0.01). Conclusion: Occlusal reduction might diminish the rate of postoperative pain following endodontic instrumentation.

BACKGROUND:The growth of mesenchymal stem cells in vitro in different conditioned media is different evidently. So it is necessary to choose a more suitable medium. OBJECTIVE:To contrast and observe the proliferation of human umbilical cord mesenchymal stem cells in three kinds of media and to check the immunophenotype and differentiation ability of mesenchymal stem cells. METHODS:Human umbilical cord mesenchymal stem cells were col ected by explant method in sterile conditions. After subculturing by T75 incubation bottles, the third generation of mesenchymal stem cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium containing 5%fetal bovine serum, Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum and Mesen PRO RSTM medium. After 1, 3, 5, 7 days of culture, the cells were counted to draw a growth curve. Immunophenotype of the third generation of umbilical cord mesenchymal stem cells were determined by flow cytometry and the ability of osteogenic and adipogenic differentiation was also detected. RESULTS AND CONCLUSION:The third generation of cells cultured highly expressed CD44, CD73, CD90, CD105, but did not express CD29, CD31, CD34, HLA-DR. The oil red O staining showed a lot of little red lipid drops after adipogenic induction;alizarin red staining showed osteoblast-like cells after osteogenic induction, indicating umbilical cord mesenchymal stem cells in vitro have the potential of multi-directional differentiation. After observation and counting, the colony and shape of cells cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum were superior to those cultured in the other two media. Therefore, it is concluded that Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum is preferred for cellsubculture.

Objective To investigate the effect of high-mobility group box protein1 (HMGB1) on the expression of TNF-αand its mechanism in 16HBE in vitro. Methods groups with different HMGB1 (0, 100, 500, 2 000 ng/mL) concentration was set; RAGE antagonizing groups were as control, HMGB1-2000ng, anti-RAGE and anti-RAGE+HMGB1. The changes of TNF-αmRNA and secretion were determined by quantitative PCR and ELISA. RAGE protein level was measured by western blotting. Results HMGB1 intervention and TNF-α expression of 16HBE presented a positive dose-dependent relationship. Thechanges of RAGE was HMGB1positively concentration dependent. In comparison with HMGB1 2 000 ng/mL group, anti-RAGE+HMGB1showed a remarkable reduction of TNF-α secretion. Conclusion In vitro, HMGB1 increases TNF-α expression in 16HBE with a dose-dependent manner through RAGE.

Objective To establish a real‐time quantitative PCR method for the detection of cytokine receptor‐like factor 2 (CRLF2) expression .Methods Specific primers amplification target gene CRLF2 and housekeeping genes ABL were designed ,the purified PCR products were performed the TA cloning .After bacterial colony PCR screening and sequencing ,then the recombinant plasmids DNA was extracted and measured by using UV spectrophotometer and converted to copies/mL concentration .Finally it was diluted for preparing the plasmid standard substance ,then the standard curve was drawn for observing the sensitivity and linear rang ,meanwhile the stability of the plasmid DNA was evaluated .This method was initially applied to detect the CRLF2 level of bone marrow mononuclear cells in 10 cases of healthy children and 10 cases of newly diagnosed acute lymphoblastic leukemia (ALL) .Results CRLF2 PCR product had a single specific melting curve;the linear detection range of the standard substance was 103 - 108 copies /ml;the plasmid standard substance by freeze‐thawing for 3 times remained stable;the CRLF2 level of clinical sample was within the linear detection range of standard substance .Conclusion The real‐time quantitative PCR method for CRLF2 established by our laboratory has good specificity ,linearity range and stability ,which can be applied to the quantitative detection of CRLF2 gene in clinical ALL children .

Objective To establish the murine animal model for acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation (HSCT). Methods T cell-depleted (TCD) BM cells from allogeneic C57BL/6 mice were harvested and prepared from the marrow of the femurs and tibias. Age of 4 to 6 weeks’ BALB/c mice received 900 cGy total body irradiation ( TBI) from a 137Cs source. Two to four hours later,the mice were injected intravenously ( i. v. ) 5 × 106 TCD-BM cells with 1. 25 × 106 or 2. 5 × 106 or 5 × 106 splenic cells from allogeneic C57BL/6 mice,and respectively divided them into experimental group 1,experimental group 2 and experimental group 3. TCD-BM only was used as the negative control group of GVHD. Ten mice were used for each group. The establishment of aGVHD model was evaluated with a clinical GVHD scoring system,survival rate and target-organ damage. Results On 7 to 14 days after transplanta-tion,the typical clinical GVHD manifestation of severe diarrhea,hogback,activity reduced and ruffling were observed in experimental group 2. Furthermore,the aGVHD target organs of colon,lung and liver were harvested and made histological paraffin sections,then the obviously path-ological tissue damages of GVHD were detected under microscope. And the survival rate was lowed down to 0 on 45 days after transplantation in experimental group 2. On the contrary,no obviously clinical manifestation of aGVHD were observed in the control group,experimental group 1 and group 3. On 60 days later, the survival rate was 80% in experimental group 1 and 100% in control group. However,no mice was sur-vived on 10 days later in group 3. Conclusion BALB/c mice aGVHD model after allogeneic HSCT is successfully established by injecting i. v. 5 × 106 TCD-BM cells with 2. 5 × 106 splenic cells from allogeneic C57BL/6 mice.

Photothermal therapy (PTT) has attracted significant attention due to minimal side effects and high treatment specificity. However, it often requires very high temperature to achieve complete tumor ablation under a single PTT. Such high temperature brings obvious thermal damage and inflammatory response to the body, affecting the therapeutic effect. In recent years, nitric oxide (NO) has been used to significantly inhibit tumor growth and enhance the sensitivity of tumor cells of temperature and drugs, thus enhancing the therapeutic effect. However, compounds as NO donors often have some disadvantages such as poor biocompatibility and untargeted delivery, etc., therefore, this medical application based on NO therapy is limited. In conclusion, the organic combination of NO donors and photothermal agents (PTAs) is expected to overcome the shortcomings of single therapy and achieve the antitumor effect of "1 + 1 > 2". In view of the rapid development of NO combining with PTT in tumor therapy, this review firstly introduces the antitumor mechanisms of different types of NO donors. Then the treatment strategy based on NO combined with PTT is discussed. Finally, the prospects and challenges of this combination therapy strategy in the clinical treatment of cancer are discussed.

Objective To observe the variation of enzymatic activity and areas and bulk of focus of heart injuries by using controllable catheter to ablate epicardial tmsue of rabbits and focus underneath atrioventrieular ring narcosis with 20% urethane(4 ml/kg)and divided into three groups.Each group included 7 rabbits.Anterior wallepieardium of left ventricle was ablated thirty seconds in each group(10,20 and 30 W)with self-made ablationspheroid microwave antenna,refilling with high pressure normal saline at same time.Then all of the rabbits were sacrificed respectively and their ventricular myocardium were taken out to undergo immunohistochemistry in order to display suceinate dehydrogenase(SDH).Also amplitude Wag measured in order to calculate areas of heart injuries.(8F)wag delivered to the pre-selected sites around atrioventricular ring of thirty-two healthy dogs,which had beenin intravenous narcosis with pentobarbital sodium(30 mg/kg).The dogs were divided into four groups(40,50,60 and 80 w) and two time points(60 and 120 s),by the combined method of X-ray and endocardial electrocardiograph,the microwave antenna could be confirmed to be located at the accurate position between anterior and posterior wall close to septum of left/right ventricle.After ventricular myocardium had been taken out,amplitude were measuredin order to calculate bulk of heart injuries by 1/6×3.14 x long×wide×deep.In addition.the histological changesand transmural injury were examined by optic microscope.Results In each group,the centre of injuries wagenzyme deficiency locus.The diameter and areag of heart injuries enlarged significantly(3.99.±0.41),(5.20±0.25),(6.31±0.37)mm and(12.53±2.56),(21.19±3.14),(30.96±3.76)mm2 with the increased microwave power level(10、20、30 W).Group comparison had statisficM significance(F＝76.8,58.5;P＜0.01 or ＜0.05).A total of 116points were ablated.The myocardial lesion showed ellipse in shape,and continuous symmetrical coagulationnecrosis under microscopic examination.There was a clear demarcated line around tlle myocardial tissue and fewparietal thmmbus.There were 16 transmura]injuries and five-with lung damage.The bulk of lesion aroundatrioventrieular ring hag been significantly enlarged(46.7±2.5),(51.1±2.7),(133.2±3.4),(141.8±3.9),(248.5±6.2),(260.3±6.5),(313.7±9.5),(327.4±10.5)with the increased microwave power level(40,50,60and 80 W)and/or distance of microwave ablation(60 and 120 s).Groups comparison had statistical significance(F＝31.16,27.85;all P＜0.01).In each time point,the lesion bulks had conspicuous distinction of statistics.In the same microwave power,the time wag longer,the bulk was larger(P＜0.01).Conclusions The more the microwave power level and time,the severe the heart injuries is.It is possible to use the microwave energy to ablate the deep focus under endocardium around atrioventricular ring.

AIM: To investigate the alterations of tear film after sutureless large incision manual cataract extraction (SLIMCE). METHODS: Sixty-eight SLIMCE operation eyes were studied with slit-limp microscope, break- up time (BUT), SchirmmerⅠtest (SⅠt),and fluorescence(FL) to observe the alterations of tear film at different time points in postoperation. Impression cytology and microphoto-analyses technique were also applied to observe the goblet cells at different time points postoperation(7,14,30,60,90 days). RESULTS: Subjective complaint of dry eye within 90 days after the operations were significantly increased compare with preoperations(5-27,23,19,16,13; 2-16,14,8,6,3). The schirmmer Ⅰ test were greatly increased in 14 days postoperation(10.1±4.5;15.0±4.7,13.8±5.7),the mean scores of fluorescence increased (0-17,9,5;0-8,3,1) and the mean break-up time decreased in 30 days post-operation(10.3±2.2;5.5±2.3,7.0±2.4,7.9±2.2) (P<0.05). CONCLUSION: SLIMCE operation have effect on the stability of tear film.

OBJECTIVETo observe the effect of cold or hot properties of traditional Chinese medicines (TCM) on biological effect indexes, and analyze the contribution of variables on cold or hot properties, in order to preliminarily establish the discrimination mode for the biological effects of cold or hot properties.

METHODRats were randomly divided into the blank control group, cold TCM groups (Coptidis Rhizoma, Scutellariae Radix, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix and Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex and Evodiae Fructus), and orally administered with 10 mL x kg(-1) of corresponding TCM water decoctions for 30 d, twice a day. Altogether 53 biological effect indexes correlated to cold or hot properties of traditional Chinese medicines were founded by searching literatures. The data warehouse were established by using data-mining software Clementine12.0. Data of the blank control group, cold TCM groups (Coptidis Rhizoma, Phellodendri Cortex, Gardeniae Fructus, Sophorae Flavescentis Radix, Gentianae Radix) and hot TCM groups (Aconiti Lateralis Preparata Radix, Zingiberis Rhizoma, Alpiniae Officinarum Rhizoma, Zanthoxyli Pericarpium, Cinnamomi Cortex) were selected into a training set. C5.0 algorithm and C&R classification and regression algorithm were adopted to define the importance of variable, create the decision trees, and test hot or cold properties of Evodiae Fructus and Scutellariae Radix.

RESULTAccording to C&R classification and regression algorithm, SDH activity of livers was the most important hot or cold property, with the significance closed to 30%. It was followed by triglyceride, liver Na' -K' -ATPase enzyme, muscle glycogen and platelet distribution width, with the accuracy up to 97.39% in models. C5.0 algorithm showed that liver SDH activity was the most important hot or cold property, with the significance closed to 40%. It was followed by triglyceride, GOT, muscle glycogen and liver Na(+)-K(+)-ATPase enzyme, with the accuracy up to 98.26% in models. The possibilities that Evodiae Fructus is in hot property and Scutellariae Radix is in cold property were 100. 00% and 77.78% by using both C&R classification and regression algorithm and C5.0 algorithm.

CONCLUSIONThe SDH activity of liver is the most important biological effect index to distinguish cold and hot properties of TCMs. The discrimination pathway or mode of cold and hot properties is closely related to energy metabolism.

Algorithms ; Animals ; Drugs, Chinese Herbal ; classification ; pharmacology ; Fruit ; chemistry ; Liver ; drug effects ; metabolism ; Liver Glycogen ; metabolism ; Male ; Medicine, Chinese Traditional ; methods ; Outcome Assessment (Health Care) ; methods ; Phytotherapy ; classification ; methods ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; classification ; Random Allocation ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Succinate Dehydrogenase ; metabolism ; Triglycerides ; metabolism

Objective To analyze the factors that influence the disease-free survival time after radical prostatectomy for prostate adenocarcinoma and to study the expression and significance of Bcl-2,C-erbB-2,Ki-67 in prostate adenocarcinoma. Methods A total of 51 cases of prostate adenocarcinoma undergoing radical prostatectomy were reviewed.Cox proportional hazard model was used to investigate the impact and co-impacts of the factors,which included age, preoperative PSA level,Gleason scores of biopsies and operative specimens,EPE,MR,SVI,LN,PAT,PRT and having testes or not during follow-up.Immunohistochemical staining was used in biopsies and resected specimens to evaluate the positivity of Bcl-2,C-erbB-2,Ki-67 in 15 of the 51 cases. Results In stepwise Cox proportional hazards model, the prognostic factors for recurrence were age(P=0.011),maximal androgen blockade(P=0.017), positive margin of resection(P=0.000),postoperative Gleason score(P=0.002),having testes or not after operation(P=0.040).The expressions of 3 types of proteins showed negative correlation with the prognosis of prostate carcinoma. Conclusions Among clinicopathologic factors,positive resected margin is the most powerful prognostic factor.Patients will benefit from maximal androgen blockage before radical prostatectomy.Disease-free survival after radical prostatectomy is not influenced by not having pelvic lymphadenectomy after finding no enlarged lymph nodes intraoperatively.Not preserving testes after prostatectomy is a favorable factor.The prognosis of prostate carcinoma can be predicted with the expressions of Bcl-2,C-erbB-2 and Ki-67 in biopsies or resected specimens.