,OAB group and SUI group were 3.67,4.18 and 2.46 respectively,with statistically significant difference (P ＜ 0.01).Conclusions Overall,female urination abnormalities are significantly associated with pelvic floor injury,which are consistent with clinical observation.
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BACKGROUND:In recent years, based on high-throughput molecular imaging, integration of genomics, proteomics and computer aided design and the application of correlative “technical chains” have achieved great achievements in the research of breast cancer, lung cancer, gastric cancer, colon cancer, ovarian cancer and melanin tumor. However, there are few researches on oral squamous cel carcinoma. OBJECTIVE:To detect the gene expression profile of the oral squamous cel carcinoma tissue and normal paracarcinoma tissue using DNA chip-based gene expression profile. METHODS:Two samples of oral squamous cel carcinoma tissue and normal paracarcinoma tissue of patients who received treatment at Stomatological Hospital of Guangdong Province of China in 2013 were included in this study. The gene expression profiles of oral squamous cel carcinoma and normal paracarcinoma tissue were determined by the Roche NimbleGen gene expression microarrays. RESULTS AND CONCLUSION: According to screening criteria of differential genes, 7 872 out of 32 448 detected genes were differentialy expressed genes of oral squamous cel carcinoma, which accounts for 24% of the total number of the screening genes. 3 800 genes were up-regulated, and 4 072 were down-regulated. The results confirm that through detection with the help of gene expression profile clip, 7 872 differentialy expressed genes were obtained through DNA chip-based gene expression profiles according to the screening criteria. Thus it can be concluded that the occurrence and development of the tumors are not a result of single or several genes. Previous experiments based on a single or several genes have great limitations. These findings also suggest that the occurrence of tumor is a result of mutual regulatory effects of many genes forming a network, moreover, the interactions of the network is quite complicated.
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Objective Investigate the reasonable surgical approach combined with frozen sections pathological features of papillary thyroid microcarcinoma.Methods Clinical data of 96 cases papillary thyroid microcarcinoma nearly 5 years of follow-up in our clinic referral was retrospectively analyzed.Metastasis and recurrence rate were compared between group of ipsilateral lobe plus isthmus resection (unilateral group) and group of ipsilateral lobe plus isthmus resection and contralateral lobe subtotal (bilateral group).Metastasis and recurrence rate were compared between group of central lymph node dissection (dissection group) and non-dissection group (non-dissection group),and the injury rate of the recurrent laryngeal nerve was also compared between the two groups.Results The diagnosis of cancer by intraoperative frozen pathology were 53 cases (55％).Whether in high or low risk populations,the metastasis and recurrence between unilateral and bilateral groups showed no significant difference (P ＞ 0.05).Whether in high or low risk populations,the metastasis and recurrence between dissection group and non-dissection groups showed no significant difference (P ＞0.05).The temporary injury rate of recurrent laryngeal nerve in dissection group were higher than thatin non-dissection group both in high-risk populations (P =0.040,P ＜ 0.05) and low risk populations(P =0.037,P ＜ 0.05).Conclusions Intraoperative frozen pathological diagnosis of papillary thyroid microcarcinoma is difficult.The reasonable surgical approach for the first time is ipsilateral lobe plus isthmus resection.Preventive cervical dissections operation should not be carried out if the exploratory of lymph node showed no metastasis.
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BACKGROUND:RACK1 is strongly associated with the occurrence and development of oral squamous cel carcinoma. However, the occurrence and development of tumor do not depend on a gene or protein, but a long-term complex process of a network structure of multiple genes and multiple molecules, multi-step, multi-stage joint action. Synergism between tumor genes promotes the formation and development of tumor cel s. Therefore, we cannot limit on a single gene or protein to discover the action mechanism of oral squamous cel carcinoma, but should pay attention on signaling network path related to differential protein or gene, investigate the alterations in related protein or gene expression in the whole signaling pathway, and analyze the action mechanism of the interaction of these molecules. OBJECTIVE:To screen differential genes related to oral squamous cel carcinoma, construct an interaction network through bioinformatics using STRING database, and provide clues for future tests. METHODS:In accordance with our previous classic proteomics results and microarray results of oral squamous cel carcinoma, genes with consistent expression and big differences were selected as differential genes. The differential genes were inputted into the database of STRING to find the possible relationship among the protein subunits and to construct network structure of their interaction. RESULTS AND CONCLUSION:The 19 differential proteins of oral squamous cel carcinoma construct a complicated net work, and the differential proteins interact through these networks. GNB2L1-encoded RACK1 is a node protein and interacts with other differential proteins via WD40 repeated protein (number COG2319) andβ-G protein subunit (number KOG0279). WD40 repeated protein (number COG2319) interacts with 5 differential proteins directly and constructs 10 interacting pathways.β-G protein subunit (number KOG0279) interacts with 8 differential proteins directly, which has 11 interacting pathways. We make a network structure picture based on the interaction of these 19 differential genes by the analysis of the STRING database. The results show that the two subunits of RACK1 protein have direct interaction with 8 differential proteins and have 18 interaction pathways on the picture. As a result, RACK1 is the core protein of the network, suggesting RACK1 is the key node protein in oral squamous cel carcinoma.
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Objective To evaluate the clinical efficacy of mesalazine sustained-release tablets in combination with azathioprine in the small intestine Crohn’s disease therapy.Methods 60 cases of small intestinal Crohn disease patients were randomly divided into two groups and the control group 30 cases.Observation group mesalazine sustained-release tablets 1.5 mg ~2.0 mg/( kg? d ) , 1 ~2 times/d, the other in combination with azathioprine 1 g/times, 2 times/d treatment;control group received azathioprine alone 1 g/times, 2 times/d, both groups were treated for 30 weeks after treatment were compared activity index score, clinical remission rate, mucosal healing rate, blood biochemical parameters before and after treatment, and record adverse reactions during treatment.ResuIts The two groups of patients after treatment than before treatment activity index score decreased significantly (P<0.05), but the observation group were more than the control group decreased significantly (P<0.05); observation group complete remission rate and mucosal healing rates better than the control group (P<0.05);the WBC count, ESR and CRP level were lower than before treatment (P<0.05);AIL two levels and TLB levels were significantly increased compared with that before treatment (P<0.01), but the observation group was significantly higher than that in the control group ( P<0.05 ); the incidence of adverse observation group were 26.67%, significantly higher than the 6.67%( P<0.05 ) .ConcIusion Mesalazine sustained-release tablets in combination with azathioprine therapy compared with small bowel Crohn’s disease treatment is more effective azathioprine alone, can significantly improve the therapeutic effect, but incidence of adverse reactions is higher.
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BACKGROUND:In recent years, endoplasmic reticulum stress-caused apoptosis plays a crucial role in ischemia impairment and has become the hotspot of studies addressing myocardial ischemia/reperfusion (I/R) injury. The lower limb ischemic preconditioning (LIP) has the obvious protective effect on the immature myocardium, but until now, no study reports whether LIP effects on endoplasmic reticulum stress apoptosis in immature myocardial cells. OBJECTIVE:To investigate the effect of LIP on endoplasmic reticulum stress and apoptosis. METHODS:Langendorff-perfused isolated rabbit hearts were used in this study. Twenty-four immature rabbits were randomized into three groups. Control group:Isolated rabbit heart was only perfused with Krebs-Henseleit for 180 minutes. I/R group:Isolated rabbit heart was perfused 20 minutes, and then ischemia for 60 minutes fol owed by reperfusion 100 minutes. LIP group:Limbs were repeatedly obstructed 5 minutes and relaxed 5 minutes for three times, to establish Langendorff models, and then repeated the method of ischemia/reperfusion in I/R group. The myocardial apoptosis was assayed with TUNEL method. The expression of glucose-regulated protein 78, Bcl-2, Bax and Fas was detected with western blot analysis. RESULTS AND CONCLUSION:Compared with I/R group, apoptosis rate was significantly lower, the expression of Bcl-2 was significantly higher, and the expression of glucose-regulated protein 78, Bax and Fas was significantly lower in LIP group. This study demonstrated that LIP regulates myocardial cellapoptosis through reducing the expression of endopasmic reticulum stress GRP78, Bax and Fas and increasing the expression of Bcl-2.
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There has been an ongoing search for clinically acceptable methods for the accurate, efficient and simple diagnosis and prognosis of hepatocellular carcinoma (HCC). Optical spectroscopy is a technique with potential clinical applications to diagnose cancer diseases. The purpose of this study was to obtain the optical properties of HCC tissues and non-tumorous hepatic tissues and identify the difference between them. A total of 55 tissue samples (HCC tissue, n=38; non-tumorous hepatic tissue, n=17) were surgically resected from patients with HCC. The optical parameters were measured in 10-nm steps using single-integrating-sphere system in the wavelength range of 400 to 1800 nm. It was found that the optical properties and their differences varied with the wavelength for the HCC tissue and the non-tumorous hepatic tissue in the entire wavelength range of research. The absorption coefficient of the HCC tissue (1.48±0.99, 1.46±0.88, 0.86±0.61, 2.15±0.53, 0.54±0.10, 0.79±0.15 mm(-1)) was significantly lower than that of the non-tumorous hepatic tissue (2.79±1.73, 3.13±1.47, 3.06±2.79, 2.57±0.55, 0.62±0.10, 0.93±0.16 mm(-1)) at wavelengths of 400, 410, 450, 1450, 1660 and 1800 nm, respectively (P<0.05). The reduced scattering coefficient of HCC tissue (5.28±1.70, 4.91±1.54, 1.26±0.35 mm(-1)) and non-tumorous hepatic tissue (8.14±3.70, 9.27±3.08, 2.55±0.57 mm(-1)) was significantly different at 460, 500 and 1800 nm respectively (P<0.05). These results show different pathologic liver tissues have different optical properties. It provides a better understanding of the relationship between optical parameters and physiological characteristics in human liver tissues. And it would be very useful for developing a non-invasive, real-time, simple and efficient way for medical management of HCC in the future.
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Objective To investigate the effect of oncolytic adenovirus vector SG600-IL24expressing human melanoma differentiation associated gene-7 (mda-7/IL-24) on hepatocellular carcinoma cell lines with different metastatic potential of HepG2, SMMC7721, MHCC97L and normal liver cell line LO2. Methods The oncolytic adenovirus SG600-IL24 which carrying mda-7/IL-24 gene was transfected into hepatocellular carcinoma cell lines and normal liver cell line. The mRNA and protein expression of mda7/IL-24 in HepG2, SMMC7721, MHCC97L and LO2 cell lines was confirmed by RT-PCR,ELISA assay and Western blot respectively. MTT assay and flow cytometry were used to study tumor cell proliferation and cell cycle in vitro. Hoechst33258 and flow cytometry were studied to indicate the apoptosis effects. Results It was confirmed by RT-PCR, ELISA assay and Western-blot that the exogenous mda-7/IL-24 gene was highly expressed in HepG2, SMMC7721, MHCC97L and LO2 cell lines. MTT and apoptosis detection indicated that MDA-7/IL-24 can induce the growth suppression (the inhibition rate was 75% ±2. 5% ,86% ±3. 5% ,and promotes apoptosis ( the apoptosis rate was 56. 5% ± 4. 0% , 34. 4% ± 2. 0% , 43. 3% ± 2. 5%cell lines at G2/M phase ( the blocking rate was 35. 4% ± 4. 2% , 40. 5% ± 5. 0% , 42. 0% ± 5. 0%metastatic potential hepatocellular carcinoma cell lines but not in normal liver cell line.Conclusions Oncolytic adenovirus vector SG600-IL24 can selectively induce growth suppression, promote apoptosis in hepatocellular carcinoma lines in vitro but not in normal liver cell LO2.
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Objective To explore the mechanism of melanoma differentiation associated gene-7(mda-7) in combination with adriamycin(ADM) killing the HCC HepG2 cells and reversing their multidrug resistance (MDR). Methods The experiment was conducted in three groups including the combined group, ADM group and mda-7 group. MTT assay and FCM were used to determine the differences among the 3 groups and clarify the reversing effect of combined treatment on multidrug resistance of the tumor cells. Expression levels of MDR-1, STAT-3, BCL-2, BAXmRNA were determined with real-time PCR. Western blotting was performed to observe the changes of proteins gp-l70, stat3,P-stat3, PKB, bcl-2,bax in all 3 groups. Result After transfection with 100VP/cell Ad. mda-7,the growth suppression rate of HepG2 treated by ADM (1.5 mg/L) rose from 17.46% to 79. 5%.According to the changes, killed HepG2 cells were increased by a factor of 4.55. times. MDR-1 mRNA was decreased from (16.49 ± 0. 11) to (5.48±0.05) and STAT-3 mRNA increased from (13.17±0. 08) to (21. 57±0. 11)(P＜0.05). Western blotting also showed that P-170 and PKB was decreased and the phosphorylation-stat-3 increased after the combined treatment. Conclusion Ad.mda-7 can reverse the multidrug resistance HepG2 cells. It inhibits the expression of MDR-1 mRNA,then arrests PKB protein and the signaling pathway of active stat-3 to induce apoptosis of HCC cells.
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Objective To determine the effect of nanochemotherapy drug on Survivin and p53 ex-pressed by human biliary traet carcinoma cell line QBC939.Methods Culturing the human biliary tract carcinoma cell line QBC939 in vitro and it was divided into five groups including saline,nanochemotherapy drug,nanopartiele withoul nanochemotherapy drug,5-FU and gemcitabine.Using the methods of MTT and flow cytometry to observe the growth of QBC939 which was dealt with different drugs.In addition,RT-PCR and Western blot were used to delect the expression of mRNA and protein by Survivin or p53.Results The expression of mRNA and protein by Survivin decreased in the following set:saline,nanoparlicle withoul nanochemotherapy drug,5-FU,gemcitabine and nanochemotherapy drug,respeclively.However,the ex-pression of mRNA and protein by p53 were in reverse order.The inhibiting action to QBC939 was obvious in nanochenmtherapy drng and the apoplotic rate was higher than others except for gemcitabine(P＜0.05). Conclusion Nanochemotherapy drug has significant effect on treatment cholangiocarcinoma in vilro,which may attribute to the down regulation of Survivin and up regulation of p53.
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The effect of targeted magnetic nanoparticles on hepatoma and the underlying mechanism were examined. Nude mice transplanted with a human hepatoma cell line (HepG2 cells) were randomized into 5 groups, including: (1) group A, receiving normal saline, (2) group B, receiving 5-fluorouracil (5-Fu), (3) group C, receiving magnetic nanoparticles containing 5-Fu, (4) group D, consisting of treatment with magnetic nanoparticles containing 5-Fu and inside magnetic field and (5) group E, receiving pure magnetic nanoparticles and inside magnetic field. Morphological features of transplanted tumors in mice in each group were observed under transmission electron microscope (TEM). The expression of bcl-2/bax protein was immunohistochemically detected by SABC method. The results showed that a large number of apoptotic tumor cells were found in group B and group D under TEM. The expression of bcl-2 protein was significantly decreased and the expression of bax protein increased significantly in both group B and D as compared with those in group A, C and E (P<0.01 for all). The decrease in bcl-2 and the increase in bax were more in group D as compared with group B (P<0.01). It is concluded that the targeted magnetic nanoparticles containing 5-Fu can improve the chemotherapeutic effect of 5-Fu by decreasing bcl-2 expression, increasing bax expression and inducing apoptosis of the liver cancer cells.
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Objective s To train postgraduate medical students the ability of effectively using network resources and independently studying,and to explore new model of clinical liver cancer teaching. Methods The teaching model of problem-based learning (PBL) to clinical liver cancer teaching was applied. Results The teaching model of PBL changed graduate student the status of passive acceptance to active participation. The teaching process was full of livingness,and the teaching quality was improved. Conclusion The teaching model of PBL can break through the limitations of passive acceptance of book knowledge in traditional teaching model and improve the ability to handle the comprehensive clinical knowledge of liver cancer,which provides a new model to the teaching of liver cancer to graduate medical students in clinic.
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Objective:To study the effect of recombined TNF ? combined with induction of apoptosis and its relation with expression of bcl 2 in SW480 human colon cancer cells.Methods:The inhibitive effect of 5 Fu and TNF ? was measured by MTT.Apoptosis of colon cancer cells was studied by TUNEL.The expression of bcl 2 gene was determined by immunohistochemical analysis.Results:After SW480 colon cancer cells were treated with combination in various concentration of 5 Fu and 3 000 U/mL of TNF ? for 48 h,MTT assay revealed a significant decrease of relative viability in compasrison to the control.When SW480 colon cancer cells were treated with 3 000 U/mL of TNF ? and 125 ug/mL of 5 Fu in combination for 48 h,TUNEL method showed the number of nick end positive cells was significantly increased in comparison to the control.The expression level of bcl 2 gene lowered significantly than that of the control group.Conclusion:These results confirmed that combined TNF ? and 5 Fu could synergistically induce apoptosis in SW480 cells and decrease the expression level of bcl 2 gene.
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Objective To study the effect of magnetic nanoparticles on human cholangiocarcinoma xenograft in nude mice, and compared with otherchemotherapy drugs Methods We established human cholangiocarcinoma xenograft in nude mice with QBC939 cell line.The nude mice were devided into 4 groups randomly.Saline,5-FU, Gemcitabine and magnetic nanoparticles were given to nude mice through tail vein on 20d after implanting QBC939 cell line. Calculations were done at different time after treatment in order to compare tumor volume,inhibition ratio of tumor and tumor growth curve of each group. The nude mice were killed on 35d after treatment to harvest tissue for electron microscopic examination to observe ultra-structural changes. Results The tumor volume of control, 5-FU, magnetic nanoparticles and Gemcitabine groups was (2256.1?267.1) mm3, (2096.5?237.9)mm3,(1392.2?189)mm3, and (1534.9?115 )mm3 respectively.The last two groups have significant difference compared to the first two groups(P