Objective To investigate the effect of Yiqi Jiedu Recipe(YQ)on the proliferation,migration,and invasion of nasopharyngeal carcinoma cells,and to explore its mechanisms of action on proliferation,migration,and invasion through the TGF-β1/SMAD3 signaling pathway.Methods(1)The 5-8F cells were divided into four groups:solvent control group,YQ 0.5 mg·mL-1 group,YQ 1.0 mg·mL-1 group,and 5-fluorouracil 2 μg·mL-1 group.Cell proliferation was monitored using real-time cell analysis(RTCA).Wound healing experiment was conducted to assess cell migration.After 24 hours of drug intervention,transwell assay was employed to measure cell invasion.The protein expression levels of β-catenin,E-cadherin,N-cadherin,TGF-β1,and SMAD3 in the cells were evaluated using the Western Blot method.(2)The 5-8F cells were divided into five groups:solvent control group,TGF-β1 10 ng·mL-1 group,TGF-β1 10 ng·mL-1+YQ 1.0 mg·mL-1 group,YQ 1.0 mg·mL-1 group,and LY3200882 10 μmol·L-1 group.Cell proliferation was monitored using RTCA.Wound healing experiment was conducted to assess cell migration.After 24 hours of drug intervention,transwell assay was employed to measure cell invasion.The protein expression levels of β-catenin,E-cadherin,N-cadherin,TGF-β1,and SMAD3 in the cells were evaluated using Western Blot.(3)The nude mice were randomly assigned into the model group,YQ group,and 5-fluorouracil group.Subcutaneous injection of 5-8F cell suspension was performed to establish the xenograft nude mouse model of nasopharyngeal carcinoma.After the tumors reached a certain size,the 5-fluorouracil group received intraperitoneal injection of 5-Fu once every 2 days,while the other groups were orally administered corresponding drugs once a day for three consecutive weeks.Tumor volume was measured every 3 days.Western Blot was conducted to assess the protein expression levels of β-catenin,E-cadherin,and N-cadherin in the tissues of each group.Results Compared with the solvent control group,the proliferation curves of 5-8F cells in the YQ(0.5 mg·mL-1,1.0 mg·mL-1)groups showed a decrease.The migration and invasion abilities of the cells were both reduced(P＜0.05,P＜0.01).Additionally,the expression of E-cadherin protein significantly increased(P＜0.01),while the protein expression of β-catenin,N-cadherin,TGF-β1,and SMAD3 all decreased(P＜0.05,P＜0.01).Compared with the model group,the transplanted tumor volume of nasopharyngeal carcinoma in the YQ group significantly decreased(P＜0.05).Furthermore,the protein expression of β-catenin and N-cadherin in the transplanted tissues of the YQ group was significantly downregulated(P＜0.05,P＜0.01),while the expression of E-cadherin protein was significantly upregulated(P＜0.01).After the addition of the activator and inhibitor of TGF-β1 signaling pathway,compared with the YQ 1.0 mg·mL-1 group,the TGF-β1 10 ng·mL-1+YQ 1.0 mg·mL-1 group showed a significant increase in the expression of TGF-β1,SMAD3,β-catenin,and N-cadherin proteins(P＜0.05,P＜0.01)and obvious enhancement of the abilities of cell proliferation,migration,and invasion(P＜0.01).Conclusion Yiqi Jiedu Recipe can inhibit the proliferation,migration,and invasion by regulating the TGF-β1/SMAD3 signaling pathway.

Objective To investigate the role of bone marrow mesenchymal stem cells (BMSCs) pretreated with erythropoietin (EPO) in the prevention of acute rejection after renal transplantation in rats. Methods BMSCs were divided into five groups: control group (without EPO), group A (pretreated with EPO at a final concentration of 10 IU/mL), group B (pretreated with EPO at a final concentration of 100 IU/mL), group C (pretreated with EPO at a final concentration of 500 IU/mL) and group D (pretreated with EPO at a final concentration of 1 000 IU/mL). In each group, the BMSCs were cultured for 24 h and 48 h. The proliferation rate of the BMSCs was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The BMSCs were divided into two groups: BMSC group (without EPO) and EPO-BMSC group (pretreated with EPO at a final concentration of 500 IU/mL). After 48 h culture, Western blot was adopted to measure the expression level of CXC chemokine receptor (CXCR) 4 protein in BMSCs. Wistar rats were used as the donors, and SD rats were utilized as the recipients to establish the rat models with acute rejection after renal transplantation. The recipient rats were randomly divided into four groups (n=6 in each group) including the control group (without any intervention), EPO group (injection of 1 mL of solution containing 500 IU EPO via tail vein immediately after surgery), BMSC group (injection of 1 mL of solution containing 1×106/mL BMSCs via tail vein immediately after surgery) and EPO-BMSC group (injection of 1 mL of solution containing 1×106/mL BMSCs cultured in vitro with 500 IU/mL EPO via tail vein). The level of serum creatinine (Scr) level was determined by Scr detection kit. Western blot was used to detect the expression levels of interferon (IFN)-γ and interleukin (IL)-4 proteins. Results After 24 h culture, the proliferation rate of BMSCs did not significantly differ among all groups (all P>0.05). After 48 h culture, the proliferation rate of BMSCs in group C (pretreated with EPO at a final concentration of 500 IU/mL) was significantly higher than that in the control group (P<0.05). Compared with the BMSC group, the expression level of CXCR4 protein on the surface of BMSCs was higher in the EPO-BMSC group (P<0.05). At 1 d after renal transplantation, the levels of Scr did not significantly differ among all groups (all P>0.05). At 5 d after operation, the levels of Scr in the EPO, BMSC and EPO-BMSC groups were significantly lower than that in the control group (all P<0.05). The level of Scr in the EPO-BMSC group was markedly lower than those in the EPO and BMSC groups (both P<0.05). At postoperative 1 d and 5 d, the expression levels of IL-4 protein in the kidney tissues did not significantly differ among all groups (all P>0.05). At 1 d after surgery, compared with control group, the expression levels of IFN-γ protein and IFN-γ/IL-4 ratio in the renal tissues in the EPO, BMSC and EPO-BMSC groups were significantly decreased to varying extents (all P<0.05), and similar results were obtained at 5 d after surgery (all P<0.05). The expression levels of IFN-γ protein and IFN-γ/IL-4 ratio in the EPO-BMSC group were significantly lower than those in the EPO group and BMSC group (both P<0.05). Conclusions BMSCs pretreated with EPO can prevent the incidence of acute rejection after renal transplantation and protect the renal graft function.

Objective To investigate the effect and underlying mechanisms of ghrelin in a rat model of renal fibrosis. Methods Male Sprague-Dawley rats were divided into 4 groups , including sham operation +saline or brain gut peptide treatment group , model + saline or brain gut peptide treatment group. Unilateral ureteral obstruction (UUO) was established by left ureteral ligation. 7 days and 14 days after operation, the rats were sacrificed , while the kidney tissue of obstruction side was harvested for pathlogical changes through Masson coloration. Expression of α-smooth muscle actin (α-SMA), transforming growth factor beta1 (TGF-β1) and phosphorylated Smad3 (p-Smad3) in renal tissues were analyzed through immunohistochemistry. Expression of α-SMA and TGF-β1 mRNA was detected by real-time-PCR. Apoptosis kidneys cells were marked with TUNEL. Results Ghrelin inhibited renal fibrosis by reducing the production of collagen , restraining extracellular matrix (ECM) deposition and decreasing the expression of α-SMA. Meanwhile, ghrelin inhibited the accumulation of myofibroblasts by blocking the transforming growth factor-β1/Smad3 (TGF-β1/Smad3) signaling pathway. Moreover, ghrelin could attenuate renal tubular cell apoptosis induced by UUO injury. Conclusion Ghrelin can reduce renal fibrosis and renal cell apoptosis induced by UUO , demonstrating that ghrelin is a potent antifibrotic agent that may have therapeutic potential for patients with obstructive nephropathy.

Objective To explore the clinical features and feasibility of renal transplantation for end-stage renal disease (ESRD) following hematopoietic stem cell transplantation (HSCT).Method A retrospective study was done in one case of renal transplantation for ERSD following HSCT.Clinical manifestations were summarized and prognosis was described.The 22-year-old male recipient had received HLA allele matched related bone marrow transplantation from his sister in 2001 and accepted renal transplantation 14 years after HSCT because of delayed renal dysfunction.Donor was a cardiac death patient,the preoperative Panel Reactive Antibody Testing was negative and there were 1.5 HLA antigen mismatches of 6 HLA-A,B,DR antigens of donor and recipient.The recipient received immunosuppressive therapy of tacrolimus + mycophenolate mofetil + steroid after renal transplantation.Result The patient's renal function remained stable and serum creatinine level was 65 μmol/L.The outcome of the patient was fairly good during the follow-up period of short-term.Conclusion Renal transplant is a feasible alternative for patients with ESRD following HSCT.If the transplanted kidney and abbr.hematopoietic stem cells are from different donors,irnmunosuppressive treatment is essential after renal transplantation.Long-term follow-up and adjustment of immunosuppression treatment are needed to prevent and treat postoperative complications.

OBJECTIVETo evaluate the clinical value of serum 1,3-beta-D-glucan (BG) and galactomannan (GM) detection for early diagnosis of invasive aspergillosis (IA) in patients after renal transplantation.

METHODSBlood samples collected from 69 renal transplant recipients were divided into diagnosis group, clinical diagnosis group, suspected diagnosis group, and non-infected group for detection of serum BG and GM.

RESULTSThe mean serum levels of BG in the diagnosis group, clinical diagnosis group, and suspected diagnosis group were significantly higher than that in non-infected group (P<0.05). The sensitivity, specificity, and positive and negative predictive values of BG was 69.49%, 70%, 93.18% and 35.71% for IA diagnosis, respectively. The serum levels of GM in the 3 diagnosis groups were also significantly higher than that in the non-infected group (P<0.05) with the sensitivity, specificity, and positive and negative predictive values of 84.75%, 90%, 96.15% and 52.63% for IA diagnosis, respectively.

CONCLUSIONIncreased serum BG and GM levels can serve as the evidence for early diagnosis of IA with a high diagnostic sensitivity and specificity in renal transplant recipients.

Aspergillosis ; diagnosis ; Early Diagnosis ; Humans ; Kidney Transplantation ; Mannans ; blood ; Sensitivity and Specificity ; beta-Glucans ; blood

BACKGROUND:The rate of tuberculosis infection was high in patients after renal transplantation. Clinical manifestation is not typical, which brings inconvenience to diagnose.
OBJECTIVE:To summarize the diagnosis and therapeutic methods of tuberculosis infection after al ograft renal transplantation.
METHODS:Relevant diagnosis and therapeutic method of 13 patients with tuberculosis infection after renal transplantation were retrospectively analyzed in the Department of Organ Transplantation, Zhujiang Hospital of Southern Medical University from January 2010 to October 2013.
RESULTS AND CONCLUSION:The onset time was 4-120 months after operation;62%(8/13) patients within 18 months after transplantation. Patients affected long-period fever, mainly low-grade fever. Four cases were identified according to the history, imaging data in combination with positive pathogenic diagnosis. Five cases were identified according to the history, imaging data combined with lung biopsy histopathology. The remaining four cases were identified according to the history, imaging data with experimental effective anti tuberculosis treatment. Early pulmonary symptom was not obvious. Chest CT was helpful in early diagnosis and differential diagnosis. Al patients fol owed early, law, ful , right amount, combined with principles of anti-tuberculosis treatment, and treatment usual y lasted for 6-10 months. They were given combined anti-tuberculosis infection drugs, adjustment of immunosuppressive agents and five-ester capsule for liver protection therapy. Thirteen patients were alive, no deaths. Two cases with early infection without timely treatment suffered from acute rejection, leading to loss of graft function and returned to hemodialysis. The others were cured and left hospital. Renal function was normal after 6-month fol ow-up (serum creatinine). Results indicated that after renal transplantation, patients with pulmonary tuberculosis should be early detected, early diagnosed and early treated. CT guided biopsy can be used as an effective and feasible means for diagnosis and identification of smear negative pulmonary tuberculosis after renal transplantation. Adjustment of immune scheme, anti-tuberculosis treatment and five-ester capsule significantly reduced calcineurin inhibitor dose, and lessened their adverse reactions.

Objective To explore the optimized method of venous anastomosis for right donor kidney transplantation in rats.Methods Sprague Dawley (SD) rats were used as donors and recipients for homologous rat kidney transplantation.Both bilateral kidneys were harvested from the donor rats (n =45).Ninety rats were used as recipients and divided into 4 groups according to randomly digital table:In groups AC (n =15 each),the right donor kidneys were transplanted into the left nephridial pit of recipients,and endto-side,venous bypass and modified end-toend (donor's proximal end of vena cava was anastomosed to recipients renal Vein followed by ligation of its distal end) venous anastomosis was done,respectively; In the control group (n =45),the left donor kidneys were transplanted into the same side of the recipients,and the conventional end-to-end venous anastomosis was used.Then the intra-operative findings,successful operation rate and postoperative complications were compared between two groups.Results The venous anastomosis time in group B was longer than in groups A,C and control group (P＜0.05),which significantly increased warm ischemia time of donor kidneys and operative time of recipients (P＜0.05).The venous anastomosis time,warm ischemia time of donor kidneys and operative time of recipients showed no significant difference between groups A or C and control group (P＞0.05).The successful operation rate in group C (93.3％)was similar to that in control group (86.7％) (P＞0.05),but higher than in group A (53.3％) and group B (53.3％) (P＜0.05).There was no significant difference in postoperative complications between group A and group C.Conclusion For right donor kidney transplantation,the method of harvesting the right donor kidney with a part of vena cava,and then anastomosing the proximal end to recipients renal vein and ligating the distal end,is highly feasible,efficient and economic.

Objective Toobservetheindication, safetyandefficacyofanew immunosuppressant Rituximab in kidney transplantation. MethodsFive patients, who were diagnosed as antibody mediated rejection (AMR) from December 2010 to June 2011, were treated with single dose of Rituximab (500 mg) and followed up for 6 months. The clinical data, such as age, gender, onset of illness, induction therapy, maintaining therapy, allograft function, change of PRA, opportunistic infection and other complications were collected and retrospectively analyzed to evaluate the safety and efficacy of Rituximab used in AMR patients. ResultsAfter Rituximab therapy, all the patients had improved renal function measured by sera creatinine level: 4 cases retumed to normal, and 1 keep stable. Series of allograft biopsy demonstrated obviously reduced C4d deposition in nephridial tissue after treatment. One patient developed CMV viremia, another had urinary infection, but no one had lifethreatening infection during the follow-up period. The survival rate of human and allograft was both 100 ％. Conclusion Rituximab has a good efficacy and safety in treatment of AMR after renal transplantation.

Objective To evaluate the efficacy of percutaneous transluminal angioplasty (PTA) in the treatment of transplanted renal artery stenosis (TRAS)-induced renal dysfunction and hypertension. Methods Between July 1998 and January 2007, PTA was performed on 16 patients with RTAS. Color Doppler uhrasonography preceded the intra-arterial angiographic investigation,with false-negative results in 18. 75 % of patients. Sixteen cases of TRAS were examined at 1 st week,6th month and 13 years after PTA. Hypertension improvement was defined as mean arterial pressure decrease of at least 15 % from the pre-PTA value. Graft function was evaluated by SCr levels, and the improvement was defined as a 20% change. Results Angioplasty was technically feasible in 100 %.Sixteen patients with RTAS were cured clinically. During the follow-up period, graft function was improved in 81.25 %, 68. 75 %, 62. 5 %, 56. 25 %, 50 % of patients respectively at 1st week, 6th month and 1-3 years after PTA. The blood pressure was decreased in 62. 5%, 75 %, 75 %,56. 25 %, 50 % of patients respectively, but no patient remained hypotensor medication free.Conclusion PTA improved renal dysfunction and hypertension induced by TRAS, and it is a safe and effective treatment for TRAS.

BACKGROUND: Panel reactive antibody (PRA) can mediate hyperacute rejection, and lead to decrease in success rate of transplantation and survival rate of renal graft in highly sensitized recipients compared to non-sensitized recipients.OBJECTIVE: According to human leucocyte antigen (HLA) cross-matching standards to select suitable donors for sensitized recipients and to evaluate the incidence of acute rejection and survival rate of renal allografts.DESIGN: Case observation.SETTING: Zhujiang Hospital of Southern Medical University.PARTICIPANTS: 136 sensitized recipients with positive PRA underwent renal transplantation in Department of Organ Transplantation, Zhujiang Hospital of Southern Medical University between January 1997 and December 2003 were selected, including 41 males and 95 females, aged (45±9) years. Recipients of first, second, third, and fourth transplant were 115, 18, 2 and 1 case, respectively. The informed consent was obtained from all patients. The protocol was approved by Hospital Ethics Committee. Lambda antigen tray (LAT) and LAT-Mix were purchased from One Lambda, Inc, USA. Special monoclonal tray -Asian HLA class Ⅰ (SMT72R) and Micro SSP Generic HLA Class Ⅱ (DRB/DQB) were also purchased from One Lambda, Inc, USA.METHODS: Pre-operative PRA levels and specificity of recipients were detected by ELISA test with Lambda antigen tray (LAT). Donor and recipient HLA class Ⅰ typing was performed with special tray - Asian HLA class Ⅰ (SMT72R), and HLA class Ⅱ gene typing with Micro SSP Generic HLA Class Ⅱ (DRB/DQB) (Micro-SSP). HLA-matching between donor and recipient was performed according to HLA cross-reactive group (CREG) standards by UNOS and class Ⅱ antigen permissible mismatch. The incidence of acute rejection and survival rate of renal allografts were evaluated within 1, 3 and 5 years.MAIN OUTCOME MEASURES: ①PRA levels and specificity of sensitized recipients before and after transplantation; ②HLA-matching between donor and recipient; ③Incidence of acute rejection and survival rate of renal allografts after transplantation.RESULTS: 136 PRA positive sensitized recipients were all included in final analysis. ① There were 104 recipients with anti-HLA class Ⅰ IgG antibody, 76 with anti-HLA class Ⅱ IgG antibody, and 44 with both anti-HLA class Ⅰ and Ⅱ IgG antibodies in 136 recipients. ②The number of cases of 0, 1, 2, 3, and 4 mismatch (MM) was 7, 26, 47, 39 and 17, respectively by the standard of conventional HLA antigen matching; However, the number of the recipients with 0, 1, 2, 3, and 4MM was 31, 53, 36, 16, and 0, respectively according to the principle of HLA CREG matching. ③By the principle of HLA CREG matching, rates of acute rejection in sensitized recipients with 2MM and 3MM HLA-CREG were significantly higher than those with 0MM (P < 0.05). Renal allograft survival rate in sensitized recipients with 0MM was significantly higher than those with 2MM and 3MM (P < 0.05).CONCLUSION: ①HLA CREG matching can significantly improve the ratio of well-matched. ② Good HLA matching can reduce the incidence of acute rejection in sensitized recipients and increase the survival rate of renal grafts.