BACKGROUND: High oxalic acid urine is a risk factor for stone formation. Constructing cel lines with high oxalate metabolic ability using genetic engineering and stem cel technology wil become the effective method to prevent and treat calcium oxalate stone. OBJECTIVE: To construct the adult bone marrow mesenchymal stem cel lines that can decompose the oxalic acid, through co-transfecting the oxalic acid degradation genes Frc and Oxc of oxalobacter formigenes into the normal adult bone marrow mesenchymal stem cells. METHODS: Frc and Oxc were amplified by PCR, and the eukaryotic expression vectors of pLEGFP-N1-myc-Frc and pBaBE-puro-flag-Oxc were constructed, then co-transfected into the normal adult bone marrow mesenchymal stem cells. The non-transfected bone marrow mesenchymal stem cells and the cells transfected with empty vectors were as control. Western blot was performed to detect the expression of the objective genes;the concentration of oxalate in the culture medium after transgenic was determined by ion chromatography. RESULTS AND CONCLUSION: Restriction enzyme digestion and sequencing results showed that the Frc and Oxc genes were successful y amplified, and the vectors of pLEGFP-N1-myc-Frc and pBaBE-puro-flag-Oxc were constructed. After tranfected into the bone marrow mesenchymal stem cells, the Western blot results showed that transfected bone marrow mesenchymal stem cells could stably express the target protein myc-formyl coenzyme A transferase enzyme and the flag-oxalyl coenzyme A decarboxylase; ion chromatography test results showed with the prolonging of the culture time, the concentration of oxalic acid in the human bone marrow mesenchymal stem cel culture medium transfected with target gene was decreased gradual y. While there was no target protein expression in the non-transfected human bone marrow mesenchymal stem cells as wel as the cells transfected with empty vectors. The cells had the ability of oxalate-degradation. The experiment successful y constructs the adult bone marrow mesenchymal stem cel lines that can decompose the oxalic acid, and the cel lines have the ability of oxalate-degradation and can stably express the oxalate decomposition proteins Frc and Oxc.

A HPLC-MS/MS multiple-reaction monitoring (MRM) quantitative analysis was made to establish a determination method for drug concentrations of costunolide (Co) and dehydrocostuslactone (De) in blood samples in the positive ion mode, with diazepam as the internal standard substance, in order to study the pharmacokinetic process of sesquiterpene lactones costunolide and dehydrocostuslactone after the oral administration of Weichang'an pills, and provide an theoretical basis for further studies on the substance basis for the anti-diarrhea effect of Weichang'an pills. In the blood samples, Co and De showed a good linearity within concentration ranges 0.700 0-769.7, 2.510-956.0 μg x L(-1), respectively. The results of precision, stability and recovery experiences proved the stability and reliability of the plasma concentration determination method. After the oral administration, the concentrations of Co and De in plasma increased with the increase in dose, with T(max) between 10.65-12.98 h, indicating a long time to reach peak plasma concentrations; C(max) of costunolide and dehydrocostuslactone ranged between 3.750-5.450,15.34-44.52 μg x L(-1), respectively. The in vivo adsorption of Co and De conformed to the one-compartment model, with a longer time to attain the peak plasma concentrations. These results provided an experimental basis for revealing the active substance basis and clinical medication of Weichang'an pills.
Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Lactones ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Wistar ; Sesquiterpenes ; administration & dosage ; blood ; pharmacokinetics ; Tablets ; administration & dosage ; chemistry ; pharmacokinetics

Objective To review the safety and efficacy of upper-pole access percutaneous nephrolithotomy (PCNL). Methods From May 2008 to May 2010,237 cases ( 135 males and 102 females with mean age of 42 yrs) of renal or proximal ureteral calculi treated with upper-pole access PCNL were reviewed.The indications included calculus larger than 1.5 cm or impacted proximal ureteral calculi in 94 cases,calculus≥2 cm or impacted renal pelvic calculi in 26 cases,staghorn or multiple calculi in 68 cases,complex lower calyx calculi in 13 cases,upper calyx calculi not amenable to ESWL or URS in 12 cases,calculi within upper calyx diverticulum in 3 cases,combined UPJ obstruction or upper ureteral stenosis in 8 cases,morbidly obese patients in 3 cases,calculi within horseshoe kidneys in 6 cases,calculi within transplanted kidneys or ureters in 4 cases.Of the 237 cases,175 tracts (73.8％) were above the 12th rib,46were above the 11th rib,12 were below the 12th rib,4 were in the lower abdomen for renal transplant patients. Results The overall stone clearance rate with upper-pole access PCNL monotherapy was 74.3％.Additional punctures were required in 55 cases and combined ESWL in 6 cases.Total stone clearance rate at 3 months after operation was 88.2％.16 patients (6.8％) had a pleural injury.Thoracentesis was required in 8 patients,closed thoracic drainage in 5 patients,conservative treatment in 3 patients.After nephrostomy tube removed,pleural irritation symptom appeared in 12 cases (5.1％) who required symptomatic treatment.No patient had injury to the lung or other viscera.Significant bleeding requiring blood transfusion was olserved in 5 patients,while selective renal arterial embolization was required in 2 cases. Conclusions Upper-pole access offers optimal visibility,convenience for the movement of rigid nephroscope and high stone-free rate.It should be attempted in selected cages of upper ureteral calculi and complex renal calculi.

Objective To evaluate the efficacy and safety of bladder hydrodistention for the treatment of ketamine-associated bladder dysfunction.Methods Six patients were required to withdraw the ketamine and treated with bladder hydrodistention therapy and sodium hyaluronate irrigation,and medicine to pretect liver and kidney was also used.Results The biopsies of 6 cases demonstrated the cystitis through biopsy.Lower urinary tract symptoms such as urgency,thamuria and odynuria were significantly relieved after bladder installation within 30 days.The O'Leary-Sant ICSI scores and the ICPI scores reduced to 3.5 ± 1.6,2.8 ± 1.5 respectively.The functional bladder capacities increased to an anverage of (180 ± 28)ml,.2-3 times of nocturia,Qmax (14.4 ± 4.3) ml/s.All cases were followed up for 4 to 18 months.Symptoms disappeared or were significantly relieved in all patients.Conclusion Contracture of bladder might be the main presentation of ketamine-associated bladder dysfunction.Intravesical hydrodistention therapy and sodium hyaluronate irrigation could be the safe and effective therapy in the treatment of katamine-associated dysfunction.

Objective To discuss the diagnosis and treatment of renal pelvic carcinoma associated with renal stone.Methods A total of 13 patients,aged from 49 to 73 years old and averaged 59years old.The history of renal stone was 16 years.13 patients accepted B ultrasound check and 1 was found soft tissue occupying.8 patients accepted IVU check and none was found soft tissue occupying.7 patients accepted CT scan and 4 were found soft tissue occupying.The fluorescence in situ hybridization (FISH) examination was done for 2 patients and both were positive.6 patients were found lesions at renal pelvis mucous membrane during the operation of percutaneous nephrolithotripsy,4 accepted radical operations of renal pelvic carcinoma and 2 patients accepted radical nephrectomy according to the biopsy pathology.4 were found soft tissue occupying before operation and accepted radical operation of renal pelvic carcinoma ultimately.1 patient suffered gross hematuria and renal insufficiency accepted the renal pelvic carcinoma vaporization under the ureteroscope.Results The pathology showed that 7 cases were transitional cell carcinoma,4 were transitional cell carcinoma combined squamous cell carcinoma (SCC) metaplasia and 2 were squamous cell carcinoma.6 of 8 patients' stone chemical composition were infection stone and 2 were calcic blended stone.3 patients were followed up 1 to 2 years and survival with no tumor recurrence.The tumor recurred 10 months of the patient accepted the operation of renal pelvic carcinoma vaporization and accepted vaporization again.1 patient bsuffered SCC and local lymph node metastasis.He died 13 months post-operation.Conclusions For the patient who had long history of stone,combining infection with symptoms of severe hematuria and postoperation hematuria,the possibility of renal pelvic carcinoma should be considered.CT scan and urine FISH may help for diagnosis.The biopsy should routinely perform for the doubtful mucosa lesion during the cavity stone operation.Early and timely diagnosis and operation is the key for the patients with pelvic carcinoma associated with renal stone.

Some unhealthy life habits, such as long-term smoking, heavy drinking, sexual overstrain and frequent stay-up could induce the Yin deficiency symptoms of zygomatic red and dysphoria. Stems of Dendrobii officinalis flos (DOF) showed the efficacy of nourishing Yin. In this study, the hyperthyroidism Yin deficiency model was set up to study the yin nourishing effect and action mechanism of DOF, in order to provide the pharmacological basis for developing DOF resources and decreasing resource wastes. ICR mice were divided into five groups: the normal control group, the model control group, the positive control group and DOF extract groups (6.4 g · kg(-1)). Except for the normal group, the other groups were administrated with thyroxine for 30 d to set up the hyperthyroidism yin deficiency model. At the same time, the other groups were administrated with the corresponding drugs for 30 d. After administration for 4 weeks, the signs (facial temperature, pain domain, heart rate and autonomic activity) in mice were measured, and the facial and ear micro-circulation blood flow were detected by laser Doppler technology. After the last administration, all mice were fasted for 12 hours, blood were collected from their orbits, and serum were separated to detect AST, ALT, TG and TP by the automatic biochemistry analyzer and test T3, T4 and TSH levels by ELISA. (1) Compared with the normal control group, the model control group showed significant increases in facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01), serum AST, ALT (P < 0.01), T3 level (P < 0.05), TSH level (P < 0.05) and notable deceases in pain domain (P < 0.01), TG level (P < 0.01). (2) Compared with the model control group, extracts from DOF (6 g · kg(-1)) could notably reduce facial and ear micro-circulation blood flow, facial temperature and heart rate (P < 0.05, P < 0.01) and AST (P < 0.05) and enhance pain domain (P < 0.01) and TG (P < 0.01). Extracts from DOF (4 g · kg(-1)) could remarkably reduce AST and ALT levels (P < 0.01, 0.05). Extracts from DOF (6 g · kg(-1) 4 g · kg(-1)) could significantly reduce T3 and increase serum TSH level (P < 0.05). DOF could improve Yin deficiency symptoms of zygomatic red and dysphoria in mice as well as liver function injury caused by overactive thyroid axis. According to its action mechanism, DOF may show yin nourishing and hepatic protective effects by impacting thyroxin substance metabolism, improving micro-circulation and reducing heart rate.
Animals ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flowers ; chemistry ; Humans ; Hyperthyroidism ; drug therapy ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Thyroxine ; metabolism ; Yin Deficiency ; drug therapy ; metabolism

OBJECTIVETo investigate the influence of myocardial inhibition on injury to liver, kidney and intestine and blood perfusion at early stage in rat with severe burn.

METHODSThirty-two healthy male Wistar rats were enrolled in the study and randomly divided into sham scald, propranolol, scald control and scald + propranolol groups, with 8 rats in each group. After intraperitoneal injection of 10 g/L pentobarbital sodium for anesthesia, rats of the former two groups were sham scalded in a water bath of 37 degrees C for 18 s, while the latter two groups were inflicted with 30% TBSA full-thickness scald in a 97 degrees C water bath for 18 s. Rats were resuscitated with Ringer's lactate solution (4 mL x kg(-1) x 1% TBSA(-1), i. p.) following the Parkland formula 30 mins after the injury. At the same time, rats in propranolol and scald + propranolol groups received propranolol 0.75 mg/kg intravenously. After 6 hours, parameters of myocardiac mechanics (SBP, DBP, MAP, LVSP, LVEDP, +/- dp/dt max) were recorded by the multiple channel physiological signal collecting and processing system; blood flow of liver, kidney and intestine were detected with the laser doppler flowmetry; the serum contents of cTnI, TBA, beta2-MG and DAO were determined for reflecting injuries to the heart, liver, kidney and intestine, respectively.

RESULTSMyocardiac mechanics parameters, with the exception of LVEDP, were decreased in propranolol group as compared with those in sham group (P <.05). All myocardiac mechanics parameters in burn control group were lower than those in sham group and higher than those in burn + propranolol group (P < 0.05). Blood flow of organs showed similar changes in above-mentioned 3 groups (P < 0.05). Organ damages as shown in burn control group [cTnI (4.86 +/- 0.29) microg/L, TBA (83.6 +/- 18.2) micromol/L, beta2-MG (2.75 +/- 0.19) mg/L, DAO (1.45 +/- 0.09) x 10(3) U/L] were more serious than those in sham control group [cTnI (1.73 +/- 0.09) microg/L, TBA (24.5 +/- 2.4) micromol/L, beta2-MG (1.15 +/- 0.18) mg/L, DAO (0.87 +/- 0.13) x 10(3) U/L], and less serious than those in scald + propranolol group [cTnI 5.95 +/- 0.42 microg/L, TBA 125.8 +/- 21.3 micromol/L, beta2-MG 3.25 +/- 0.17 mg/L, DAO (1.83 +/- 0.13) x 10(3) U/L] (P < 0.05).

CONCLUSIONSPropranolol can aggravate injury to the liver, kidney and intestine at early stage in rat with severe burn, suggesting that "shock heart" may be one of initial factors in lowering blood flow to the organs, thus inducing injury to them.

Animals ; Blood Pressure ; Burns ; metabolism ; physiopathology ; Intestines ; blood supply ; Kidney ; blood supply ; Liver ; blood supply ; Male ; Myocardium ; metabolism ; Propranolol ; adverse effects ; Rats ; Rats, Wistar ; Shock

Ursolic acid (UA) is a sort of pentacyclic triterpenoid carboxylic acid purified from natural plant. UA has a series of biological effects such as sedative, anti-inflammatory, anti-bacterial, anti-diabetic, antiulcer, etc. It is discovered that UA has a broad-spectrum anti-tumor effect in recent years, which has attracted more and more scholars' attention. This review explained anti-tumor actions of UA, including (1) the protection of cells' DNA from different damages; (2) the anti-tumor cell proliferation by the inhibition of epidermal growth factor receptor/mitogen-activated protein kinase signal or of FoxM1 transcription factors, respectively; (3) antiangiogenesis, (4) the immunological surveillance to tumors; (5) the inhibition of tumor cell migration and invasion; (6) the effect of UA on caspase, cytochromes C, nuclear factor kappa B, cyclooxygenase, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) or mammalian target of rapamycin signal to induce tumor cell apoptosis respectively, and etc. Moreover, UA has selective toxicity to tumor cells, basically no effect on normal cells. With further studies, UA would be one of the potential anti-tumor agents.
Angiogenesis Inhibitors ; pharmacology ; therapeutic use ; Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Humans ; Immunologic Surveillance ; drug effects ; Neoplasms ; blood supply ; drug therapy ; immunology ; pathology ; Triterpenes ; chemistry ; pharmacology ; therapeutic use

OBJECTIVE: To compare the flow-mediated dilatation (FMD) among the newly diagnosed impaired glucose tolerance (IGT), type 2 diabetes mellitus (T2DM), and the normal controls (NC) and to analyze relevant factors under different glucose levels. METHODS: The study included IGT (n=34), DM1 (n=52), DM2 (n=33) and NC (n=25). Levels of fasting plasma glucose (FPG), 2-hour postprandial plasma glucose (PPG), fasting insulin (FINS), 2-hour postprandial insulin (PINS), triglyceride (TG), total cholesterol (TC), and hemoglobin A1C (HbA1C) were determined in all participants. High resolution ultrasound examining FMD was performed to measure vascular endothelial function subsequently. RESULTS: There was statistically significant difference between IGT, DM, HG and NC group in FMD (P=0.008). Partial correlation analysis found that a significant negative correlation existed between FMD and homeostasis model assessment-index (HOMA-IRI), difference of plasma glucose (DPG), FPG and PPG (P<0.01), and a negative correlation between FMD and HbA1C (P<0.05). Setting FMD as dependent variable to conduct multiple linear stepwise regression, in IGT group it was the waist/hip ratio (WHR) and HOMA-IRI that entered the regression equation; in DM1 group it was HOMA-IRI, PPG and DPG that entered the regression equation; in DM2 group it was FPG and HOMA-beta that entered the regression equation. CONCLUSION There exists a flow-mediated vasodilatation dysfunction in patients of newly diagnosed IGT and T2DM. Effect of relevant factors on FMD differs with different glucose levels.
Adult ; Blood Glucose ; metabolism ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; blood ; physiopathology ; Endothelium, Vascular ; physiopathology ; Female ; Glucose Intolerance ; blood ; physiopathology ; Humans ; Male ; Middle Aged

Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; SARS Virus ; immunology ; isolation & purification ; Seroepidemiologic Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; virology