Biological upcycling of nickel and sulfate as electrocatalyst from electroplating wastewater.

Upcycling nickel (Ni) to useful catalyst is an appealing route to realize low-carbon treatment of electroplating wastewater and simultaneously recovering Ni resource, but has been restricted by the needs for costly membranes or consumption of large amount of chemicals in the existing upcycling processes. Herein, a biological upcycling route for synchronous recovery of Ni and sulfate as electrocatalysts, with certain amount of ferric salt (Fe3+) added to tune the product composition, is proposed. Efficient biosynthesis of bio-NiFeS nanoparticles from electroplating wastewater was achieved by harnessing the sulfate reduction and metal detoxification ability of Desulfovibrio vulgaris. The optimal bio-NiFeS, after further annealing at 300 °C, served as an efficient oxygen evolution electrocatalyst, achieving a current density of 10 mA·cm-1 at an overpotential of 247 mV and a Tafel slope of 60.2 mV·dec-1. It exhibited comparable electrocatalytic activity with the chemically-synthesized counterparts and outperformed the commercial RuO2. The feasibility of the biological upcycling approach for treating real Ni-containing electroplating wastewater was also demonstrated, achieving 99.5 % Ni2+removal and 41.0 % SO42- removal and enabling low-cost fabrication of electrocatalyst. Our work paves a new path for sustainable treatment of Ni-containing wastewater and may inspire technology innovations in recycling/ removal of various metal ions.

Microbiome profiling of nasal extracellular vesicles in patients with allergic rhinitis.

Background: Nasal microbiota is crucial for the pathogenesis of allergic rhinitis (AR), which has been reported to be different from that of healthy individuals. However, no study has investigated the microbiota in nasal extracellular vesicles (EVs). We aimed to compare the microbiome composition and diversity in EVs between AR patients and healthy controls (HCs) and reveal the potential metabolic mechanisms in AR. Methods: Eosinophil counts and serum immunoglobulin E (IgE) levels were measured in patients with AR (n = 20) and HCs (n = 19). Nasal EVs were identified using transmission electron microscopy and flow cytometry. 16S rRNA sequencing was used to profile the microbial communities. Alpha and beta diversities were analyzed to determine microbial diversity. Taxonomic abundance was analyzed based on the linear discriminant analysis effect size (LEfSe). Microbial metabolic pathways were characterized using Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUst2) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Results: Eosinophils, total serum IgE, and IgE specific to Dermatophagoides were increased in patients with AR. Alpha diversity in nasal EVs from patients with AR was lower than that in HCs. Beta diversity showed microbiome differences between the AR and HCs groups. The microbial abundance was distinct between AR and HCs at different taxonomic levels. Significantly higher levels of the genera Acetobacter, Mycoplasma, Escherichia, and Halomonas were observed in AR patients than in HCs. Conversely, Zoogloea, Streptococcus, Burkholderia, and Pseudomonas were more abundant in the HCs group than in the AR group. Moreover, 35 microbial metabolic pathways recognized in AR patients and HCs, and 25 pathways were more abundant in the AR group. Conclusion: Patients with AR had distinct microbiota characteristics in nasal EVs compared to that in HCs. The metabolic mechanisms of the microbiota that regulate AR development were also different. These findings show that nasal fluid may reflect the specific pattern of microbiome EVs in patients with AR.

[The preparation of StAR gene probe and its expression in mice Leydig cells under stress].

OBJECTIVE: To prepare the cDNA probe of the steroidogenic acute regulatory (StAR) gene, and to investigate the StAR mRNA expression in stressed C57BL/6 mice leydig cells. METHODS: cDNA fragment encoding mouse StAR was amplified by RT-PCR from the total RNA prepared from the testis, and then the RT-PCR product was cloned into pCR2. 1-TOPO vector. After StAR gene cDNA was sequenced, the Dig-labeled cRNA probes for mouse StAR gene were prepared by in vitro transcription from cDNA fragment. With the specific cRNA probes, in situ hybridization analysis was conducted in stressed mice testis and controls. RESULTS: The results demonstrated that StAR mRNA levels were significantly lower in stressed leydig cells than that in controls (P < 0.05). CONCLUSIONS: The decreased StAR mRNA levels induced by stress may result in a reduced production of StAR protein, which compromised the transportation of cholesterol, the substrate for androgen biosynthesis, into mitochondria, resulting in a poor T production in leydig cells and finally a declined T levels.

Laparoscopic treatment of a calcium fluorophosphate stone within a seminal vesicle cyst.

Stones in the seminal vesicles are extremely rare. We present a 62-year-old patient with a stone within a seminal vesicle cyst, who was cured by laparoscopic treatment. The operative time was 80 min, and the estimated blood loss was 90 mL. Scanning electron microscope examination of the stone showed a compact crystal image externally and sparse spherical crystal structure in kernel. Composition of the stone was calcium fluorophosphate on X-ray diffractometer. The follow-up time was 15 months with no recurrence of cyst or stone. To our knowledge, this case is the first to describe laparoscopic removal of a stone within a seminal vesicle cyst, and the first to describe calcium fluorophosphate as the composition of seminal vesicle stones.

[Effects of experimental varicocele on the testosterone level in the serum and testis of rats].

OBJECTIVE: To study the effects of experimental varicocele (VC) on the serum testosterone (T) and intratesticular testosterone in adolescent rats. METHODS: A VC rat model was established by partial ligation of the left kidney vein in 20 SD rats, and another 20 were included in a sham operation group as controls. At 4 and 8 weeks, the concentrations of the serum T and intratesticular T were measured by radioimmunoassay (RIA). The testis tissues were homogenized and the extract liquid taken for RIA. RESULTS: Compared with the controls, the level of serum T declined at 4 and 8 weeks in the VC group, but not significantly (P > 0.05), so was that of bilateral intratesticular T at 4 weeks, and with statistical significance at 8 (P < 0.01). CONCLUSION: Within 8 weeks, experimental VC could reduce the level of bilateral intratesticular T, but not that of serum T. Varicocele could damage Leydig cells.

[Expression of kinase insert domain-containing receptor in prostate adenocarcinoma].

OBJECTIVE: To investigate the correlation between the expression of the kinase insert domain-containing receptor (KDR) and the histological grade of prostate adenocarcinoma. METHODS: Forty-eight samples of prostate adenocarcinoma tissues and 20 samples of benign prostatic hypertrophy (BPH) tissues were studied by LsAB immunohistochemical staining. RESULTS: The positive expression rate of KDR was 73% in prostate adenocarcinoma and 30% in BPH. The expression of KDR was stronger in prostate adenocarcinoma, and there was no relationship between staining intensity and the histological grade of carcinoma. CONCLUSION: KDR, expressed more highly in prostate adenocarcinoma, promises to be a new target in the treatment of prostate adenocarcinoma.

[Biocompatibility of bladder extracellular matrix as tissue engineering scaffold].

OBJECTIVE: To investve To investigate the biocompatibility of rabbit bladder extracellular matrix (BECM) and evaluate the feasibility of using BECM as scaffold for reconstruction of tissue engineering urinary tract. METHODS: By the application of associating the solution diosmosis, enzymatic digestion with chemical detergent, the rabbit bladder was decellularized to prepare the BECM, on which rabbit bladder transitional epithelial cells were cultured and seeded in vitro, of which the adhesion and proliferation were observed. MTT method was used to evaluate the cytotoxicity of BECM, which was implanted into the rabbit back for evaluating its histocompatibility meant that the toxicity, degradation and local inflammatory response were studied through gross observation and HE staining. RESULTS: The prepared rabbit BECMs were freeze-drying, and looked like thin semitransparent membrane. By electron microscope examination, there were no residues of cells found on BECM membrane, of which one side had the reticular fibrous structure, and the other side had the compact structure. The co-culture of BECM membrane with bladder transitional epithelial cells indicated that BECM had a good cytocompatibility. The cytotoxicity score tested by MTT method was number 0 and 1. The rabbits in the implant test had no abnormal response. The BECMs degraded gradually in vivo, and growth of peripheral tissue could be seen in the materials after eight weeks. CONCLUSION: The BECMs prepared by our process are cell-free under scanning electron microscopy. Meanwhile, the reticular structure of the tissue matrix is well preserved. The BECMs have the good cytocompatibility with transitional epithelial cells but without cytotoxicity. The BECMs can degrade gradually in vivo with good histocompatibility. BECM is a good bio-derived material of tissue engineering as scaffold for urinary tract reconstruction.

[Explorative study on mechanism and drug intervention for chronic allograft nephropathy induced by renal ischemia-reperfusion injury].

OBJECTIVE: This study is aiming to investigate the mechanism and drug intervention of chronic allograft nephropathy (CAN) induced by renal ischemia-reperfusion injury. METHODS: The closed colony strain Sprague-Dawley (SD) and Wistar Rats were used as donor and recipient, respectively. Orthotopic kidney transplantation was performed following the procedure of our previous study. The rats were divided into five groups: Group A only received CsA with 10 mg/(kg x d); except CsA, Group B,C,D received Yi Sheng injection with 4 mg/(kg x d), 8 mg/(kg x d), and MMF (20 mg/(kg x d)], respectively. Group E was control group. According to Banff standard, the serum creatinine (SCr) level and pathological change of rat grafted kidney were observed at the 4th, 8th and 12th weeks post-transplantation. The immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction were used to comprehend the localization and expression of TGFbeta1 and Smad2, 7 in the transplant kidney. RESULTS: Compared with the lower dosage group, the differences of SCr level and pathological changes of CAN at all the time points after 8th week were statistically significant in the high dosage Yi Sheng group. It was showed that the Yi Sheng injection had the protective effect on CAN with a dose-dependent fashion. After transplantation, the rat kidney-sclerosis model showed that the up-regulated expressions of TGF-P, and Smad2 and the down-regulated expression of Smad7 in Group A and Group B were statistically significant, which meant that the difference was obvious when Group A compared with the other 4 groups. The expressions of TGF-beta1, and Smad2 were strongly positive in tubular epithelial cell, interstitial cell and glomerulus, while the expression of Smad7 was weak. Thickening of endovascular could significantly be inhibited in high dosage of Yi Sheng and MMF group. CONCLUSION: The up-regulated expressions of TGF-beta1 and Smad2 and the down-regulated expression of Smad7 may accelerate the progression of CAN alone or with immune factors. The traditional Chinese medicine Yi Sheng injection and MMF can down-regulate the expression of TGF-beta1 and Smad2 and block the down-regulated expression of Smad7, which may postpone and lessen the progression of CAN.

[Construction of tissue-engineered urothelial tissue in vitro and in vivo with bladder acellular matrix as scaffold].

OBJECTIVE: To investigate the construction of urothelial structure by tissue engineering. METHODS: Fresh bladder of New Zealand white rabbits were processed to prepare the bladder acellular matrix graft (BAMG) as the scaffold, which was evaluated by Masson's trichrome staining and the scanning electronic microscope. Bladder epithelia were obtained by enzymatic digestion and were proliferated in vitro. Growth of cells was observed under the inverted phase contrast microscope, and cells were identified by immunohistochemical method. The bladder epithelia were seeded on BAMG, and the epithelia/BAMG composites were observed by HE staining and the scanning electronic microscope. The composites fabricated in vitro were implanted into nude rats, and were retrieved in 4 and 8 weeks, which were observed by general observation, histological and immunohistochemical method. RESULTS: White semi-transparent membrane appeared in the prepared BAMG, and a fibre mesh structure of the material without residual cells was observed under the scanning electronic microscope. Bladder epithelia cultured in vitro showed a paving stone structure. Immunohistochemical staining with cytokeratin was performed, and brown cellular plasma staining was observed as positive reaction. After the epithelia were seeded on BAMG in vitro for 7 days, the cells fully covered the surface of the framework, showing a single-layer cellular structure. After being implanted into nude rats for 4 weeks and 8 weeks, the epithelia seeded on the BAMG formed a multi-layer structure. CONCLUSION: Urothelial structures can be constructed in vitro and in vivo by tissue engineering, which lays a technical foundation for further tissue engineered urinary tract reconstruction experiments.

Risk factors for prostatic inflammation extent and infection in benign prostatic hyperplasia.

AIM: To investigate the risk factors for prostatic inflammation extent and infection in patients with benign prostatic hyperplasia (BPH) so as to manage prostatic inflammation more efficiently. METHODS: Sixty patients with BPH undergoing TURP between September 2005 and December 2005 in West China Hospital of Sichuan University were studied. Prostate fluid (PF) was collected for the measurement of secretory IgA (SIgA) and complement 3 (C3). Prostate tissue were collected for testing bacterial 16S rDNA by real-time PCR, examining SIgA in the tissue and examining the inflammation. The possible clinical and immune risk factors for prostatic inflammation or infection were analyzed by using the logistic regression method. RESULTS: Abnormal white blood cell count in urinalysis, prostatic infection and a high concentration of C3 in PF are the risk factors for prostatic inflammation extent (P = 0.025, 0.034 and 0.035, respectively and odds ratio [OR] = 18.269, 8.284 and 1.508, respectively). Risk factors for prostatic infection include the C3 concentration and the concentration of SIgA in PF (P = 0.003 and 0.013, respectively, and OR=1.645 and 0.993, respectively). CONCLUSION: The present study suggests that prostatic inflammation is associated with urinary tract infection, prostatic infection and the activated complement and that prostatic infection is associated with the activated complement and downregulated mucosal immunity in prostates of the patients with BPH. It is also suggested that individual immune regulation should be considered in the treatment of prostatic inflammation and infection of patients with BPH.

[Apoptosis in male germ cells induced by testicular denervation].

OBJECTIVE: To evaluate the role of spermatic nerves in the regulation of spermatogenesis. METHODS: Fifty-four mature SD male rats (350-375 g) were randomized into a sham operation group (SO) and three experiment groups, and the latter underwent bilateral surgical removal of the superior spermatic nerve (SSN) or/and the inferior spermatic nerve (ISN). The animals were killed 1 month and 2 months after the operation. HE stain was used to observe spermatogenesis. Transmission electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) were employed to detect apoptosis. RESULTS: Impaired spermatogenesis was observed 2 months after the operation, with only Sertoli cells and a few spermatogonia remaining in the regressed tubules in all the treatment groups. The abnormal tubules in the SSN, ISN and SSN + ISN denervated testes accounted for (13.25 +/- 2.03)%, (11.0 +/- 4.36)% and (34.17 +/- 3.78)% respectively. Chromosome condensation and fragmentation in the germ cells were observed under the electron transmission microscope in all the denervated testes. TUNEL showed the spermatogonia and Leydig cells to be apoptotic in all the denervated testes and the incidence of the apoptotic cells in the SSN + ISN denervated testes was significantly higher than in the SSN or ISN denervated ones. CONCLUSION: Spermatic nerves play an important role in spermatogenesis.

[Expression of Bax and Bcl-2 gene in rat testis tissues induced by testicular denervation].

OBJECTIVE: To detect the changes of the expression of Bax and Bcl-2 gene in the denervated testis, and to explore the possible mechanisms underlying the apoptosis of germ cells induced by testicular denervation at the genetic translation level. METHODS: Eighteen mature SD rats (350-375 g) were equally divided into 3 groups: a sham operation group( SO) , a superior spermatic nerve group (SSN) and an inferior spermatic nerve group (ISN) , and the latter two received bilateral surgical removal of the superior spermatic nerve and the inferior spermatic nerve, respectively. The animals were killed I month after the operation. ISH SP-method was used to detect the expression of Bax and Bcl-2 protein. RESULTS: Significant up-regulation of Bax protein was detected in both the treatment groups 1 month after surgery( P <0. 05) , and the level of Bcl-2 protein remained unchanged. CONCLUSION: Bax gene is involved in the apoptosis of germ cells induced by testicular denervation.

[Effect of KLF6 on prostate cancer cell line PC-3 by transgenic method].

OBJECTIVE: To observe the effect of KLF6 on prostate cancer cell line PC-3 by transgenic method. METHODS: We obtained KLF6 cDNA by RT-PCR method from the liver cell, transfected plasmid pEGFP-C, recombinated with KLF6 into PC-3 cells, and used them as a transfection group and a control group. MTT, flow cytometer and immunocytochemical methods were used to observe the effect of anti-oncogene wild type KLF6 on prostate cancer cell line PC-3 by transgenic method for 48 hours. RESULTS: After transfected into PC-3 cells, KLF6 enhanced growth suppression, (30.0 +/- 5.4)% in the transfection group and 0% in the control, P < 0.01, apoptosis, (24.3 +/- 2.3)% in the transfection group and (5.2 +/- 0.7)% in the control, P < 0.01, the down-regulation of the expression of cyclin D1, (25.3 +/- 3.7)% in the transfection group and (38.5 +/- 4.6)% in the control, P < 0.05 and Bcl-2, (18.7 +/- 3.2)% in the transfection group, and (41.8 +/- 5.9)% in the control, P < 0.01 in PC-3 cells. It also decreased the ratio of the cell phase G2/M, increased the ratio of G0/G1 from (58.6 +/- 7.3)% in the control to (80.0 +/- 9.8)% in the transfection group, P < 0.05. CONCLUSION: PC-3 cells transfected with wild type KLF6 can enhance its growth suppression and apoptosis. It shows great potential for the gene therapy of androgen-independent carcinoma of the prostate.

[Effects of heating on the epithelia of benign prostate hyperplasia in vitro].

OBJECTIVE: To investigate the biological changes of the epithelia of benign prostate hyperplasia under heating at different temperatures and at different points of time after heating. METHODS: Cell morphology, MTT, flow cytometer, and the immunocytochemical method for detecting heat shock protein 70 and prostate specific antigen were used to observe the effect of heating on the primarily cultured epithelia of benign prostate hyperplasia at 45 degrees C and 60 degrees C and at 1 hour and 12 hours after heating. RESULTS: Heating at 45 degrees C for 15 min resulted in apoptosis, and at 60 degrees C, necrosis in most of the cells. The inhibiting effect of heating on the growth of cells was observed, more significant in the 60 degrees C group than in the 45 degrees C group. The cell phase arrest induced by heat, mainly G0/G1 arrest, was more significant in the 12 h group than in the 1 h group. Heating up-regulated the expressions of heat shock protein 70 and prostate specific antigen in cells. CONCLUSION: Heating can induce apoptosis, necrosis and growth suppression of the epithelia of benign prostate hyperplasia. Its process and mechanism are correlated with the cell phase arrest and the up-regulation of the expressions of heat shock protein 70 and prostate specific antigen.

[Determination of expression of protamine-2 mRNA in different positions of the testis of patients with nonobstructive azoospermia].

OBJECTIVE: To investigate the relationship between the expression of protamine-2 (P-2) mRNA and the results of sperm extraction in the corresponding testis tissues of patients with nonobstructive azoospermia. METHODS: Based on pathological diagnosis, 38 cases of azoospermia at the mean age of 32.4 (ranging 24 - 42) years were divided into a nonobstructive (NOA) group and an obstructive (OA) group. Two specimens were taken from different positions of one testis, each divided into three portions for general pathological test, sperm separation and mRNA extraction, respectively. The expression of P-2 mRNA was determined by RT-PCR and image analysis assay. RESULTS: Among the 38 cases, 27 were diagnosed as nonobstructive and 11 as obstructive azoospermia. No regularity was found as to the positions where sperm could or could not be successfully isolated. The expression of P-2 mRNA was 1.40 +/- 0.21 in the tissues where sperm was isolated and 0.51 +/- 0.23 (P < 0.05) in those where no sperm was isolated. CONCLUSION: The expression of P-2 mRNA in the testicular tissues from the patient with nonobstructive azoospermia could reveal the results of sperm extraction in the corresponding tissues.

[Constructing recombinant plasmid with prostate-specific membrane antigen promoter and reverse enhancer and comparing the transcription activity of enhancers in different directions].

OBJECTIVE: To assess the specificity of prostate-specific membrane antigen(PSMA) promoter and enhancer in controlling gene expression and to compare the activity of enhancers in different directions for choosing the most suitable prostate-specific PSMA controlling elements. METHODS: PSMA enhancer gene was amplified with PCR, then the enhancer gene was subcloned into the expressing vector pEGFP-PSMA(Pro) reversely to construct the recombinant plasmid pEGFP-PSMA(E(r)-p), which was transfected into different cell lines such as LNCaP, PC-3,MCF-7,A549. Green fluorescent protein (GFP) expression was observed and compared to other recombinants constructed previously. RESULTS: The recombinant plasmid with reverse enhancer was successfully constructed, the PSMA promoter and enhancer showed modulating activity in PSMA-expressed cell line uniquely. PSMA enhancer could increase 30-fold transcriptional activity over the basal level achieved by PSMA promoter alone, and no impact of the direction on the activity of enhancer was noted. CONCLUSION: PSMA promoter/ enhancer is specific to PSMA-expressed cells. The transcriptional activity of reverse enhancer is similar to that of enhancer. PSMA promoter/enhancer has the potential for use in targeted gene therapy of prostate adenocarcinoma.

[The construction of recombinant plasmid with prostate-specific membrane antigen promoter controlling reporter gene expression].

OBJECTIVE: To study the specificity of prostate-specific membrane antigen (PSMA) promoter in controlling gene expression. METHODS: PSMA promoter gene was amplified with PCR, and then the promoter gene was cloned into the vector pEGFP-1 to construct a recombinant plasmid, which was transfected into different cell lines such as LNcap, PC-3, MCF-7, A549. Green fluorescent protein (GFP) expression was observed. RESULTS: The recombinant plasmid constructed, and PSMA promoter uniquely showed modulating activity in PSMA positive cell line. CONCLUSION: PSMA promoter possesses PSMA positive cell specificity, as well as prostatic tissue specificity. PSMA promoter may have the potential for use in targeted gene therapy of prostate adenocarcinoma.

[Synergistic effects of paclitaxel and gemcitabine on androgen-independent prostate cancer].

OBJECTIVE: To observe the synergistic effects of paclitaxel (PA) and gemcitabine (GE) in vitro and in vivo on prostate cancer cell line PC-3. METHODS: Cell morphological observation, MTT assay, flow cytometry, and immunocytochemical method were used to observe the effects of 1+/-10(-6) mol/L, 1 x 10(-7) mol/L and 1 x 10(-8) mol/L PA and 1 x 10(-7) mol/L, 1 x 10(-8) mol/L and 1 x 10(-9) mol/L GE on prostate cancer cell line PC-3 in vitro in a single or combined administration for 48 h. Male BALB/C-nu mice bearing PC-3 prostate cancer were treated with docetaxol and retinoic acid singly or synergistically, followed by measurement of the body weight and immunohistochemical examination of serum prostate specific antigen (PSA) and PSA expression in the implanted tumors. RESULTS: GE at the concentration of 1 x 10(-8) mol/L significantly enhanced the effect of PA above 1 x 10(-7) mol/L in inducing growth inhibition (with an inhibition rate over 50.8%+/-4.2%, P<0.05) and apoptosis (apoptosis rate over 22.9%+/-2.3%, P<0.05) of PC-3 cells and in down-regulating the expression of cyclin D1 (expression rate no higher than 9.6%+/-1.6%, P<0.01) in PC-3 cells. GE lowered the rate of PA-induced cell cycle arrest at G(2)/M phase from 70.3%+/-9.7% to 38.2%+/-4.2%, and partially reversed the G(2)/M arrest (P<0.01). Synergistic treatment of the tumor-bearing mice caused little change in the body weight, but the serum PSA (51+/-14 ng/ml), implanted tumor mass (3.2+/-0.5 g) and PSA expression in the tumors (30%+/-3.7%) were all decreased significantly in comparison with the control mice (21.6+/-1.7 g). CONCLUSIONS: GE can enhance PA-induced tumor cell growth suppression and apoptosis in a synergistic manner both in vitro and in vivo, suggesting their great potential in clinical treatment of androgen-independent prostate cancer.

[Clinical study on application of bailing capsule after renal transplantation].

OBJECTIVE: To observe and assess the immunosuppressive effect of applying bailing capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia), after renal transplantation, its influence on other systems of organism, and to explore the possible therapeutic mechanism. METHODS: One hundred and twenty-one recipients of renal homo-allograft were randomly divided into two groups. The 64 cases in Group A was treated with cyclosporin A (Cs A) + prednisone (pred) + azathioprine (Aza), the 57 in Group B treated with Cs A + pred + BLC. They were followed-up for 1-2 year by checking up blood routine, urine routine, liver and renal function, blood electrolytes, glucose and lipids, and uric acid for 2 times every week in the first month after transplantation, followed by proper re-examination of these items according to various condition. RESULTS: There was no significant difference between the two groups in aspects of graft survival rate, occurrence of reject reaction, renal function recovery, blood electrolytes and blood glucose levels. However, as compared with Group A, in Group B, levels of urinary erythrocytes and leucocytes, blood alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol, uric acid as well as the incidence of infection were significantly lower, and blood high density lipoprotein, serum total protein, albumin, RBC and WBC count were significantly higher. CONCLUSION: BLC could effectively prevent the reject response after renal transplantation, protect renal and liver function, stimulate hemopoietic function, improve hypoproteinemia and hyperlipidemia, reduce the infection, etc., therefore, it is an ideal immunosuppressor after organ transplantation.

[Study on Toll-like receptors expression and cytokine production induced by bacillus Calmette-Guerin in human bladder cancer cell].

OBJECTIVE: To investigate the effect of bacillus Calmette-Guerin (BCG) on Toll-like receptors (TLRs) expression and cytokine production in human bladder cancer cell line T24. METHODS: Human transitional carcinoma cell line T24 was cultured in RPMI1640 medium with 10% FBS, BCG was added into the cell culture with various doses in bacteria-cell ratio. After T24 cells were stimulated by BCG for 1 hour, total RNA of cells was extracted. RT-PCR procedure was conducted with the primer of TLR-2 and TLR-4, and the products were analyzed with agarose gels electrophoresis. Then the expression of TLR-2 and TLR-4 in T24 cells was assessed by the analyzing system of gel imaging. After the stimulation culture for 12 hours, the supernatant of cells was collected. The levels of IL-4 and IL-12 in each sample were assayed by ELISA method. RESULTS: The expression level of either TLR-2 or TLR-4 was increased by the stimulation of BCG in a dose-dependent manner, the effects reached the maximal level at the dose of BCG as 10 bacilli per cell. The production of IL-12 in T24 cells was also increased gradually by the stimulation of BCG in a dose-dependent manner, and the dose of BCG obtained maximal effect was 10 bacilli per cell, which is coincident with the results observed in the expression of TLRs. There was no difference showed on the production of IL-4 between T24 cells stimulated with BCG and control. CONCLUSIONS: The stimulation of BCG not only up-regulated the expression of TLR-2 and TLR-4, but also increased the production of IL-12 in bladder cancer T24 cell line. The expression of TLRs and the production of cytokine in bladder cancer cells may be related to the BCG-induced immunol response to human bladder cancer.