OBJECTIVE:To establish the method for simultaneous determination of 9 common inorganic anions in Huanglian shangqing tablet. METHODS:The accelerated solvent extraction-ion chromatography was adopted. Inorganic anions were deter-mined by Ion Pac AS11-HC anion exchange column,protected by Ion Pac AG11-HC column and eluted by hydroxide potassium so-lution(gradient elution)at the flow rate of 1.2 mL/min. The column temperature was set at 30 ℃,elution time was 18 min,and sample volume was 25 mL. RESULTS:The linear ranges of fluorinion,formate ion,nitrite ion,bromide ion,nitrate ion,sulfate ion,oxalate ion and phosphate ion were 0.1-5 mg/L(r=0.9990-0.9999). The limits of quantitation were 0.020,0.078,0.030, 0.058,0.052,0.068,0.084,0.064,0.074 mg/L,and the limits of detection were 0.005,0.024,0.008,0.017,0.015,0.022, 0.026,0.020,0.021 mg/L,respectively. RSDs of precision,stability and repeatability were all lower than 4.0%;recoveries were 80.00%-125.08%(RSD ranged 0.97%-2.47%). CONCLUSIONS:The method is simple,precise,stable and repeatable,and can be used for 9 common inorganic anions in Huanglian shangqing tablet.

BACKGROUND: To prepare a novel biological material that can be implanted into the lesion of bone tuberculosis, keep sustained release of anti-tuberculosis drugs around bone tuberculosis tissues for a long time, and enhance the effect on bone repair has become a hot spot for clinical studies on bone tuberculosis.OBJECTIVE: To prepare the adipose-derived mesenchymal stem cells (ADMSCs)/sustained-release rifampin-loaded microsphere complex, and to preliminarily study its effects in a rabbit model of spinal tuberculosis.METHODS: Rifampin-chitosan-calcium alginate sustained-release microspheres were synthesized, and co-cultured with ADMSCs to prepare an anti-tuberculosis composite for bone tissue engineering. Forty New Zealand rabbits were used to make lumbar L6 tuberculosis models. Thereafter, rat models were randomly divided into four groups and given rifampin by gavage in rifampin group, ADMSCs by paravertebral injection+rifampin by gavage in stem cell group, rifampin by gavage+implantation of the anti-tuberculosis composite material in experimental group, and no treatment in control group.The duration time of rifampin administration was 8 weeks. The anti-tuberculosis effect of the composite material was evaluated by X-ray and CT scanning observation.RESULTS AND CONCLUSION: In the control group, obvious damage to lumbar vertebrae L5 and L6 was apparent; inflammatory granulation tissues formed; and the intervertebral space was narrowed. In addition, two rabbits in the control group showed obvious kyphotic deformity and five showed pasoas major swelling with low-density dark region in the psoas muscle. In the rifampin group, there were five rabbits with moderate damage of the lumbar vertebrae L5 and L6,and two rabbits with pasoas major swelling. In the stem cell group, there were two rabbits with moderate damage of the lumbar vertebrae L5 and L6, three rabbits with mild damage of the upper part of the lumbar vertebra L6, and three rabbits with pasoas major swelling. In the experimental group, only four rabbits suffered from mild damage of the upper part of the lumbar vertebra L6 but with no changes in the intervertebral space between the L5 and L6, and without pasoas major swelling. These results indicate that the composite material of ADMSCs combined with sustained-release rifampin-loaded microspheres can inhibit mycobacterial growth effectively, and reduce vertebral bone destruction,thereby giving some therapeutic actions for the animal models with spinal tuberculosis.

AIM: To study the expression of the VEGF in single and multiple superficial transitional cell bladder carcinoma and their clinical significance. METHODS: Immunohistochemical method was used to study the VEGF in 60 cases of superficial transitional cell bladder carcinoma and in 10 cases of normal bladder tissue as control. RESULTS: High expression of VEGF in bladder carcinoma cell was observed. The expression level of VEGF in multiple superficial transitional cell bladder carcinoma was higher than that in single superficial transitional cell bladder carcinoma. The recurrent rate in the patient with VEGF high expression was more than that in the patient with VEGF low expression. CONCLUSION: The expression level of VEGF was correlated to the biological behavior of superficial transitional cell bladder carcinoma.

Objective To discuss the blockage reasons and countermeasures of temporary catheter in central venous including femoral vein and internal jugular vein for hemodialysis patients.Methods The reasons of blockage of temporary catheter in central venous among 60 hemodialysis patients from January 2014 to September 2015 in Hemodialysis Room of the First People's Hospital of Longquanyi district of Chengdu were reviewed.There were 32 cases in femoral vein and 28 cases in the right internal jugular vein.The incidence of blockage of the two temporary catheter in the first two weeks,the incidence of blockage caused by mechanical factors and blood clots or fibrous sheath and the incidence of blockage which needed to change the catheter in the two groups patients within the first two months were analyzed.Results The first rate of defective function in the first two weeks of femoral vein group was 53.13％(17/32),and 7.14％(2/28) of the right internal jugular vein group,there were significant difference between the two groups (x2=13.061,P＜0.001).The incidence of catheter blockage due to mechanical factors of femoral vein group was 96.88％ (31/32),and 53.57％ (15/28) of the right internal jugular vein,there were significant difference between the two groups (x2 =15.654,P＜0.001).The incidence of blockage caused by blood clots or fibrous sheath and the incidence of blockage which needed to change the catheter in the first two months were no significant differences between the two groups(P=0.061,0.096).Conclusion The cases of temporary catheter in femoral vein have blockage earlier,and they have a higher risk of catheter jam than the cases of temporary catheter in right internal jugular vein.

BACKGROUND:Smooth muscle cells and transitional epithelial cells were traditionally used to construct tissue-engineered bladder and to perform double-sided implantation of scaffold.However,double-sided implantation is difficult to perform,because smooth muscle cells are difficult to isolate or culture in vitro and passage is limited.OBJECTIVE:To verify the feasibility of tissue-engineered bladder reconstruction with bone marrow mesenchymal stem cells(BMSCs)and bladder acellular matrix(BAM).DESIGN:A basic empirical study.SETTING:Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University.MATERIALS:Experiments were performed at the Linbaixin Medical Research Center,Second Affiliated Hospital,Sun Yat-sen University from March 2006 to Mav 2007.The laboratory was the Opening Laboratory of Hospital Affiliated to Health Department of China.One-month old SD rats of either sex,weighting 80-100 g were provided by Animal Experimental Center of Sun Yat-sen University.Fresh porcine bladders were offered by Animal Experimental Center of Southern Medical University.METHODS:Whole bone marrow culture and successive adherence method was used to culture rat BMSCs in vitro.Flow cytometry was employed to detect surface antigen.Eradicator washing method was applied to prepare porcine BAM and measure its purity and characteristies.Third passage of BMSCs were inoculated in BAM and cultured in a medium containing vascular endothelial growth factor(VEGF165)(25 ng/L)in vive and in vitro to test compatibility.Cells cultured alone were considered to be controls for the in vivo trial,and materials non-implanted with cells were considered to be controls for in vitro trial.Suitable microenvironment was simulated to induce the differentiation of BMSCs.Four weeks and eight weeks later,compound materials were respectively removed to perform tissue section test.Simultaneously,immunohistochemistry keratin staining was conducted to examine regeneration of epithelial cells.MAIN OUTCOME MEASURE:Biocompatibility of BMSCs and BAM.RESULTS:①BMSCs were cultured by whole bone marrow method.Flow cytometry demonstrated that third passage of cells were positive for CD29(99.43%).②BAM had good biological characteristics.Homogen matrix and byssoid collagen appeared under a microscope.Compatibility trials showed good compatibility of BMSCs and BAM and well-growth cells.③Four weeks later,histological section test confirmed inflammatory cell infiltration,closely-arranged collagen and elastic fiber.Immunohistochemistry keratin staining showed lamellar and discontinuous simple epithelium.Eight weeks later,no inflammatory cell infiltration was found,and closely-arranged collagen and elastic fiber were detected.Immunohistochemistry keratin staining showed lamellar and continuous multiple epitheliums.CONCLUSIoN:With good compatibility,BMSCs and BAM appear to be an ideal material for bladder tissue engineering.

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.

AIM: To investigate serum vascular endothelial growth factor (VEGF) level in patients with bladder transitional cell carcinoma(BTCC) and its clinical significance. METHODS: ELISA method was used to examine the serum VEGF level in 42 cases of bladder transitional cell carcinoma and in 10 cases of normal people as control. The change of VEGF in blood of the pre-operation and post-operation patients with BTCC was also compared. RESULTS: The VEGF level in blood of the patients was higher than that of the normal people, in spite of pre-operation, post-chemotherapy, and post-operation, but VEGF level decreased obviously after chemotherapy or operation. In addition, the plasma VEGF level was related to the grade and invasion of tumor. CONCLUSION: Detecting serum VEGF level can help us to assess the change of tumor and therapeutic effect.

Acute kidney injury(AKI)is a very common critical disease worldwide, especially in pediatric intensive care unit(PICU). There are many pathogenic factors for AKI, among which sepsis is essential to the development of AKI.Sepsis-associated acute kidney injury(SA-AKI) often have sudden onset and poor prognosis.Because there is still a lack of powerful diagnostic tools, resulting in clinical intervention in SA-AKI often lags behind.Therefore, awareness of SA-AKI risk and early identification of injury severity, as well as timely initiation of supportive treatment, remain the main contents of current research.There are hundreds of new biological markers and diagnostic tools for early identification and prognosis of AKI.This review introduced the progress on diagnosis of SA-AKI.The aim is to improve clinicians′ understanding of SA-AKI.

Purpose：To study the expressions of transforming growth factor beta 1(TGFβ1) and proliferating cell nuclear antigen(PCNA) and their effects on the growth of human renal cell carcinoma.Methods:46 specimens of human renal cell carcinoma (RCC) and 11 normal kidneys as control were assessed by immunohistochemical method (SP method).Results:TGFβ1 content in RCCs was more than that in normal kidneys (P<0.05). The PCNA index in RCCs was higher than that in normal kidneys (P<0.01). The PCNA index was significantly higher in poor differentiated and invasive RCCs (P<0.05). There was a negative relationship between the content of TGFβ1 and PCNA（P<0.01）.The expected life was shorter in RCC cases with low level TGFβ1 or high level PCNA(P<0.05).Conclusions:TGFβ1 can act as a negative regulating factor and inhibit the growth of RCC, especially inhibit the proliferation of RCCs. TGFβ1 and PCNA can act as the predict factors of the progression of RCCs.

Objective To study the feasibility and safety of performing nephrectomy together with the removal of complicated inferior vena cava tumor thrombus under profound hypothermia and arrested circulation. Methods After made the median thoraco-abdominal incision, the exploration of the abdominal organs was done. The right kidney, inferior vena cava and renal pedicle were well exposed then. After the whole body heparinization, cannulas were put into ascending aorta, superior vena cava, aortic root and right superior pulmonary vein. The body temperature was reduced to 20℃ with cardiopulmonary bypass unit and the extracorporeal circulation was stopped then. Cut open the inferior vena cava at vena renalis dextra ingress and the F16 urinary catheter was inserted into atrum dextra through inferior vena cava and inflated. The tumor thrombus was pulled out and the right kidney was removed. The inferior vena cava incision was sutured to close and the extracorporeal circulation was resumed and patient was re-warmed.Results The operation time was 330 min and the extracorporeal circulation time was 90 min, while the profound hypothermia with circulatory arrest time was 20 min. The estimated blood loss during operation was 400 ml and 6 unit red cells and 600 ml blood plasm were transfused. The patient was awaked 2.5 h after the operation, food intake resumed 4 days after operation and the patient was discharged on day 10 post-operatively. After 6 months'follow-up, there were no local recurrence and metastasis occurred. Conclusion The technique of profound hypothermia and circulation arrest could improve the safety and efficacy in the treatment of renal cell carcinoma with suprahepatic (level Ⅲ) caval tumor thrombus.