A high-throughput three-dimensional (3D) hepatocyte culture model is constructed in this study. It is capable of replicating the 3D in vivo environment and offers the advantages of high throughput, enhanced reproducibility, low cost, and simplified operation, rendering a valuable tool for hepatotoxicity screening of traditional Chinese medicine (TCM). First, we constructed the 3D high throughput liver chip model using collagen hydrogel. The precision and its difference with traditional cell culture plates were assessed using acetaminophen, Tripterygium hypoglaucum, and Qili San as the references. The feasibility of this model was also investigated by comparing the hepatotoxicity among four batches of T. hypoglaucum and Qili San. The methodology verification shows that the 3D hepatocyte model is better than traditional two-dimensional (2D) cell culture model in precision and feasibility. Subsequently, we compared the hepatotoxicity of different samples over a period of three or seven days and found that the hepatotoxicity of TCM extracts increased over time. Finally, we compared the hepatotoxicity of T. hypoglaucum and its compound formula Kunxian capsule under equivalent concentration. The results showed that the compound Kunxian capsule could significantly reduce hepatotoxicity of T. hypoglaucum. Generally, we constructed a high-throughput, robust 3D liver-on-a-chip model with the potential of rapid assessing TCM-induced liver toxicity.

Purpose Estimation of the degree of pulmonary artery stenosis (PS) in children patients before treatment can provide an important basis for the choice of treatment.This study explores the accuracy of non-invasive continuous wave Doppler (CW) and electrocardiogram (ECG) in estimating the degree of PS in children patients.Materials and Methods Sixty consecutive cases of PS children were collected from January 2012 to August 2016 in the General Hospital of Shenyang Military.The right ventricular pressure was estimated by measuring cross-pulmonary artery pressure gradient by CW,or estimated by measuring the amplitude of the V1R wave by ECG,which was then compared with that measured by cardiac catheterization respectively.Results The right ventricular pressure estimated by measuring cross-pulmonary artery pressure gradient by CW was positively correlated with that measured by cardiac catheterization (r=0.88,P＜0.05).The right ventricular pressure estimated by measuring the amplitude of the V1R wave by ECG was also positively correlated with that measured by cardiac catheterization (r=0.83,P＜0.05).Conclusion The right ventricular pressure estimated by CW or ECG has good consistency with that measured by cardiac catheterization.Both CW and ECG can be used as noninvasive methods for estimating the degree of PS in children.

Forensic Medicine ; Humans ; Kidney Transplantation ; Polycystic Kidney Diseases

Objective To report our experience with retroperitoneoscopic nephroureterectomy with excision of a bladder-cuff for renal pelvic and ureteral tumors.Methods Thirty-five patients (21 women and 14 men;mean age,67 years;age range,49 -82 years) with upper urinary tract tumors underwent retro- peritoneal laparoscopic nephrourcterectomy with excision of a bladder-cuff.Of the 35 cases,15 had pelvic tumors and 20 had ureteral tumors;19 cases had the tumors on the right side and 16 on the left.Two cases had ureteral tumors combined with bladder tumors.One case had bilateral ureteral tumors then concomitantly had bladder tumors.The needle electrode was used to circleround incise the bladder thoroughly 0.5 cm away from the ureterostoma.Three trocars in the waist were used for dissecting the kidney;and the ureter was dis- sected as far distally downward.Then an incision of 5-9 cm was created in the lower abdomen to allow dis- section of the distal ureter and bladder-cuff and intact specimen extraction.Results The operation was successful in all 35 patients.The mean operative time was 3.1 h ( range,1.5-6.0h).The mean estimated blood loss was 166 ml (range,20-1600 ml).Four cases received blood transfusion.The patient's activity re- covered in 20-32 h after operation.Postoperative pathology showed transitional cell carcinoma in 30 cases, poorly differentiated adenocareinoma in 2 (ureter),squamous cell carcinoma in 1 (ureter),leiomyosarcoma in 1 (ureter),xanthogranulomatous pyelonephritis in 1.Duodenal leakage occurred in 1 patient who had had dialysis a drainage catheter was placed on the third d after operation,and the patient died of heart failure af- ter 2 months.Postoperative vesical irrigation was performed to prevent tumor recurrence.The mean hospital stay was 11 d.During a mean follow-up of 14 months(range,1-32 months),1 patient developed pelvic me- tastasis and was alive with the tumor.The other 33 patients survived free of tumor to date.No patient had re- current transitional cell carcinoma of the bladder.Conclusions Our data demonstrate that retroperitoneo- scopic nephroureterectomy for renal pelvic and ureteral tumors has shorter incision and more rapid postopera- tive recovery compared with open surgery.Using resectoscope to resect the termination of ureter allows more complete excision of the ureter.

Decoction is one of the most commonly used dosage forms of traditional Chinese medicine. The stability of chemical constituents in decoction is closely related to the clinical efficacy and safety. There were few reports about the influence of metal ions in the stability of chemical constituents in traditional Chinese medicine. However, there is no evidence that metal ions in decoction water need to be controlled. In this study, 2,3,5,4'-tetrahydroxy stilbene-2-O-β-D-glucoside (THSG), one of the main constituents in Polygoni Multiflori Radix was studied. Ordinary tap water, deionized water, and water containing different metal ions were used to investigate and compare the influence on THSG. The results showed that after storage in a dark place at the room temperature for 10 days, the degradation of THSG was 7% in deionized water, while undetectable in tap water. The content of THSG could be decreased by different kinds of metal ions, and the effect was concentration-dependent. Moreover, Fe3+ and Fe2+ showed the greatest influence at the same concentration; and our study has shown that THSG decreased more than 98% in Fe and Fe2+ solutions at 500 ppm concentration. In the same time we found out p-hydroxybenzaldehyde (molecular weight: 122.036 7) and 2,3,5-trihydroxybenzaldehyde-2-O-glycoside (molecular weight: 316.079 4) were the main degradation products of THSG in tap water and water containing Cu2+, Ca2+, Zn2+, Mg2+ and Al3+. The product of THSG dimer with a water molecule was found in water containing Fe3+ and Fe2+. The above results showed that the metal ions in water could significantly influence the stability of THSG in water, indicating that the clinical efficacy and safety of decoction would be affected if the metal ions in water were not under control. It's suggested that deionized water should be used in the preparation of decoction containing Polygoni Multiflori Radix in the clinic to avoid degradation of THSG. Meanwhile, decoction prepared by tap water should be taken by patients in a short time. Our investigation provides important information and reference about the influence of metal ions on the stability of decoctions in other traditional Chinese medicine that have unstable groups such as hydroxyls and unsaturated bonds, etc.
Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Ions ; chemistry ; Metals ; chemistry ; Plant Roots ; chemistry ; Polygonaceae ; chemistry ; Stilbenes ; chemistry

Currently, power generation in China is dominated by thermal power, wind power, nuclear power, and hydropower enterprises. The power source mainly comes from thermal power generation. The occupational hazards in thermal power station are noise, high temperature, power frequency electric fields, dust, and chemical toxins and so on, with noise and dust (silica and coal dust) being the primary factors. The occupational hazards in wind power station are noise, power frequency electric fields, high temperature, low temperature, and chemical toxins (sulfur hexafluoride, toluene, styrene, etc.), with noise and power frequency electric fields being the major concerns. The occupational radiation hazards in nuclear power station are gamma rays, beta rays, X-rays, neutrons, alpha rays, and radioactive aerosols. There is special attention in radiation protection but not enough protection in non-radioactive hazards such as noise, high temperature, and ammonia. The occupational hazards in hydropower station are noise, power frequency electric fields, vibration, radon and its de-composites, and chemical toxins, with noise and power frequency electric fields being the primary hazards. Different categories of power generation enterprises should identify key hazards and work site for occupational disease prevention and control based on the features of occupational hazards. Improving occupational health management and protection levels are essential measures.

BACKGROUNDFrom limited exposure with management of the native distal ureter ipsilateral to the transplanted kidney, we usually choose open nephroureterectomy (NU) or laparoscopic NU combined with an open approach in renal transplant (RTx) recipients. We herein describe our preliminary experience with total endoscopic NU with bladder cuff (BC) excision and evaluate its feasibility for RTx recipients.

METHODSFrom August 2008 to June 2011, eight RTx recipients underwent total endoscopic NU with BC excision for clinically presumed native upper urinary tract urothelial carcinoma (UUT-UC) ipsilateral to the transplanted kidney. Cystoscopic circumferential excision of the ipsilateral ureteral orifice with BC was followed by retroperitoneal laparoscopic NU using early ureteral ligation without primary BC closure. The intact specimen was removed through a 3-cm flank incision (an enlarged trocar site). Perioperative and pathological data and oncological outcomes were collected and analyzed.

RESULTSAll endoscopic procedures were completed successfully without major complications and with open conversion. The mean estimated blood loss was 100 ml with no blood transfusion. The mean operating room time was 234.8 minutes, mean time to ambulation was 2.6 days, and mean hospital stay was 9.0 days. Pathological findings confirmed UUT-UC in seven recipients, two with bladder UC. During the mean 25.2-month follow-up, none of the recipients developed recurrence, while two developed contralateral UUT-UC after the first NU.

CONCLUSIONTotal endoscopic NU with BC excision is technically feasible and safe for RTx recipients.

Adult ; Aged ; Aged, 80 and over ; Endoscopy ; methods ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Nephrectomy ; methods ; Retrospective Studies ; Ureter ; surgery ; Urinary Bladder ; surgery ; Urinary Bladder Neoplasms ; surgery

BACKGROUNDSimultaneous pancreas-kidney transplantation (SPKT) frees the diabetic patient with end-stage nephropathy from dialysis and daily insulin injections. Herein, we review consecutive cases of SPKT with bladder drainage performed at our institution over an 8-year period.

METHODSThe study population included 21 patients (16 males and 5 females) who underwent SPKT between September 2001 and September 2009. Seven patients had type-1 diabetes and 14 had type-2 diabetes. Nineteen patients were on dialysis at the time of transplantation. Donation after cardiac death donors were selected for SPKT. The mean human leukocyte antigen match was 2 (range 0 - 4). SPKT was always performed using bladder drainage and vascular anastomoses to the systemic circulation. Immunosuppressive treatment consisted of anti-lymphocyte globulin induction followed by tacrolimus, mycophenolate mofetil, and prednisone.

RESULTSThe mean hospital stay was 45.43 days. After a mean follow-up of 39.4 months, survival rates for patient, kidney, and pancreas were 76.2%, 76.2%, and 66.7% at 1 year; 76.2%, 59.3%, and 55.6% at 5 years; and 57.1%, 39.5%, and 41.7% at 8 years, respectively. Major complications included anastomotic leaks, reflux pancreatitis, and rejection. Six patients died from septic shock (n = 3), duodenal stump leak (1), cardiac arrest (1), or renal failure (1). Eight kidney grafts were lost due to acute rejection (n = 2), chronic rejection (3), and death with a functioning graft (3). Pancreatic graft failure (9) was caused by thrombosis (n = 1), rejection (2), duodenal stump leak (1), and death with a functioning graft (5).

CONCLUSIONSSPKT is a valid therapeutic option for uremic diabetics although few hospitals in China can undertake SPKT.

Adult ; Diabetes Mellitus, Type 1 ; surgery ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Graft Rejection ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Male ; Middle Aged ; Pancreas Transplantation ; adverse effects ; mortality ; statistics & numerical data ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome ; Urinary Catheterization

Objective Type 1 diabetes mice model was established to investigate the changes of key enzymes involved in testosterone synthesis in testes of early diabetic mice.Methods Tatolly 20 male C57 mice were randomly divided into two groups:control and diabetic groups,and the diabetes mice were ip administered with a single dose of 150 mg/kg Streptozotocin.Four weeks after confirmation of diabetic model,the serum and testis were collected for further study.The qRT-PCR method was used to measure the expression of LHR and steroidogenesis synthetase StAR,P450scc,3β-HSD6,P450c17a1,and 17β-HSD3 mRNA.ELISA assay was performed to measure the levels of testosterone and luteinizing hormone (LH) in serum,and the enzymatic activities of 3β-HSD1,1P450c17 and 17β-HSD3 in testis tissue.Results Compared to control group,the levels of testosterone and LH of diabetic group declined significantly (P ＜ 0.05) after four weeks.The mRNA levels of LHR,StAR,CYP11a1,Hsd3b6,CYP17a1 and Hsd17b3,and enzymatic activities of 3β-HSD6,P450c17 and 17β-HSD3 were also decreased significantly compared with control group (P ＜ 0.05,0.01 and 0.001).Conclusion Expression of key enzymes of testosterone synthesis in testis of early diabetic mice decreases significantly.

To study the effect of micro (mi)RNA on cellular proliferation induced by hepatitis B x protein, HBx, in human liver cells and to investigate the underlying molecular mechanism of this cancer-related effect. The human L02 hepatocyte cell line was stably transfected with HBx (L02/HBx) or an HBx mutant (L02/HBx-d382) that induces higher levels of cellular proliferation. The differential miRNA expression profiles were determined by microarray analysis and confirmed by real-time PCR. Two miRNAs, miR-338-3p and miR-551b, that were found to be significantly down-regulated in the L02/HBx-d382 cells were selected for further study and transfected individually into cells using the lipofectamine procedure. The cell survival rate was analyzed by MTT assay, and cell cycles were assessed by flow cytometry. Expressions of cyclinD1, cyclinG1, and E2F1 were assessed by real-time PCR and Western blotting. Compared with the microarray miRNA profile of L02/pcDNA3.0 cells, six miRNAs were up-regulated and five miRNAs were down-regulated in the L02/HBx-d382 cells, while four miRNAs were up-regulated and 12 were down-regulated in the L02/HBx cells. The microarray results were consistent with real-time PCR results. Transfection of miR-338-3p and miR-551b significantly inhibited the cell survival rates (P less than 0.001) and induced G0/G1 phase cycle arrest. According to MTT results: for L02/HBx-d382 cells, compared with lipofectamine or non-transfected (NC) controls, the t value of miR-338-3p was 10.402, 9.133 and the t value of miR-551b was 8.763, 7.403; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 9.105, 8.074 and the t value of miR-551b was 7.673, 7.52. According to flow cytometry results: for L02/HBx-d382 cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 12.173, 11.107 and the t value of miR-551b was 15.364, 13.377; for L02/HBx cells, compared with lipofectamine or NC controls, the t value of miR-338-3p was 15.416, 13.378, and the t value of miR-551b was 13.276, 13.109. The protein levels of cyclinD1, cyclinG1, and E2F1 were significantly reduced by both miR-338-3p and miR-551b ( P less than 0.001). For L02/HBx-d382 cells, compared with lipofectamine or NC controls: E2F1 had t = 11.132, 10.031 and 12.017, 10.973, respectively; cyclinD1 had t = 15.654, 15.013 and 15.447, 14.733, respectively; cyclinG1 had t = 8.017, 7.661 and 7.402, 7.417, respectively. For L02/HBx cells, compared with lipofectamine or NC controls: E2F1 had t = 14.244, 13.331 and 15.022, 14.468, respectively; cyclinD1 had t = 8.695, 8.137 and 7.877, 7.503, respectively; cyclinG1 had t = 7.73, 7.471 and 7.596, 7.41, respectively. In contrast, the mRNA levels for E2F1, cyclinD1, and cylcinG1 showed no significant differences between the miRNA transfected cells and controls. Wild-type HBx and the high proliferation-inducing mutant HBx can influence the miRNA expression profile of L02 cells. HBx down-regulates miR-338-3p and miR-551b in L02 cells, and the high proliferation-inducing mutant has a more robust effect. The mechanism of miR-338-3p- or miR-551b-mediated cell growth inhibition appears to be related to the direct modulation of cyclinD1, cyclinG1, and E2F1.
Blotting, Western ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Line ; Cell Proliferation ; Cyclins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Genes, Viral ; Hepatitis B virus ; genetics ; metabolism ; Hepatocytes ; metabolism ; pathology ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Mutation ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; Transfection