AIM: To investigate the effect of yigu capsule drug-containing serum on proliferation, ALP activity and expression of interleukin-11 mRNA in Sprague-Dawley rat osteobalsts in the co-culture system. METHODS: (1) Osteoblasts and osteoclasts were isolated from 1 and 5-day-old Sprague-Dawley rats, respectively. The osteoblasts-osteoclasts co-culture system was built to prevent the two kinds of cells from contact and allow the media to exchange. The experiment included two groups, drug-containing sera group and control group. (2) The osteoblasts proliferation, ALP activity and expression of interleukin-11 mRNA were detected by the MTT, 4-aminoantipyrine spectrometric methods and FQ-PCR,respectively. RESULTS: In drug-containing sera group, the osteoblasts proliferation, ALP activity, expression of IL-11 mRNA were higher than those of control group (P

AIM:To observe the effect of TNF-? and Yigu capsule drug-containing sera on osteoblasts apoptosis in osteoblasts-osteoclasts coculture system.METHODS:(1)Twenty female Sprague-Dawley rats,10 months old,were randomly assigned into 2 groups,NS and Yigu capsule groups,to prepare blank sera and drug-containing sera.(2)The DNA gel electrophoresis method was used to detect apoptosis of osteoblasts treated with different concentration of TNF-? in order to determine the best dosage in the co-culture system.(3)The cells were divided into four groups,TNF-? group,normal group,TNF-? + blank serum group and TNF-? + drug-containing serum group.DNA gel electrophoresis,acridine orange staining and flow cytometry were used to observe osteoblast apoptosis in these groups.RESULTS:(1)After induced by TNF-? for 72 h at a concentration of 60 ?g/L,relatively typical DNA ladder appeared in TNF-? group.(2)Only two DNA brands appeared and most of cells were well-proportioned stained in TNF-? + drug-containing serum group,the rate of osteoblasts apoptosis in TNF-? + drug-containing serum group(9.60%?0.26%)was obviously lower than that in the TNF-? group(26.90%?0.06%)and TNF-? + blank serum group(18.10%?0.06%).CONCLUSION:TNF-? induces osteoblast apoptosis in the co-culture system,and Yigu capsule drug-containing serum prevents osteoblast apoptosis induced by TNF-?.

Objective To observe the effect of long-term intensive glucose control therapy on diabetic retinopathy in outpatients with type 2 diabetes mellitus.Methods Forty-nine patients with type 2 diabetes mellitus were randomly assigned to participate in the trial from 2002 to 2007,receiving either intensive (24 cases) or standard glucose control (25 cases).The patients were examined by the same ophthalmologists to identify any new diabetic retinopathy (DR).After 5 years of intensive glucose control,all of the patients were asked to attend our clinic every 6 months,but no attempts were made to maintain their previously assigned therapies.Hemoglobin A1c (HbA1 c) was measured regularly.In November of 2009,a retinal examination was carried out by the same ophthalmologist who worked in the trial.The visual acuity,lens,vitreous body and fundus were examined after pupil dilation to identify diabetic retinopathy (DR).Fundus fluorescein angiography and retinal laser photocoagulation were carried out when necessary.Results In the second year after enrollment in the trail,the median HbA1c level of the intensive-therapy group was significantly lower than that of the standard-therapy group [(6.3 ± 0.6) ％ vs.(7.2 ± 1.2) ％,t =2.09,P ＜ 0.05],and was maintained in a controlled level throughout the following 4 years.During the post-trial monitoring,no new case of of macula edema or diabetic associated blindness occurred in either intensive or standard-therapy group,the whole occurrence of micro-aneurysms,fundus hemorrhage,as well as those who needed retinal laser photocoagnlation and lowering in visual acuity in intensive-therapy group was lower than that in the standard-therapy group (3 vs.14,1 vs.7,2 vs.4,3 vs.11,respectively ;9 vs.36,totally,x2 =4.719,P ＜ 0.05).During the first post-trial monitoring,there was no difference in median HbAlc level between intensive-therapy group and standard-therapy group [(7.2 ± 1.1) ％ vs.(7.3 ± 1.3) ％,t =0.25,P =0.806],which was sustained in the following years.In the trail,no new case of fundus hemorrhage or diabetic associated blindness occurred in intensive-therapy group during the five-year period of therapy.Number of new episodes of micro-aneurysm,macula edema were less in intensive-therapy group than that in standard-therapy group,number of new episodes of lowering in visual acuity,and those who needed retinal laser photocoagnlation,were significantly less in intensive-therapy group than that in standardtherapy group(15 vs.25,4 vs.23,Z =-4.459,P ＜ 0.05) during five-year follow-up.Conclusions The benefit of reduced incidence of diabetic retinopathy in intensive glucose can be maintained because of the legacy effect.

Objective To assess the differential diagnosis and characteristics of renal benign and malignant tumors with three-dimensional contrast-enhanced ultrasound (3D-CEUS). Methods Totally 68 patients with renal tumors were examined by conventional ultrasound and two-dimensional contrastenhanced ultrasound(2D-CEUS). 3D imaging was reconstructed from 2D imaging, the differential diagnosis of renal tumors with 3D-CEUS was analyzed by comparing with 2D-CEUS. All patients with renal tumors were proved by operational pathology. Results Eighteen patients with renal benign tumors mostly displayed equal or low enhancement, showed "slowly in and slowly out" with 2D-CEUS, while displayed regular peripheral and internal vessels with 3D-CEUS. Fifty patients with renal malignant tumors mostly displayed high enhancement, showed "rapidly in and rapidly out" with 2D-CEUS,displayed winding peripheral vessels and disordered internal vessels with 3D-CEUS. 3D-CEUS may display the vascular characteristics of tumors and showed superior imaging quality to 2D-CEUS ( P ＜ 0.05). Conclusions 3D-CEUS can display the vascular characteristics of tumors and their spatial positions, it plays an important role in differential diagnosis between renal benign and malignant tumors.

Objective To explore the value of contrast-enhanced ultrasound(CEUS) in the diagnosis of hematoma and active hemorrhage of renal trauma. Methods Totally 28 patients with renal trauma were examined by conventional ultrasound and CEUS,respectively,including 24 cases caused after renal biopsy,4closed trauma. The detectability of renal hematoma and active hemorrhage with these two methods were compared. All patients were identified by CT or follow-up studies with ultrasound. Results The detectability of renal hematoma with conventional ultrasound and CEUS were 67. 86% (19/28), 92. 86%(26/28), respectively. There was statistically different for detectability in the diagnosis ( P ＜0.05), and the extent of hematoma was more obvious with CEUS. CEUS diagnosed 7 of 26 were renal hematoma with active hemorrhage,which were difficult to be detected with conventional ultrasound. For renal hematoma,the features of CEUS were no enhancement found in every phase; for renal hematoma with active hemorrhage,the contrast agents overflowed from injured blood vessels and formed irregular remarkable enhanced regions. Conclusions CEUS is useful in diagnosing hematoma and identifying the extent and active hemorrhage of renal trauma,in addition,CEUS is valuable in detecting complications after renal biopsy.

AIM:To investigate the changes of Wnt signaling pathway in catalpol-induced proliferation of rat bone marrow mesenchymal stem cells (BMSCs).METHODS:The BMSCs were isolated from SD rats , purified by differ-ential time adherent method and divided into control group and catalpol (1.0 mg/L) group.Flow cytometry was used to de-tect the proliferation index of BMSCs .The mRNA levels of Wnt3a, Wnt5a, Wnt11 and β-catenin was evaluated by real-time PCR.In addition, the protein expression level of β-catenin was determined by Western blotting .RESULTS:Prolife-ration index was increased from 8.90%±0.46% to 17.93%±1.68% after treatment with catalpol (P<0.01).Com-pared with control group , the mRNA expression of Wnt5a, Wnt11 andβ-catenin was all increased with catalpol treatment . No difference of Wnt3a mRNA expression between control group and catalpol group was observed .Meanwhile, the protein expression of β-catenin was increased in catalpol group compared with control group .CONCLUSION:Catalpol promotes BMSCs going into the cell cycle .Classical and non-classical Wnt signaling pathways are activated in this process .

AIM: To study the effect of Bone-benefiting capsule(BBC)-containing serum on activity and apoptosis of osteoclasts(OC) in osteoblasts(OB) and osteoclasts co-culture system.METHODS:(1) OB were separated from the skull of SD rats aged 1 d and OC from the thighbone and shinbone of SD rats aged 5 d,the plane OB and OC co-culture system was established,in which the cells in each culture system did not intermix,but the culture medium communicated,and BBC-containing serum group and control group were concluded in the experiment.(2) Female SD rats aged 10 months were divided into BBC group and control group to prepare BBC-containing serum and control serum.(3) The TRAP secreted by OC was measured by the diazol method and the absorption of lacuna formed by OC was observed by microscope in OB and OC co-culture system to determine the activity of OC with the effect of BBC-containing serum.(4) The effect of BBC-containing serum on OC apoptosis in OB and OC co-culture system was examined by optical microscopes and fluorescence microscopes.RESULTS: In OB and OC co-culture system,the activity of TRAP secreted by OC of BBC-containing serum group were obviously lower than that of control group in 48 h,72 h and 96 h,the OC surviving numbers were obviously lower than that of control group and the rate of OC apoptosis was markedly higher than that of control group which showed an obvious time-efficiency relationship;the amount of the absorption of lacuna formed by OC in BBC-containing serum group was evidently lower than that of control group(P

This study was aimed to investigate the protective effect of catalpol against high glucose induced injury on vascular endothelial cells (ECV-304). ECV-304 cells were culturedin vitro. MTT method was applied to detect cell viability. NO and LDH level were measured by spectrophotometer method. The effect of catalpol against high glucose induced injury on ECV-304 was observed. The results showed that there were no obvious changes in cell viabilities of ECV-304 cultured in normal concentration of glucose medium among groups with different concentrations of catalpol. Compared with the control group, cell viability was significantly decreased (P< 0.01), the NO secretion was significantly reduced (P< 0.01), the LDH level was increased obviously (P< 0.01) in the high glucose (30 mmol·L-1 glucose) injury model group. The high osmotic (24.5 mmol·L-1 mannitol) group was set up as a positive control. Comparison between the low dose (1 ng·μL-1 catalpol + 30 mmol·L-1 glucose), middle dose (10 ng·μL-1 catalpol+ 30 mmol·L-1 glucose), and high glucose (30 mmol·L-1 glucose) injury model group showed no changes on cell viability and NO (P> 0.05); while the LDH level was reduced (P< 0.01). Comparison between the high dose catalpol (100 ng·μL-1 catalpol + 30 mmol·L-1 glucose) group and the high glucose model group showed that the cell viability was obviously increased (P< 0.01); NO secretion was increased (P< 0.05); and the LDH level was obviously reduced (P< 0.01). It was concluded that catalpol had protective effect against high glucose induced ECV-304 injury through the regulation of NO and LDH level. The effect of high dose was obvious.

Objective To analyze the present situation of the dental instrument cleaning and disinfection in Hainan province and to explore the management mode of dental instrument cleaning and disinfection which is suitable for the provincial situation .Meth-ods By adopting the stratified random sampling method according to the hospital grades ,24 hospitals in Hainan province were per-formed the questionnaire survey on the cleaning mode of oral instruments ,layout of cleaning and disinfection room ,cleaning method and facilities ,protection measures and training of cleaning staff .Results 14 hospitals (58 .33% ) had the sterilization and supply center for conducting the centralized processing on the dental instruments .The tertiary hospitals and the second-grade hospitals had the independent cleaning and disinfection rooms with the rational layout and professional cleaning staff ;the safeguard facilities had the application in place ,the training of the related cleaning and disinfection work and the cleaning process conformed the require-ment of the standards .Among 10 first-grade and below hospitals ,only 1 hospital(10 .00% ) had the rational layout of cleaning and disinfection rooms ;3 hospitals(30 .00% ) had the professional cleaning staff ;the related training of the cleaning staff was not basi-cally carried out and the safeguard was not in place ,most of the cleaning and disinfection instruments and the cleaning process were not in accordance with the requirements .The qualification rates of instruments cleaning and disinfection in different grades of hospi-tals by the ATP bioluminescence assay were 100 .00% ,90 .00% and 80 .00% .Conclusion The existing problems are general and prominent in the hospitals of the first-grade and below .It is suggested that the regionalized disinfection and supply management mode is implemented for maximally realizing the optimized resource configuration in the disinfection and supply center .

Objective To discuss the imaging characteristics of chromophobic cell renal carcinoma (CCRC) and study the features on the contrast-enhanced ultrasound (CEUS). Methods The CEUS features of CCRC in 28 cases identified by pathology were reviewed. The blood supply and enhancement characteristic were observed and analyzed on time intensity curve parameters. Results The 28 cases of CCRC showed poor blood supply in contrast with the renal cortex. The CCRC presented with heterogeneity enhancement, part of the tumor took on a high wash-in and wash-out, and enhanced less intense than the surrounding renal cortex. The actinomorphous strong echo of the tumors might be revealed with CEUS in 15 cases (54%). The time intensity curve analysis demonstrated that the CCRCs' difference of peak intensity and area under the curve were lower than the renal cortex (P＜0.05), but arrival time, time-to-peak and slope of ascending curve were higher than the renal medulla (P＜0.05). Conclusion The actinomorphous enhancement and poor blood supply in the tumor of CEUS could provide diagnostic evidence for CRCC.