Prospective application of stem cells to prevent post‐operative skeletal fibrosis

Post‐operative skeletal fibrosis is considered one of the major complications causing dysfunction of the skeletal system and compromising the outcomes of clinical treatment. Limited success has been achieved using current therapies; more effective therapies to reduce post‐operative skeletal fibrosis are needed. Stem cells possess the ability to repair and regenerate damaged tissue. Numerous studies show that stem cells serve as a promising therapeutic approach for fibrotic diseases in tissues other than the skeletal system by inhibiting the inflammatory response and secreting favorable cytokines through activating specific signaling pathways, acting as so‐called medicinal signaling cells. In this review, current therapies are summarized for post‐operative skeletal fibrosis. Given that stem cells are used as a promising therapeutic approach for fibrotic diseases, little effort has been undertaken to use stem cells to prevent post‐operative skeletal fibrosis. This review aims at providing useful information for the potential application of stem cells in preventing post‐operative skeletal fibrosis in the near future. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1236–1245, 2019.     

Transplantation of human CD34+ stem cells from umbilical cord blood to rats with thioacetamide-induced liver cirrhosis

BACKGROUND: Liver fibrosis results from accumulation of extracellular matrix components and is associated with many chronic hepatic diseases. There is to date no specific therapy for this disease, and patients receive treatment for its associated complications. Specific progenitor cells, known as oval cells, are present in the liver. As oval cells express markers such as CD34, they are thought to arise from a hematopoietic precursor. The aim of this work was to investigate whether transplantation of hematopoietic CD34(+) stem cells could improve hepatic fibrosis by their differentiation into hepatocytes. METHODS: CD34(+) stem cells from human umbilical cord blood were purified, transduced with a lentiviral vector containing the green fluorescent protein (GFP) gene and injected via portal vein into rats with liver cirrhosis induced by the 4-month administration of thioacetamide. Rats were killed 15 and 60 days post-transplantation. RESULTS: Up to 37% and 22% fluorescent cells were observed in the blood of control and cirrhotic rats, respectively, at 15 days post-transplantation. At 60 days post-transplantation, however, fluorescent cells were completely absent from the blood. Fluorescence was not detected in liver sections at either 15 or 60 days post-transplantation. Polymerase chain-reaction study to detect the GFP gene ruled out silencing of the transgene. CONCLUSIONS: These results suggest that the transplanted cells did not engraft in the liver and were eliminated from the rats.     

Combined effects of moderately elevated blood glucose and locally produced TGF-beta1 on glomerular morphology and renal collagen production.

BACKGROUND: There is a correlation between renal graft rejection and blood glucose (BG) levels. Furthermore, diabetic patients may develop non-diabetic renal diseases, which in some circumstances progress rapidly. Since transforming growth factor-beta1 (TGF-beta) levels are elevated in many renal diseases, the accelerated progression may be due to interactions between glucose and locally produced TGF-beta1. Therefore, we investigated the effect of mild hyperglycaemia on glomerular morphology and collagen production in TGF-beta1 transgenic mice. METHODS: To achieve BG concentrations of approximately 15 mmol/l in TGF-beta1 transgenic and non-transgenic mice, we used multiple streptozotocin (STZ) injections, and after 8 weeks, we measured the changes in glomerular morphology and total collagen content. We also analysed extracellular matrix (ECM) and protease mRNA levels using real-time polymerase chain reaction (PCR) and phosphorylated extracellular signal-regulated kinase 1/2 (pERK) expression by immunohistochemistry. RESULTS: Mild hyperglycaemia alone had no effect on glomerular structure or ECM deposition. Over-expression of TGF-beta1 increased basement membrane thickness (BMT) and the mesangial volume fraction. Furthermore, it augmented ECM, Matrix metalloproteinase-2 (MMP), MMP-9, plasminogen activator inhibitor-1 (PAI) and tissue inhibitor of metalloproteinase-1 (TIMP) gene expression and pERK1/2 immunostaining. Elevated BG in combination with TGF-beta1 resulted in enlargement of glomerular volume, total mesangial volume and renal collagen content. Moreover, high BG exaggerated TGF-beta1-induced changes in the BMT, MMP-2 and TIMP-1 expression and pERK1/2 staining. CONCLUSION: Even moderate elevations in BG accelerate the progression of those kidney diseases in which TGF-beta1 is involved. This emphasizes the importance of strict BG control in renal transplant patients and diabetic patients with renal malfunctions unrelated to diabetes.     

曲尼司特对环孢素A诱导的人肾小管上皮细胞向间充质转变的抑制作用

Objective To study the effect of tranilast on cyclosporine A (CsA)-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells, and investigate the mechanism of its antifibrotic effect. Methods Cultured HK-2 cells were divided into four groups: (1)In the control group, cells were treated without any medicine; (2) The cell were treated with CsA (4. 2μmol/L) for 72 h; (3) The cells were treated with a combination of CsA (4. 2 μmol/L) and tranilast (100μmol/L); (4) The cells were treated with tranilast (100 μmol/L) alone for 72 h.Morphological changes of the cells were assessed by phase-contrast microscopy. The immunofluorescence and Western blotting were adopted to detect the expression of E-cadherin, α-SMA and OPN mRNA and proteins respectively. Results Tranilast could markedly ameliorate the morphological changes of HK-2 cells stimulated by CsA. The irmmunofluorescence staining revealed the expression of E-cadherin was markedly decreased in HK-2 cells stimulated with CsA for 72 as compared with the control group, while the expression of α-SMA and OPN was significantly higher in CsA group than the control group. The expression of E-cadherin in the CsA + Tranilast group was higher than the CsA group, while the expression of α-SMA and OPN in the CsA + Tranilast group was lower than the CsA group. Western blotting showed that protein expression level of E-cadherin in CsA group was dramatically lower than that in the control group (P＜0. 05), while that of α-SMA and OPN in CsA group was significantly higher than in the control group (P＜0.05). The protein expression level of E-cadherin in HK-2 cells in the CsA + Tranilast group was markedly higher than in the CsA group (P＜0.05), and that of α-SMA and OPN in CsA + Tranilast group was significantly lower than in the CsA group (P＜0. 05). Conclusion Tranilast can block the CsA-induced epithelialto-mesenchymal transition in HK-2 cells probably by suppressing the expression of OPN.     

曲尼司特对环孢素A诱导的人肾小管上皮细胞向间充质转变的抑制作用

Objective To study the effect of tranilast on cyclosporine A (CsA)-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells, and investigate the mechanism of its antifibrotic effect. Methods Cultured HK-2 cells were divided into four groups: (1)In the control group, cells were treated without any medicine; (2) The cell were treated with CsA (4. 2μmol/L) for 72 h; (3) The cells were treated with a combination of CsA (4. 2 μmol/L) and tranilast (100μmol/L); (4) The cells were treated with tranilast (100 μmol/L) alone for 72 h.Morphological changes of the cells were assessed by phase-contrast microscopy. The immunofluorescence and Western blotting were adopted to detect the expression of E-cadherin, α-SMA and OPN mRNA and proteins respectively. Results Tranilast could markedly ameliorate the morphological changes of HK-2 cells stimulated by CsA. The irmmunofluorescence staining revealed the expression of E-cadherin was markedly decreased in HK-2 cells stimulated with CsA for 72 as compared with the control group, while the expression of α-SMA and OPN was significantly higher in CsA group than the control group. The expression of E-cadherin in the CsA + Tranilast group was higher than the CsA group, while the expression of α-SMA and OPN in the CsA + Tranilast group was lower than the CsA group. Western blotting showed that protein expression level of E-cadherin in CsA group was dramatically lower than that in the control group (P＜0. 05), while that of α-SMA and OPN in CsA group was significantly higher than in the control group (P＜0.05). The protein expression level of E-cadherin in HK-2 cells in the CsA + Tranilast group was markedly higher than in the CsA group (P＜0.05), and that of α-SMA and OPN in CsA + Tranilast group was significantly lower than in the CsA group (P＜0. 05). Conclusion Tranilast can block the CsA-induced epithelialto-mesenchymal transition in HK-2 cells probably by suppressing the expression of OPN.     

Quantifying fluctuation in glucose levels to identify early changes in glucose homeostasis in cystic fibrosis

Background

Cystic fibrosis related diabetes (CFRD) is associated with increased morbidity in CF. Variability in physiological systems is associated with dysfunctional homeostasis. We examined whether fluctuation in glucose is a marker of CFRD or “pre-diabetes”.

曲尼司特对环孢素A诱导的人肾小管上皮细胞向间充质转变的抑制作用

Objective To study the effect of tranilast on cyclosporine A (CsA)-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells, and investigate the mechanism of its antifibrotic effect. Methods Cultured HK-2 cells were divided into four groups: (1)In the control group, cells were treated without any medicine; (2) The cell were treated with CsA (4. 2μmol/L) for 72 h; (3) The cells were treated with a combination of CsA (4. 2 μmol/L) and tranilast (100μmol/L); (4) The cells were treated with tranilast (100 μmol/L) alone for 72 h.Morphological changes of the cells were assessed by phase-contrast microscopy. The immunofluorescence and Western blotting were adopted to detect the expression of E-cadherin, α-SMA and OPN mRNA and proteins respectively. Results Tranilast could markedly ameliorate the morphological changes of HK-2 cells stimulated by CsA. The irmmunofluorescence staining revealed the expression of E-cadherin was markedly decreased in HK-2 cells stimulated with CsA for 72 as compared with the control group, while the expression of α-SMA and OPN was significantly higher in CsA group than the control group. The expression of E-cadherin in the CsA + Tranilast group was higher than the CsA group, while the expression of α-SMA and OPN in the CsA + Tranilast group was lower than the CsA group. Western blotting showed that protein expression level of E-cadherin in CsA group was dramatically lower than that in the control group (P＜0. 05), while that of α-SMA and OPN in CsA group was significantly higher than in the control group (P＜0.05). The protein expression level of E-cadherin in HK-2 cells in the CsA + Tranilast group was markedly higher than in the CsA group (P＜0.05), and that of α-SMA and OPN in CsA + Tranilast group was significantly lower than in the CsA group (P＜0. 05). Conclusion Tranilast can block the CsA-induced epithelialto-mesenchymal transition in HK-2 cells probably by suppressing the expression of OPN.     

Tumor-Forming Idiopathic Retroperitoneal Fibrosis: Report of a Case

Idiopathic retroperitoneal fibrosis (IRF) is characterized by the progressive proliferation of connective tissue, but it rarely results in the formation of a mass. Herein, we report a rare case of tumor-forming IRF. A 76-year-old woman was referred to our hospital after a tumor in the right retroperitoneum was found by ultrasonography and computed tomography. Magnetic resonance imaging showed a 5 × 8 × 5-cm irregularly shaped tumor, lying adjacent to the right kidney, with a high-intensity T1-weighted image and a high-intensity T2-weighted image. Hormonal levels were within normal limits. Surgery was performed because of the possibility of an adrenal cancer. The tumor was firm, measured 7 × 8 × 4cm, and weighed 115g. The pathological diagnosis was retroperitoneal fibrosis. It is very difficult to distinguish tumor-forming IRF from malignancy. Several examinations, including needle aspiration cytology and biopsy, are necessary for the diagnosis and treatment of this disease.     

腹膜后纤维化致肾积水(附五例报告)

目的　提高腹膜后纤维化的诊治水平。　方法　报告收治的腹膜后纤维化致肾积水患者５ 例。　结果　原发性腹膜后纤维化４ 例,采用输尿管松解、带蒂大网膜包裹治疗,随访６ ～４０ 个月,肾功能正常,肾积水基本消失。继发性腹膜后纤维化１ 例,因肾积水严重,行患肾切除,随访３６ 个月,对侧肾功能正常。　结论　逆行肾盂造影是腹膜后纤维化重要的诊断手段,带蒂大网膜包裹术是有效的治疗方法。     

An international/multicentre report on patients with cystic fibrosis (CF) over the age of 40 years

BackgroundThe lifespan of patients with cystic fibrosis (CF) is increasing significantly. The objective of this international pilot study was to study the characteristics of these long-term survivors.MethodsFour centres with large CF clinics from London (UK), Minneapolis (USA), Toronto (Canada) and Verona (Italy) identified 366 patients who had survived 40years and longer.ResultsAt all centres males survived longer than females. There were more pancreatic sufficient patients in Verona (60%) and Toronto (40%) than in London (16%) and Minneapolis (21%). The percentage of ΔF508 homozygous patients varied between 47% in London and 45% in Minneapolis to only 26% in Toronto and 9% in Verona.Average FEV₁ and BMI values of the surviving population appeared to stabilise after 40years of age. FEV₁ was on average 12% higher in patients who were pancreatic sufficient (p > 0.0001). There was no difference in survival between the centres. The overall median survival after the age of 40 was 13years. The estimated annual death rate was approximately 3.4% from the age of 40–60years.ConclusionsSignificant numbers of patients are now surviving to 40years or more, and it is hoped that an in-depth study of these patients may identify the factors contributing to longer survival.     

Peritoneal fibrosis and its prevention

SUMMARY: Peritoneal fibrosing syndrome (PFS) is composed of a wide spectrum of peritoneal alterations observed in patients under peritoneal dialysis (PD). Long-term peritoneal exposure to unphysiological PD solutions and recurrent bacterial peritonitis had been claimed as the most common causes predisposing to the development of PFS in a PD population. With the advances in molecular research, physicians and pathologists recognized that peritoneal injury and the accompanied accumulation of extracellular matrix (ECM) within the peritoneum are key events leading to PFS. Bioincompatible solution and it's related products, inflammatory mediators, growth factors as well as cytokines in the peritoneal cavity are contributing factors. Therapeutic strategies antagonizing these mediators and/or their downstream intracellular signalling pathways with either drug molecules or gene transfer may have potential for the prevention or treatment of PFS.     

The management of anaesthesia for patients with cystic fibrosis

The clinical problems of patients with cystic fibrosis who require anaesthesia are reviewed. The management and the complications of anaesthesia in 77 patients over a 3-year period are presented. The long term effects of anaesthesia are analysed by a study of the lung function tests of these patients. The results suggest that even multiple procedures, as for instance nasal polypectomy, are not harmful. A method for safe peroperative management is presented.     

腹膜后纤维化行腹腔镜输尿管松解腹腔内间置术1例报道

目的 探讨腹膜后纤维化所致肾积水腹腔镜治疗的可行性.方法 报告1例因腹膜后纤维化所致左肾积水行腹腔镜治疗的诊治过程和结果,结合文献复习,探讨采用腹腔镜方法治疗腹膜后纤维化所致肾积水的可行性.55岁女性,间断左下肢浮肿,伴恶心、呕吐5个月,化验检查提示血肌酐和尿素氮进行性上升,血肌酐最高达503μmol/L.CT提示右肾萎缩,左输尿管下段与髂血管相交处狭窄,腹主动脉前与髂内外血管周围软组织影包绕,其上输尿管扩张.术前予以左输尿管双J管置入,手术取右侧半卧位,分别采用脐下1 cm、4 cm和左侧腹外斜肌外侧相同水平2点穿刺置入穿刺套管,腹腔镜自脐下1 cm处套管置人,术中见后腹膜呈板状,灰白色,打开后腹膜,见左输尿管巾段与髂血管周围粘连严重,超声刀仔细分离长度约9 cm,分离松解输尿管与周围组织问粘连,上下各达无粘连处,将此段输尿管放入腹腔内,关闭后腹膜.结果 术后恢复好,5 d切凵拆线,术后1月拔除双J管,拔管后3 h出现腰痛、发热和少尿,予以解痉、镇痛和抗菌治疗2 d后逐渐好转,尿量正常,多次复查肾功能正常,B超提示左肾积水程度逐渐减轻.3个月后复查B超提示左肾积水程度较术前显著减轻,肾功能维持正常.结论 腹膜后纤维化行腹腔镜输尿管松解、腹腔内间置术具有微创、病人痛苦小、术后恢复快的优点.但因有关此种治疗的报告例数少、随访时间较短,今后还需要更多病例和长时间的随访来进行验证其治疗效果.     

Presentation of idiopathic retroperitoneal fibrosis in the pediatric population

Idiopathic fibrosis of the retroperitoneum is rare in childhood. The authors describe an 11-year-old boy who presented with progressive renal failure, bilateral hydronephrosis, hypertension, and elevated erythrocyte sedimentation rate (ESR) owing to retroperitoneal fibrosis. Ureterolysis was performed with improvement in his creatinine level and blood pressure. The soft tissue mass consisted of dense collagenous fibers consistent with retroperitoneal fibrosis. Postoperatively, he received steroids and azathioprine. Retroperitoneal fibrosis in the pediatric population is rare with only 23 cases reported in the English-language literature. Treatment includes pulsed steroid regimens, ureteral catheterization, and retroperitoneal exploration with ureterolysis. If allowed to progress, renal failure can result and lead to death. The etiology of retroperitoneal fibrosis in the pediatric patient may include autoimmune diseases, infection, and neoplasm, but most cases are idiopathic. Retroperitoneal fibrosis should be considered in patients with an elevated ESR, hypertension, renal failure, and hydronephrosis. Evaluation also should include a search for autoimmune diseases and malignancy.     

Combined heart-lung-liver, double lung-liver, and isolated liver transplantation for cystic fibrosis in children

Abstract Between June 1990 and September 1995, 8 of 24 children with cystic fibrosis (CF) who were accepted either for combined transplantation or isolated liver transplantation died while waiting for a graft; 11 underwent transplantation and 5 are currently on the waiting list. Of the 11 children who had surgery, 7 (group 1) underwent one of the following procedures: heart-lung-liver ( n = 4), sequential double lung-liver ( n = 2), or bilateral lobar lung from a split left lung and reduced liver ( n = 1). During the same period, the four other children (group 2) underwent isolated liver transplantation (three full-size livers, one partial liver). There was one perioperative death in each group. Pulmonary infection was the most common cause of morbidity in group 1. Other complications in group 1 included tracheobronchial stenosis ( n = 2), biliary stricture ( n = 2), and severe ascites ( n = 2). All were successfully treated. Oblit-erative bronchiolitis developed in three patients. This was treated with FK 506. In group 2, pulmonary function tests improved or remained stable after liver transplantation. Surgical complications in group 2 included severe ascites ( n = 1), biliary stricture ( n = 1), and abscess of the liver ( n = 1). Actuarial survival was 85.7 %% 2 % in group 1 at 1 year; it remained unchanged at 3 years and was 64.2 % at 5 years.     

Association between glucose intolerance and bacterial colonisation in an adult population with cystic fibrosis,emergence of Stenotrophomonas maltophilia

Background

Diabetes is common in cystic fibrosis (CF). Glucose can be detected in the airway when the blood glucose is elevated, which favours bacterial growth. We investigated the relationship between dysglycemia and lung pathogens in CF.