Research progress of autologous peritioneal patch for repairing defective blood vessels / 中华肝胆外科杂志

In the process of radical resection of abdominal malignant tumors, large blood vessels are often invaded, which not only increases the difficulty of operation, but also directly affects the curative effect and prognosis. As the concept of expanded radical surgery combined with vascular resection and reconstruction and related techniques have been gradually recognized, surgeons have begun to use autologous, allogeneic or artificial vessels to repair the defective blood vessels during surgery, so as to achieve the effect of radical surgery. However, due to the short comings of these materials, scholars have been looking for better alternatives. In view of the fact that the mesothelial cells of the peritoneum and the endothelial cells of the blood vessels have many similarities in embryology, histology and physiology, and peritoneum is also easier to obtain than autologous, allogeneic or artificial vessels. To make the autologous peritoneum into a patch for repairing the defective blood vessels is feasible in theoretical and technical. In this paper, we review the current research progress of autologous peritoneal patch to repair blood vessels of defect.

Fundamental pathological mechanisms underlying gastro-intestinal cancer peritoneal metastasis / 中华胃肠外科杂志

Gastrointestinal cancer peritoneal metastasis(GICPM) is one of the biggest challenges of clinical treatment. The ultimate solution to the problem requires the clinicians to accurately understand cytologic and molecular pathological mechanisms behind GICPM, and apply such knowledge in the clinical decision-making process for diagnosis and treatment of individual patient, so as to realize "prevention" and "treatment" proactively. The core cytopathological mechanisms behind GICPM, which are closely related to clinical treatment decisions, are as follows: (1) free cancer cells or clusters in peritoneal cavity colonize the peritoneum, resulting in irreversible pathological damage to peritoneal mesothelial cells; (2) the colonized cancer cells further invade the specific structure of the peritoneal milky spots and initiate an accelerated invasive growth process; (3) the process of peritoneal interstitial fibrosis aggravates the structural destruction of the peritoneum; (4) the interaction between cancer cells and immune cells in the milk spots forms a permissive immune microenvironment that promotes the growth of peritoneal metastatic cancer. These four core cytopathological mechanisms are mutually causal and promote each other, forming a vicious circle of GICPM development. As long as clinicians accurately understand these four points, it is possible to grasp the opportunity of clinical diagnosis and treatment, change reactive and passive treatment into preventive and proactive treatment, and improve the clinical diagnosis and treatment landscape of GICPM.

Effect of Banxia Xiexintang-containing Serum on Epithelial-mesenchymal Transition of Peritoneal Mesothelial Cells Induced by Gastric Cancer-derived Exosomes / 中国实验方剂学杂志

Objective:To investigate the effect of Banxia Xiexintang on the epithelial-mesenchymal transition （EMT） of human peritoneal mesothelial cell line （HMrSV5） induced by gastric cancer-derived exosomes （Exo）. Method:Banxia Xiexintang-containing serum was prepared and the human gastric cancer NCI-N87-derived exosomes （NCI-N87-Exo） were extracted， followed by their identification by transmission electron microscopy and Western blotting and labeling with 1，1-dioctadecyl-3，3，3，3- tetramethylindocarbocyanine perchlorate （Dil）. The cells were divided into the blank group， model group， and low-， medium-， and high-dose （13.5，27，54 g·kg^-1） Banxia Xiexintang groups. HMrSV5 cells in the blank group were cultured alone， the ones in the model group with 100 mg·L^-1 NCI-N87-Exo， and those in the low-， medium-， and high-dose Banxia Xiexintang groups with 100 mg·L^-1 NCI-N87-Exo plus low-， medium-， and high-dose 10% Banxia Xiexintang-containing serum， respectively. Confocal laser microscope was used to observe the uptake of NCI-N87-Exo by HMrSV5 cells at 24 h， 48 h and 72 h. Seventy-two hours later， the morphological changes in HMrSV5 cells were observed. The protein expression levels of E-cadherin， cytokeratin 19 （CK19）， <italic>α</italic>-smooth muscle actin （<italic>α</italic>-SMA）， elastin， and transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁）， Smad2/3， and p-Smad2/3 were assayed by Western blot. Result:It was observed under the transmission electron microscope that NCI-N87-Exo showed an oval or dish-shaped vesicle structure with a particle size ranging from 40 to 80 nm. Exo marker proteins CD9 and CD63 were highly expressed while calreticulin was not expressed， implying that the NCI-N87-Exo was confirmed. After 24 h， 48 h， 72 h of co-culture， it was observed under the fluorescence microscope that NCI-N87-Exo were taken up by HMrSV5 cells， which was positively correlated with time. Compared with the blank group， Banxia Xiexintang significantly inhibited the uptake of NCI-N87-Exo by HMrSV5 cells， with better effect noticed in the middle- and high-dose Banxia Xiexintang groups（<italic>P</italic><0.05，<italic>P</italic><0.01）. After intervention with Banxia Xiexintang-containing serum， the HMrSV5 cells were arranged densely， and the intercellular space was significantly reduced， with the most obvious changes present in the high-dose Banxia Xiexintang group. Western blot revealed that the protein expression levels of E-cadherin and CK19 in HMrSV5 cells after being intervened with the medium- and high-dose Banxia Xiexintang-containing serum were increased significantly as compared with those in the blank group， whereas the levels of <italic>α</italic>-SMA and Elastin were decreased significantly （<italic>P</italic><0.01）. Banxia Xiexintang-containing serum at the low， medium， and high doses remarkably down-regulated TGF-<italic>β</italic>₁ and p-Smad2/3 protein expression（<italic>P</italic><0.05，<italic>P</italic><0.01）. However， there was no significant change in Smad2/3. Conclusion:NCI-N87-Exo can be taken up by HMrSV5 cells to induce EMT. Banxia Xiexintang can inhibit the uptake of NCI-N87-Exo by HMrSV5 cells and the resulting EMT induced by NCI-N87-Exo， which is related to the regulation of TGF-<italic>β</italic>₁/Smads signaling pathway.

Effect of KLF-4 on peritoneal fibrosis induced by high glucose peritoneal dialysate via targeting regulation of the expression of E-cadherin / 解放军医学杂志

[Abstract] Objective To explore the relationship between Kruppel-like factor 4 (KLF-4) and E-cadherin in human peritoneal mesothelial cells (HPMCs), and the expression and function of KLF-4 in the animal model of peritoneal fibrosis induced by high glucose peritoneal dialysate. Methods Co-transfection in HPMCs with the plasmid of KLF-4 and the bind site or mutant in the promoter region of E-cadherin, and then the luciferase activity was measured of the each bind site and its matched mutants to estimate whether KLF-4 can combine with the bind site in the promoter region of E-cadherin; Chromatin immunocoprecipitation (CHIP) was exploited to verify if KLF-4 can combine with the bind site in the promoter region of E-cadherin; Real-time PCR and Western blotting were performed to detect the expression of E-cadherin at the bind site and matched mutants of b, d, f and g. Thirty SD rats were randomly divided into saline group, peritoneal dialysate group and experimental group (10 each). Rats in saline group were given intraperitoneal injection with 0.9% NaCl, in peritoneal dialysate group were given with 4.25% high glucose peritoneal dialysate, and in experimental group were given via tail vein with 4.25% high glucose peritoneal dialysate and the mixture of KLF-4 plasmid suspension containing ultrasound microbubble. To observe the peritoneal tissue thickness of the 3 groups of rats by Hematoxylin and Eosin staining. Masson trichrome staining was performed to detect the deposition of collagen fibers in peritoneal tissue of the 3 groups of rats. Immunohistochemistry was used to detect the expression level of KLF-4, E-cadherin, α-SMA and fibronectin (FN) in peritoneal tissue of the 3 groups of rats. Results Promoter luciferase reporter gene and CHIP results showed that KLF-4 can combine with the bind site in the promoter region of E-cadherin in HPMCs. Real-time PCR and Western blotting showed that KLF-4 can positively regulate the expression of E-cadherin. HE staining showed that the peritoneal tissue was obviously thickened in rats of peritoneal dialysate group [(105.91±12.0) μm] than in rats of saline group [(20.89±5.39) μm] and of experimental group [(23.05±6.07) μm] with statistical significance (P0.05). Masson staining showed that the deposition of collagen fiber significantly increased in peritoneal dialysate group (0.89±0.09) than in saline group (0.19±0.03) and experimental group (0.15±0.06) with statistical significance (P0.05). Immunohistochemistry results showed that the expressions of KLF-4 and E-cadherin were obviously lower in peritoneal dialysate group (0.27±0.09, 0.31±0.03) than in saline group (0.79±0.19, 0.83±0.13) and experimental group (0.85±0.11, 0.76±0.11) with statistically significant difference (P0.05). In contrast, the expressions of α-SMA and FN were evidently higher in peritoneal dialysate group (0.83±0.09, 0.63±0.09) than in saline group (0.22±0.08, 0.30±0.07) and experimental group (0.19±0.05, 0.11±0.03) with statistically significant difference (P0.05). Conclusion KLF-4 may positive regulate the expression of E-cadherin by combining with the bind site in the promoter region of E-cadherin, and inhibit the peritoneal fibrosis induced via high glucose peritoneal dialysate.

Evaluation of Effusion Fluid (Pleural and Peritoneal) with Special Reference To Cell Block Preparation and Immunohistochemistry for Differentiating Between Reactive Mesothelial and Malignant Effusion

Background: The serous cavities are lined by a single layer offlat mesothelial cells called the serosa. Normally these cavitiesare collapsed and contain only a small amount of fluid, enoughto lubricate the adjacent surfaces. Cytological examination ofserous fluid is of paramount importance. It reveals informationabout inflammatory conditions of serous membrane, infectionby bacteria, fungi, virus and presence of malignant cells.Differentiation of population of reactive mesothelial cells fromthose of malignant cells remains a diagnostic challenge inconventional cytological smear. To overcome this challenge,cell block technique along with immunocytochemistry gives abetter histoarchitectural pattern and support immensely forcategorising the effusion to be reactive or malignant.Aims and Objectives: To evaluate utility of cell blocktechnique in effusion fluid (pleural and peritoneal) using limitedimmunohistochemistry markers for differentiating betweenreactive mesothelial and malignant mesothelial cells.Materials and Methods: The present study was carried out inDepartment of Pathology at M.K.C.G MCH, Berhampur,Odisha over a time period from July 2016- July 2018. A total of90 cases were evaluated. The fluids were stained with routinecytological stains. Cases on evaluation of cytomorphology ifsuspicious for malignancy, cell block was prepared. Cell blockwere stained both for routine hematoxylin and eosin andimmunohistochemistry with EMA (Epithelial marker antigen) forepithelial cells and Calretinin for mesothelial cells.Results: A total of 90 cases were evaluated cytologically. 40cases showed benign features and 24 cases showedmalignant features on cytomorphology alone. 26 cases weresuspicious for malignancy which on cell block preparation andimmunocytochemistry were differentiated as benign (10 cases)or malignant (16 cases). EMA showed 97.5 % sensitivity and98% specificity. Calretinin showed 100 % sensitivity and 97.5%specificity.Conclusion: The use of cytopathology of pleural andperitoneal effusion is helpful for early diagnosis and treatment.The technique is cheap, easy to perform and produces speedydiagnosis. In the identification of malignant cells in effusion andits differentiation from cells showing reactive and degenerativechanges there were diagnostic difficulties in some of the cases.Immunocytochemistry is an important diagnostic tool ineffusion cytology.

Utility of MOC-31 monoclonal antibody in differentiating metastatic adenocarcinoma cells and reactive mesothelial cells in effusion cytology

In effusion cytology, a clear distinction between reactive mesothelial cells and metastatic adenocarcinoma cells is sometimes challenging mainly due to similarities in the cytomorphological features. In such cases for definitive diagnosis, paraffin-embedded cell block examination and immunohistochemistry are helpful in making this distinction. MOC-31 is one of the proposed immunomarker for adenocarcinoma cells. We undertook to evaluate the role of MOC-31 as a marker for identifying adenocarcinoma cells in effusion specimen. A total of 185 paraffin-embedded cell blocks of effusion samples were identified, of these 111 cases were of metastatic adenocarcinoma. MOC-31 was positive in 101 of the 111 cases of metastatic adenocarcinoma. Minimal focal cytoplasmic staining was also seen in 7 of the 74 cases of reactive mesothelial cells, but these were taken negative as they did not show membrane positivity. The sensitivity and specificity of MOC-31 for metastatic adenocarcinoma cells were 92.5%, and 100% respectively, positive and negative predictive value (NPV) was 100% and 91.14%, respectively. MOC-31 can be used as a reliable marker in effusions for distinguishing metastatic adenocarcinoma from reactive mesothelial cases.

Effect of lnc-MGC on the trans-differentiation of peritoneal mesothelial cells induced by high glucose / 解放军医学杂志

Objective To explore the role of long noncoding RNA-MGC (lnc-MGC) on the trans-differentiation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods The immortalized HPMCs were used to establish control group and high glucose group (60mmol/L) respectively.Cells in control group were cultured with ordinary cell medium,and in high glucose group were stimulated with high glucose medium for 72h.Changes of lnc-MGC expression in the both groups were measured by RT-PCR,and the changes of mRNA and protein expression of E-Cadherin,α smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),type Ⅰ collagen (COL-1) and type Ⅲ collagen (COL-3) in epithelial cells of both groups were measured by RT-PCR and Western blotting.The HPMCs were transfected with lentivirus,and then the changes of the above indexes were observed after up-and down-regulation of lnc-MGC.Results By high glucose stimulation of HPMCs for 72h,RTPCR results showed that the expressions oflnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA increased obviously (P＜0.05),and the expression of E-Cadherin mRNA decreased markedly (P＜0.05) in high glucose group than in control group;Western-blotting results indicated that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).After lentivirus transfection and down-regulation oflnc-MGC,RT-PCR results showed that the expressions of lnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA decreased obviously (P＜0.05),and the expression of E-Cadherin mRNA increased markedly (P＜0.05) in high glucose group than in control group;Western-blotting results showed that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).After lentivirus transfection and upregulation of lnc-MGC,RT-PCR results showed that the expressions of lnc-MGC,α-SMA,CTGF,COL-1 and COL-3 mRNA increased significantly (P＜0.05),and the expression of E-Cadherin mRNA decreased obviously (P＜0.05) in high glucose group than in control group;Western blotting results showed that the expression of protein was consistent with that of mRNA,and the differences were statistically significant (P＜0.05).The downstream target was predicted as miRNA126-3p,and compared with the control group,the expression ofmiRNA126-3p increased (P＜0.05) after high glucose stimulation,and after transfection with down regulated lnc-MGC lentivirus,the expression of miRNA126-3p decreased obviously (P＜0.05),and transfection with up regulated lnc-MGC lentivirus,the expression ofmiRNA126-3p increased obviously (P＜0.05).Conclusions lnc-MGC participates in the process of HPMCs transdifferentiation through regulating miRNA126-3p.Regulation oflnc-MGC expression level may control the phenotype transition of HPMCs,and delay the development of peritoneal fibrosis.

Effects of protein PUMA-α on the apoptosis and fibrosis of human peritoneal mesothelial cells induced by high glucose / 解放军医学杂志

Objective To explore the effects of p53 up-regulated modulator of apoptosis (PUMA)-α protein on the apoptosis and fibrosis of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs were induced by 50mmol/L D type glucose or mannitol for 72 hours respectively,flow cytometry was employed to detect the rate of apoptotic cells,and theexpression levels of apoptosis-and fibrosis-related proteins were detected by Western blotting.The untreated HPMCs were transfected with Lenti-PUMA-α,and the treated cells were transfected with shRNA-PUMA-α,the number of apoptotic cells and the expression levels of apoptosis-and fibrosis-related proteins were detected with the methods mentioned above.Results Flow cytometry showed that the rate of apoptotic HPMCs increased after being induced by high glucose for 72 hours,and Western blotting revealed that the expression levels of pro-apoptotic and pro-fibrotic related proteins increased,but the arrestins of apoptosis and fibrosis-related proteins decreased.Up-regulation of PUMA-α promoted apoptosis and fibrosis,while down-regulation of PUMA-α alleviated apoptosis and fibrosis of HPMCs.Conclusion High glucose may accelerate apoptosis and fibrosis of HPMCs by up-regulating the expression of PUMA-α.

Effect of high glucose-based peritoneal dialysis fluids on NLRP3-IL-1β in human peritoneal mesothelial cells / 北京大学学报(医学版)

Objective:To explore the effect of high glucose-based peritoneal dialysis fluids on NLRP3-IL-1β in human peritoneal mesothelial cells.Methods:HMrSV5 cells (SV40 immortalized human peritoneal mesothelial cell line) were grown in type Ⅰ collagen-coated dishes in DMEM/F12 containing 10％ fetal calf serum (FCS).All experiments on HMrSV5 cells were performed between passages 5 and 10.The cells were divided into 7 groups:control,1.5％ dextrose,2.5％ dextrose,4.25％ dextrose,rotenone,thenoyltrifluoroacetone (TTFA),and antimycin A.Immunoblotting was used to evaluate the expression of IL-1 β.Small interfering RNA (siRNA) targeting NLRP3 was used to downregulate the expression of NLRP3 and Western blot was used to evaluate the expression of IL-1 β in human peritoneal mesothelial cells exposed to 4.25％ dextrose.In the meanwhile,resveratrol (RSV) was used to induce autophagy,3-methyladenine (3-MA) and siRNA against Beclin 1 or ATG5 were used to block autophagy,flow cytometric was used to analyze the respiring (mitotracker deep red),total (mitotracker green) and reactive oxygen species (ROS)-generating mitochondria (mitoSOX);Western blot was used to evaluate the expression of IL-1β.Results:The IL-1β relative expressions were 0,0.175 ±0.082,0.418 ± 0.163,2.357 ±0.288,2.642 ±0.358,3.271 ±0.462,and 0.123 ±0.091,indicating that the cells exposed to high glucose-based peritoneal dialysis fluids and cells treated with mitochondria respiratory chain key enzyme complex Ⅰ,and complex Ⅲ inhibitors increased the IL-1β expression.And we found that NLRP3 knock-down significantly blocked the upregulation of IL-1 β.In addition,the fluorescence intensity of total mitochondria and ROS-generating mitochondria in the following groups:control,negative control,RSV,3-MA,ATG5 siRNA,Beclin1 siRNA were 1.76 ± 0.42,1.83 ± 0.55,1.85 ± 0.62,7.36 ± 0.92,5.35 ± 0.77,5.06 ± 0.62 and 821.68 ± 95.12,868.15 ± 102.82,723.39 ± 92.56,1 660.08 ± 113.65,1 433.01 ± 107.24,1 562.36 ± 112.88 respectively.The increased concentrations of mitochondrial ROS and IL-1β upregulation were confirmed in the inhibition but not the induction of autophagy.We also found that downregulation of ATG5 and Beclin1 sensitized cells for the release of IL-1β induced by MSU (monosodium urate) or nigericin which was the NLRP3 inflammasome activator.RSV treatment attentuated this effect.Conclusion:Long-term application of high glucose-based peritoneal dialysis fluids can trigger the consistent activation of NLRP3-IL-1ββ in peritoneal mesothelial cells.Timely initiation of autophagy may block the NLRP3-IL-1ββ activation and provide a basis for the further development of a potential therapeutic strategy for delay of chronic inflammation and peritoneal fibrosis associated with peritoneal dialysis.

Effects of LncRNA-ATB on human peritoneal mesothelial cells phenotypic transition and proliferation induced by high glucose / 解放军医学杂志

Objective To explore the effect of long noncoding RNA-ATB (LncRNA-ATB) on phenotypic transition and proliferation of human peritoneal mesothelial cells (HPMCs) induced by high glucose.Methods HPMCs used in experiment were divided into three groups:control group,mannitol group and hypertonic glucose group.HPMCs in control group received no treatment,and in hypertonic glucose group and mannitol group were treated with 50mmol/L D-glucose and isotonic mannitol for 72 hours,respectively.Real-time PCR was employed to detect the mRNA expression of LncRNA-ATB,E-cadherin,α-smooth muscle actin (α-SMA),connective tissue growth factor (CTGF),Cyclin D1,cyclin dependent kinase inhibitor 4 (CDK4),protein 27 (p27)and proliferating cell nuclear antigen (PCNA).Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA,CTGF,Cyclin D1,CDK4,p27 and PCNA,and flow cytometry was used to test the cell cycle.Lentivirus artifice was used to up-or down-regulate the expression of LncRNA-ATB in untreated HPMCs.Real-time PCR was employed to detect the mRNA expression of E-cadherin,α-SMA and CTGF,Western blotting was performed to detect the proteins expression of E-cadherin,α-SMA and CTGF,and flow cytometry was used to test the cell cycle.Results It is revealed by Real-time PCR,Western blotting and flow cytometry that the expressions increased of LncRNA-ATB,α-SMA,CTGF,Cyclin D1,CDK4 and PCNA induced by hypertonic glucose,and decreased of E-cadherin and p27 (P＜0.05).Up-regulation of LncRNA-ATB promoted HPMCs phenotypic transition and proliferation,while down-regulation alleviated HPMCs phenotypic transition and proliferation.Conclusion Hypertonic glucose may accelerate HPMCs phenotypic transition and proliferation by up-regulating the expression of LncRNA-ATB.

Danhong Injection inhibits high glucose-induced injury in rat peritoneal mesothelial cells / 中成药

AIM To investigate that Danhong Injection may decrease inflammation and oxidative stress response in rat peritoneal mesothelial cells (RPMCs) cultured with high glucose.METHODS The third passage of RPMCs were used for the experiment.After being incubated with DMEM for 24 hours,RPMCs were divided into normal control group,high glucose group (RPMCs were incubate with 100 mmol/L glucose for 12 h,24 h,48 h,72 h),Danhong Injection group (complete medium with 80 mL/L Danhong Injection for 48 h) and high glucose + Danhong Injection group (100 mmol/L high glucose with 40 mL/L,80 mL/L,160 mL/L Danhong Injection for 48 h respectively).The cell proliferation was detected by MTT method.The levels of interleukin-18 (IL-18),interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in culture fluid were determined by enzyme linked immunosorbent assay.The contents of malondialdehyde (MDA),superoxide dismutase (SOD),glutathione (GSH),in the culture fluid and intracellular reactive oxygen species (ROS) were measured by kits.The expressions of Bcl-2,Bax,fibronectin (FN),endothelin-1 (EDN1) and haem oxygenase-1 (HMOX1) mRNA were detected by quantitative real-time PCR.The protein expressions of EDN1 and HMOX1 were measured by Western blot.RESULTS High glucose can significantly inhibit RPMCs' viability.High glucose can also up-regulate the expressions of IL-18,IL-6,TNF-α,ROS,MDA,Bax mRNA,FN mRNA,EDN1 mRNA and down-regulate the levels of SOD,GSH,Bcl-2 mRNA,HMOX1 mRNA in RPMCs.Treatment with Danhong Injection can effectively reverse aforementioned effect induced by high glucose.CONCLUSION Danhong Injection can reverse the inhibition of cell viability,the inflammation and oxidative stress response induced by high glucose in RPMCs,thus protect peritoneal membrane.

Protective effect and its mechanism of Shenkang injection on peritoneal mesothelial cells in CAPD mice / 天津医药

Objective To study the protective effect of Shenkang injection on peritoneal mesothelial cells (PMCs) of continuous ambulatory peritoneal dialysis (CAPD) mice, and explore the possible mechanism. Methods Forty ICR mice were randomly divided into blank control (A) group, peritoneal dialysis (B) group, low dose of Shenkang (C) group ( 2.5%dialysate+5 mL/kg Shenkang injection), medium (D) group (2.5% dialysate+10 mL/kg Shenkang injection) and high (E) group (2.5% dialysate+ 20 mL/kg Shenkang injection). Mice were observed for 4 weeks. Fasting blood glucose, total cholesterol, triglyceride and C-reactive protein (CRP) were detected by biochemical assay. Tumor necrosis factor-alpha (TNF-α), transforming growth factor-beta 1 (TGF-β1), vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF) levels of serum and dialysate were detected by ELISA. Pathological changes of peritoneal tissue were observed by HE staining. Expression and mRNA transcription levels of these four cytokines in the peritoneal tissue were detected by immunohistochemical staining and real-time PCR respectively. Results There were no significant differences in body weight, fasting blood glucose, total cholesterol and triglyceride between 5 groups of mice (P>0.05). Compared with group B, there was no significant difference in CRP level between group C and group E, but which was significantly decreased in group D (P<0.05). The serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were decreased in group C and group D. The serum and dialysate levels of TNF-αand TGF-β1 were significantly increased in group E (P<0.05), but there was no significant difference between VEGF and CTGF in group E. Compared with group E, except for CTGF in dialysate of group C, the serum and dialysate levels of TNF-α, TGF-β1, VEGF and CTGF were significant decreased in group C and group D (P<0.05). Damaged PMCs were found in group B, which were improved in various degrees in group C, group D and group E. Compared with group B, the protein expression and mRNA relative transcription levels of TNF-α, TGF-β1, VEGF and CTGF tended to decrease gradually in group C, group D and group E (P<0.05). Conclusion A certain concentration of Shenkang injection can protect PMCs by inhibiting the expression of TNF-α, TGF-β1, VEGF and CTGF in CAPD mice, so as to control the occurrence and development of peritoneal fibrosis.

The influence of high glucose on cytokine secreted by peritoneal mesothelial cells in rats and the intervention effect of matrine / 重庆医学

Objective This study was designed to investigate the effect of matrine on high glucose‐induced tumor necrosis factor‐α(TNF‐α)and transforming growth factor‐β1 (TGF‐β1 ) overexpression in rat peritoneal mesothelial cells(PMCs) ,and to ex‐plore the possible mechanism in the peritoneal fibrosis .Methods ology The rat PMCs were incubated in vitro ,and then assigned in‐to 5 groups on the basis of the added drug concentrations :2 .5% glucose group (n=15 ,A);Glucose + 2 .5% matrine group (n=15 ,B);4 .25% glucose group (n=15 ,C);Glucose + 4 .25% matrine group (n=15 ,D);Blank control group (n=15 ,control group , added the DMEM/F12 medium) .The levels of TNF‐αand TGF‐β1 in the supernatants were measured by ELISA method at the dif‐ferent time (24h ,48h ,72h) ,and observed the changes of PMCs under the microscope .Results Compared with the groups of A ,C and D ,the PMCs in the control group and B group were small ,round ,spindle ,irregular or intensive .The expression of TNF‐αand TGF‐β1 in the A group was lightly higher than that of the control group ,but the level in the C group was significantly higher than that of the control group (P 0 .05 ) ,but the lever in the D group was significantly lower than that of the C group ( P< 0 .05 ) . Conclusion Matrine might be helpful to resist the overexpression of TNF‐αand TGF‐β1 induced by high glucose ,and then protect the PMCs .

Effect of ligustrazine nanoparticles nano spray on transforming growth factor-β/Smad signal pathway of rat peritoneal mesothelial cells induced by tumor necrosis factor-α / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of ligustrazine nanoparticles nano spray (LNNS) on transforming growth factor β (TGF-β)/Smad signal protein of rat peritoneal mesothelial cells (RPMC) induced by tumor necrosis factor α (TNF-α), and the anti-adhesion mechanism of LNNS in the abdominal cavity.</p><p><b>METHODS</b>The primary culture and subculture of rat peritoneal mesothelial cells (RPMC) was processed by trypsin digestion method in vitro. The third generation was identifified for experiment and divided into 5 groups: a blank group: RPMC without treatment; a control group: RPMC stimulated with TNF-α; RPMC treated by a low-dosage LNNS group (2.5 mg/L); RPMC treated by a medium-dosage LNNS group (5 mg/L); and RPMC treated by a high-dosage LNNS group (10 mg/L). Reverse transcription-polymerase chain reaction was applied to test the expression of fifibronectin, collagen I (COL-I), TGF-β mRNA, and Western blot method to test the Smad protein 7 expression of RPMC.</p><p><b>RESULTS</b>Compared with the blank group, a signifificant elevation in fifibronectin (FN), COL-I and TGF-β mRNA expression of RPMC were observed in the control group (P<0.05). Compared with the control group, LNNS suppressed the expressions of FN, COL-I and TGF-β mRNA in a concentrationdependent manner (P<0.05). The expression of Smad7 protein of RPMC was down-regulated by TNF-α stimulation, and up-regulated with the increase of LNNS dose (P<0.05).</p><p><b>CONCLUSIONS</b>TNF-α may induce changes in RPMC's viability, leading to peritoneal injury. LNNS could reverse the induction of fifibrosis related cytokine FN, COL-I and TGF-β, up-regulating the expression of Smad7 by TNF-α in RPMC, thus attenuate peritoneal injury by repairing mesothelial cells.</p>

Effect of peritoneal dialysis fluids on the expression of TLR2 and TLR4 on peritoneal mesothelial cells / 重庆医学

Objective To investigate the effect of glucose-based peritoneal dialysis fluids and icodextrin-based peritoneal dial-ysis fluids on the expression of TLR2 and TLR4 on huamn peritoneal mesothelial cells .Methods Human peritoneal mesothelial cell line 5 - 10 generations(HMrSV5) was cultured in DMEM /F12 medium supplemented with 10% (v/v) fetal calf serum (FCS) .Cell viability and cell proliferation were assessed using M TT method .The experiment were divided into 5 different groups :group A (control group) ,1 .5% dextrose group ,2 .5% dextrose group ,4 .25% dextrose group and 7 .5% Lcodextrin group .Icodextrin group (aikau dextrin) ,TLR2 and TLR4 expression were detected by Western blot .Results Treatment with different concentrations of glucose-based peritoneal dialysis fluids for 24 h did not affect the expression of TLR2 and TLR4 protein .In addition ,after stimula-tion for 48 h ,1 .5% dextrose ,2 .5% dextrose ,4 .25% dextrose decreased TLR2 expression by (5 .5 ± 2 .8)% ,(31 .4 ± 7 .5)% , (54 .9 ± 1 .9)% respectively ,TLR4 expression by (32 .9 ± 17 .6)% ,(47 .7 ± 13 .5)% ,(66 .4 ± 13 .5)% respectively .Stimulation for 72 h ,decreased TLR2 expression by (29 .4 ± 14 .7)% ,(38 .9 ± 9 .9)% ,(63 .5 ± 16 .5)% respectively ,TLR4 expression by(59 .5 ± 16 .8)% ,(63 .1 ± 9 .5)% ,(79 .2 ± 14 .0)% respectively .There was no significant change in TLR2 and TLR4 protein expression on 7 .5% icodextrin group .Conclusion Glucose-based peritoneal dialysis fluids ,but not icodextrin-based peritoneal dialysis fluids downregulates expression of TLR2 and TLR4 by HM rSV5 .

The role of SRF- miRNA-143-KLF-4 signal pathway in the HPMC phenotypic transition induced via high glucose / 解放军医学杂志

Objective To explore the role of Kruppel-like factor 4 (KLF-4) in phenotypic transition of human peritoneal mesothelial cells (HPMCs) induced via high glucose. Methods HPMCs were induced by 50mmol/L glucose for 72 hours, the expressions of epithelium-cadherin (E-cadherin), KLF-4, α-smooth muscle actin (-SMA), connective tissue growth factor (CTGF) and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. The treated cells were transfected with LVKLF-4 and inhibitor, the untreated cells were transfected with shRNA-KLF-4 and mimic. The mRNA and protein expressions of KLF-4, E-cadherin, α-SMA, CTGF and miRNA-143 were detected by Real-time PCR and Western blotting, respectively. Results Real-time PCR showed that the expression of E-cadherin decreased and of α-SMA, CTGF and miRNA-143 increased, but of KLF-4 not changed in high glucose treated cells. Western blotting showed that the expression of KLF-4 and E-cadherin decreased. Upregulating KLF-4 increased the expression of E-cadherin, but decreased the expression of α-SMA and CTGF. Down-regulating KLF-4 decreased the expression of E-cadherin, but augment the expression of α-SMA and CTGF. Conclusion High glucose may induce the down-regulation of KLF-4 protein, and SRF- miRNA-143-KLF-4 signal pathway axis may be involved in the process of HPMC phenotypic transition.

Benign cystic peritoneal mesothelioma: an abdominal daze.

Benign cystic peritoneal mesothelioma (BCPM) is a rare clinical entity with varied presentations. Its obscure etiology and association with various intraabdominal conditions makes a precise diagnosis difficult. Diagnostic accuracy and diligent follow-up are essential because, though benign nature, it recurs locally. Herewith reporting a case of BCPM presenting as acute abdomen.

Effect of AstragalosideⅣ on Expression of High Glucose Dialysis Solution-Induced Fibrosis Cytokines in Human Peritoneal Mesothelial Cells / 中国中医药信息杂志

Objective To investigate the effects of AstragalosideⅣ (AST) on the expression of high glucose peritoneal dialysis solution (PDS)-induced transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF) in cultured human peritoneal mesothelial cells. To discuss the regulating effect of AST on induced fibrosis cytokines.Methods The HMrSV5 cell line was cultivated to the fifth generation and divided into normal group (10%FBS cultivation solution), model group (4.25% PDS and 10% FBS cultivation solution) and the low, medium and high doses AST groups (4.25% PDS with a respective 10, 20, 40 μg/mL AST). MTT colorimetric assay was employed to detect cell activity and ELISA was applied to detect expression of TGF-β1, CTGF and VEGF in cultured supernatants.Results Except for 5 μg/mL group, HPMCs activity of high glucose plus different concentration AST groups were enhanced in different degrees, especially with 40, 45, and 50 μg/mL (P<0.05). The expression of TGF-β1, CTGF, and VEGF in model group increased. Compared with the control group, expression of the three AST groups significantly decreased and showed dose-effect relationship (P<0.05). Conclusion AST could reduce the expression of TGF-β1, CTGF and VEGF in high glucose-induced HPMCs and was useful in slowing down the progress of peritoneal fibrosis.

PI3K/Akt signaling regulates epithelial-mesenchymal transition of peritoneal mesothelial cells in peritoneal dialysis / 中华肾脏病杂志

Objective To investigate the role of PI3K/Akt signaling in the regulation of epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) in peritoneal dialysis in vitro and in vivo.Methods The level of Phosphorylated serine/threonine kinase Akt and the expression of EMT associated gene and protein,including ZO-1,Vimentin and FN,were measured in mice EMT model.In vitro study,phosphorylation level and nuclear translocation of Akt,ZO-1 and Vimentin expression induced by TGF-β1 in human peritoneal mesothelial cells (HPMCs) were also observed.Moreover,HPMCs were pre-treated by one of PI3K/Akt inhibitor,LY294002,or transfected with dominant-negative Akt plasmid to inhibit PI3K/Akt signaling,then analyzed its effect on Zo-1 and Vimentin expression induced by TGF-β1.Results Compared with the control,thickened parietal peritoneum and remarkable decrease in mRNA and protein of the epithelial marker ZO-1,and notable increased in the expression of mesenchymal markers Vimentin and FN were observed in PD mice (all P ＜ 0.01).Moreover,the phosphorylation of Akt also significantly increased under above condition (P ＜ 0.01).In vitro study,with the stimulation of TGF-β1,the expression of Zo-l was down-regulated,while the expression of Vimentin increased (all P ＜ 0.01).In addition,TGF-β1 remarkably increased pAkt in HPMCs (all P ＜ 0.01) in dose-dependent.However,LY294002 and DN-Akt dramatically inhibited the vimentin expression in HPMCs induced by TGF-β1 after inhibition of pAkt.On the other hand,the expression of ZO-1 also was restored (P ＜ 0.01).Conclusion PI3K/Akt signaling is involved in EMT of peritoneal mesothelial cells in peritoneal dialysis,and may be a new target for the prevention and treatment of peritoneal fibrosis during PD.

Expression of CD44 in endometrial stromal cells from women with and without endometriosis and its effect on the adherence to peritoneal mesothelial cells

OBJECTIVE: This study was performed to compare the expression of CD44 in endometrial stromal cells (ESCs) of women with and without endometriosis and to evaluate the role of CD44 in the adherence of ESCs to peritoneal mesothelial cells (PMCs). METHODS: A PMC adherence assay was performed to evaluate the adherence of ESCs to PMCs in women with and without endometriosis. The expression of CD44 mRNA was measured by reverse transcription-polymerase chain reaction. CD44 protein was evaluated by Western blot analysis. RESULTS: There were no significant differences in the expression of CD44 mRNA and protein in ESCs or in the binding of ESCs to PMCs between patients with endometriosis and controls. Although the expression of CD44 protein was decreased in both women with endometriosis and controls after anti-CD44 antibody treatment, there was no effect on binding of ESCs to PMCs. Treatment of ESCs with peritoneal fluid from endometriosis patients resulted in a significant increase in binding of ESCs to PMCs compared to untreated ESCs in the endometriosis group. CONCLUSION: This study demonstrates that the expression of CD44 protein in ESCs from women with endometriosis might not be directly associated with adherence to PMCs.