Objective:To establish a method to determine benzoic in ibuprofen suspension. Methods:The determination of ben-zoic was performed on a Kromasil C18 (250 mm × 4. 6 mm, 5 μm) column with the mobile phase consisting of 0. 05 mol·L-1 monopo-tassium phosphate and acetonitrile (72 ∶28). The detection wavelength was 235 nm and the column temperature was 30℃. The injec-tion volume was 10μl. Results:The limit of detection was 3. 57 ng and benzoic had a good linear relationship within the range of 12. 5-200. 0 μg·ml-1(r=0. 999 9). The solution was stable in 24 hours. The average recovery was 98. 42% and RSD was 0. 98% (n=9) . Conclusion:The method is simple, repeatable and accurate, and can be used for the determination of benzoic in ibuprofen sus-pension.

Objective:To investigate the effect of intensive hypoglycemic therapy on the frequency of recurrent periodontitis in the patients with type 2 diabetes mellitus. Methods: Totally 40 patients with type 2 diabetes mellitus and recurrent periodontal disease were randomly divided into group A and group B. Group A was treated with the periodontal basic therapy combined with the convention-al hypoglycemic therapy. Group B was treated with the periodontal basic therapy combined with the intensive hypoglycemic therapy. Af-ter 6-month treatment,the change of the probing depth of the periodontal pocket, sulcus bleeding index, incidence frequency, recovery course, body mass index, fasting blood glucose, glycosylated hemoglobin and serum C-reactive protein levels were measured before and after treatment. Results:After the treatment, the indices were improved in group A except body mass index and fasting blood glucose (P<0. 05), and in group B, the indices were improved except body mass index (P<0. 05), and the probing depth of the periodontal pocket, sulcus bleeding index, incidence frequency, body mass index, serum C-reactive protein levels, glycosylated hemoglobin and fasting blood glucose of the patients in group B were all better than those in group A (P<0. 05). Conclusion:Periodontal basic thera-py combined with intensive hypoglycemic therapy can effectively improve the periodontal health of diabetic patients, shorten the recov-ery treatment of periodontal disease, reduce the incidence frequency, and reduce blood glucose as well.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

ObjectiveTo observe the effect of ginsenosides Rb1 on cerebral blood flow of rat models with cerebral ischemia/reperfusion (I/R) injury, which could provide a new theory of cerebral protective mechanism about ginsenosides Rb1.Methods Twenty-four rats were randomly divided into sham-operation group, model group, normal saline control group and ginsenosides Rb1 group, 6 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by thread embolism method. At the end of I/R, in the rat of ginsenosides Rb1 group, ginsenosides Rb1 40 mg/kg was immediately intraperitoneally injected, while in the rat of normal saline control group, an equal volume of normal saline was injected intraperitoneally. After I/R for 24 hours, the cerebral local amount of blood flow was measured, the rats' behavior score was observed, and the volume of cerebral infarction was monitored by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining.Results The percentage of volume of cerebral infarction [(64.23±8.12)% vs. 0%] and behavior score [3.0 (2.0-4.0) vs. 0 (0-0),P< 0.05] in model group were significantly higher than those in sham-operation group, while the cerebral local amount of blood flow in model group was obviously lower than that in sham-operation group (mL/min: 125.75±57.65 vs. 225.01±78.25,P< 0.05); Compared with the model group and normal saline control group, the percentage of volume of cerebral infarction [(23.62±8.74)% vs. (64.23±8.12)%, 56.72±8.92] and behavior score [0.5 (0.0-2.0) vs. 3.0 (2.0-4.0), 3.5 (1.0-4.0)] in the ginsenosides Rb1 group were significantly lower, the cerebral local amount of blood flow was markedly increased in the ginsenosides Rb1 group (177.25±75.36 vs. 125.75±57.65, 132.65±58.65,P< 0.05).Conclusion Ginsenosides Rb1 can increase the cerebral blood flow in rats with cerebral I/R injury, which maybe one of the mechanisms of cerebral protection of Ginsenosides Rb1.

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

Objective To investigate the effect of prostaglandin E: (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than these of the control group (P<0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P>0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.

The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows： 〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%） （P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%） （P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.

Objective To study the safety and efficacy of damage control using percutaneous transhepatic biliary drainage (PTBD) in acute cholangitis of severe type (ACST) secondary to intrahepatic choledocholithiasis.Methods The clinical data of 8 patients who received PTBD after hospital admission followed by conventional surgery for ACST when their general condition improved were retrospectively studied.Results All patients received PTBD successfully and the amount of bile drained was 100-400 ml in the first day.The general condition of these 8 patients became better after 24 h and the total bilirubin decreased for about 25-100 mmol/L after 48 h.Three patients with a platelet count of less than 20 × 109/L showed an improved count to more than 50 × 109/L 72 h after PTBD.All patients were operated at different times after the PTBD:2 received T-tube drainage,3 T-tube drainage combined with left hepatectomy,and 3 choledochojejunostomy.Seven patients recovered uneventfully,but 1 developed hepatic failure with the total billurubin rose to more than 200 μmol/L.He was discharged home with the PTBD tube.During the waiting time of 7 days to 3 months before surgery,the tubes were kept patent and no mortality or morbidity such as bleeding,bile leakage,and peritonitis occurred.Conclusions PTBD was a safe and efficacious procedure for patients who were in a serious condition with ACST secondary to intrahepatic choledocholithiasis.It was more likely to be successful as it is minimally invasive and therefore well-tolerented.It reduced the biliary pressure,relieved the ongoing sepsis,and was a good preparatory procedure before any conventional surgery.

Objective To investigate the clinical value of color Doppler flow imaging (CDFI) for evaluating transplant renal artery stenosis (TRAS).Methods Clinical and ultrasonographic data of 216 kidney transplant recipients were collected by follow-up monitoring from September 2015 to July 2016.CDFI indexes included the peak systolic velocity (PSV) in the renal artery and resistant index (RI).Renal artery PSV and RI were measured.All suspected TRAS patients accepted transplant renal artery angiography (DSA).Results Fourteen patients with suspected TRAS accepted DSA,of which 12 patients were confirmed.The diagnostic accuracy of CDFI was 85.7％.When the POST-PSV ratio＞ 1 0,the sensitivity and specificity of diagnosis of TRAS were 91 ％ and 95 ％,respectively.CDFI indexes remarkably changed after the TRAS patients had undergone renal artery dilatation or stent implantation.PSV of the main renal artery and the POST-PSV ratio decreased significantly,and the PSV of interlobar arteries increased.Conclusions CDFI is a reliable first choice for screening transplant renal artery stenosis.The POST-PSV ratio has relatively higher sensitivity and specificity in the diagnosis of TRAS.

The feasibility and the clinical value of the enzyme-multiplied immunoassay technique (EMIT) monitoring of blood concentrations of cyclosporine A (CsA) in patients treated with CsA were investigated after kidney transplantation.The validation method was performed to the EMIT determination of CsA blood concentration,the CsA whole blood trough concentrations (Co) of patients in different time periods after renal transplantation were monitored,and combined with the clinical complications,the statistical results were analyzed and compared.EMIT was precise,accurate and stable,also with a high quality control.The mean postoperative blood concentration of CsA was as follows:＜1 month,(281.4± 57.9)ng/mL; 2 - 3 months,(264.5 ± 41.2)ng/mL; 4 - 5 months,(236.4 ± 38.9)ng/mL; 6 - 12 months,(206.5 ± 32.6)ng/mL; ＞12 months,(185.6 ± 28.1)ng/mL.The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1％,significantly lower than that of the none-recommended dose group (37.2％) (P＜0.05); the transplantation rejection rate was 4.4％,significantly lower than that of the nonerecommended dose group (22.5％) (P＜0.05).Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible,and is able to reduce the CsA toxicity and rejection significantly,leading to achieving the desired therapeutic effect.