Objective: To find out the parameters of drop pill's process in proportion to matrix, refrigerant and addition, in relationship with the height and rotational speed of freeze-pillar and drop pills. Methods: Using parallel experimentation for the choice of matrix and refrigerant; mode and proportion of adding drug. Using orthogonal experimentation for the choice of height and rotate speed about freeze-pillar, dropping speed. With moulding probability of drop pills as the evaluation quota to decide the best dropping condition. Results: PEG 4000 is matrix. Methyl-siliconoil: Liquid-paraffin (3∶1) is refrigerant. Internal and external diameter of burette are equal to 4.5mm and 5.8mm. Drug is dropped into the refrigerant of 10～15 ?C by 50 to 60 dropping per minute (freeze-pillar height 120cm, rotate speed is 20 round per minute.) Moulding probability is of the highest one. Conclusion: The moulding technics of drop pills with high finished product rate and good quality.

24 cases of renal failure induced by chemotherapy were treated with Wuling powder plus additives.Theresult of clinical amelioration was 87.5%.with a to-tal effective yielded marked actions of promoting ap-petite in animals,lowering serum urea nitrogen,cre-atinine,2 microglobulin.It also decreased the inhibi-tion of PDD on the enzyme activity of renal Na~+—K~+ATP.Microscopic exam revealed the pathological le-sion of kidney to be milder in the tested group.

Objective To investigate the influence of isoquercitrin on the inflammatory factors in LPS-induced RAW264.7 cells.Methods MTT method was used to detect inhibition ratio of RAW264.7 cells induced by isoquercitrin.The level of TNF-α in culture medium was measured by ELISA.Nitric oxide (NO) was detected by Nitrate Assay Kit.Western blotting was used to investigate the influence on the productions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).Results The half inhibitory concentration (IC50) of isoquercitrin was 65.73 μmol·L-1.LPS had no inhibitory effect on the cells.Compared with LPS group,the level of TNF-α was decreased to 74.80% and 60.57% in isoquercitrin (20,10 μmol·L-1) groups in a dose-dependent manner.The results measured by Nitrate Assay Kit revealed that isoquercitrin (20,10 μmol·L-1) could suppress production of NO,the level of NO decreased to 79.34% and 68.81%(P<0.05).The Western blotting results showed that isoquercitrin (20,15,10 μmol·L-1) inhibited the productions of iNOS and COX-2 (P<0.05).Conclusion Isoquercitrin has anti-inflammatory effects by inhibiting the productions of TNF-α,NO,iNOS and COX-2,and the most effective dose for the inhibition is 10 μmol·L-1.

Aim To study the inhibitory effect of isoquercitrin on Raf/MEK/ERK signaling pathway in HepG2 cells.Methods MTT was used to detect the proliferation of human liver cancer HepG2 cells after the treatment of isoquercitrin.The morphology and growth of cells were observed under inverted microscope after the different concentrations of isoquercitrin(0, 40, 80, 160, 320 μmol·L-1) to treat HepG2 cells for 24 and 48 h.Cell cycle was assessed by flow cytometry.Ras, Raf, MEK, ERK expression was assayed by Western blot, and mRNA expression was detected by quantitative fluorescence PCR.Results Isoquercitrin could inhibit the growth of HepG2 cells in a concentration-and time-dependent manner.Typical morphological changes of apoptosis were observed by inverted microscopy after HepG2 cells were treated with different concentrations of of isoquercitrin for 24 h or 48 h.The cell cycle assay showed that with the increasing concentration of isoquerditrin, the number of cells that was arrested in G1 phase gradually increased.Compared with the blank group, the expressions of Ras, Raf, MEK, ERK mRNA were down-regulated, and related proteins expression were also down-regulated(P<0.05), and these results had statistical significance.Conclusion Isoquercitrin can induce the apoptosis of HepG2 cells, which may be related to the Raf/MEK/ERK signaling pathway.

Objective To investigate whetherthe extract of HUANGPI inhibitthe secretion of TNF-αvia TLR4/MyD88/TRAF6 signaling pathway. Methods ELISA assay was performed to determine TNF-α level in cell culture medium. MTT assay was used to detect the effects of the extract of HUANGPI and LPS on the viabilities of RAW 264.7 cells. Proteinexpressions of TLR4 and TRAF6 were detected by Western blotting assay. Results The extract of HUANGPI inhibited the secretion of TNF-αin a dose-dependent manner. Compared to LPS group , were TNF-αwas significantly suppressed in the cells in LPS+MyD88 inhibitor group , LPS+extract group and LPS+extract+MyD88 inhibitor group,with the corresponding reductions of TLR4 and TRAF6 protein expression at74% and21%,70% and27%,44% and8.5%, respectively. Conclusion MYD88-dependent signaling pathway might be involved in the mechanism underlying the effect of the extract of HUANGPI on suppressing LPS-induced inflammation.

Objective To monitor the autophagy in osteosarcoma cells by constructing three rLC3B fusion expression vectors,respectively.Methods Rat LC3B gene sequence was amplified by PCR and cloned into pEGFP-C 1 and pmCherry-C1 to construct the fusion expression vector of pEGFP-rLC3B and pmCherry-rLC3B.Subsequently,the EGFP-rLC3B sequence was obtained by PCR with the pEGFP-rLC3B as a template,and cloned into pmCherry-C 1,so the pmCherry-EGFP-rLC3B fusion expression vector was constructed.Three plasmids were transfected into U-2OS cells,and the starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A1 was used to inhibit autophagy,to verify the above plasmids' function in autophagy detection by laser scanning confocal microscopy.Western blot was used to detect the endogenous LC3B and exogenous EGFPrLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B,and to verify the correct expression of exogenous rLC3B and their function of autophagy detection.Finally,cleaved free EGFP was detected by western blot to evaluate the level of autophagic degradation.Results Three fusion expression vectors were constructed successfully through sequencing and restriction enzyme digestion validation.The starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A 1 was used to inhibit autophagy in transfected U-2OS cells.Clear autophagosomes and autolysosomes were observed by laser scanning confocal microscopy.Endogenous LC3B and exogenous EGFP-rLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B were detected through western blot.Finally,western blot verified that the expression of cleaved free EGFP was significantly up-regulated with the increase of starvation time.12 h group increased 1.05 times than the control group and 24 h group increased 1.56 times,showing that the levels of autophagic degradation increased.Conclusion EGFP-rLC3B can be used to detect autophagosome and evaluate the level of autophagic degradation.mCherry-rLC3B can be used to detect autophagosome and autolysosome,but can't distinguish autophagosome from autolysosome.The pmCherry-EGFP-rLC3B has an advantage in the detection of autophagic flux which can distinguish autophagosome from autolysosome.

Objective To explore oropharyngeal swallowing disorders with videofluoroscopic swallowing study (VFSS). Methods 16 patients with dysphagia accepted VFSS with 10 ml of thin barium meal (50% w/v), thick barium meal (270% w/v), biscuit coated with thick barium meal in single swallow. Their swallowing function was observed on the lateral and anterior/posterior planes, including: symmetry of pyriform sinuses, oral transit time, presence of pharyngeal delay, pharyngeal transit time, oral and pharyngeal residue, and presence of aspiration.Results 5 patients demonstrated oral swallowing disorder. 3 patients demonstrated pharyngeal swallowing disorders, that was pharyngeal delay which caused in aspiration after swallowing. 8 patients demonstrated oropharyngeal swallowing disorders, and 3 of them presented aspiration,2 patients were silent aspirators, 1 was aspiration before and 1 after swallowing. The aspiration time could not be judged from the videofluoroscopy in the other one. For 4 patients with aspiration, 3 were severe, with more than 25% of the bolus aspirated, and 1 aspirated less than 5%. Conclusion VFSS can be helpful to plan individual rehabilitation.

Clinical observation revealed that combined use of Decoction of Radix Astragali seu Hedysari and Poria withlarge dose of Cisplatin.none of the cases showed ab-normality in BUN,Cr.with improvement of biochemi-cal indices,there was also diuretic actions observed.Ahgdraton contrel group revealed worse result.The ex-periment proves that the Decoction is capable of pre-venting and ameliorating the harmful effect on kidneyinduced by Cisplatin in rats.

Objective To evaluate the two-stage repair in hypospadias cripples.Methods 35 patients with hypospadias cripples underwent two-stage repair.At the first stage,a full-thickness graft of skin or buccal mucosa was used for urethral plate reconstruction after the release of chordee.The second stage was performed 6 months after completion of the first stage.Tubularization of the replaced urethral plate was the preferred technique.Results There was no urethra stricture or “preplaced urethral plate ” atrophy.Fistula developed in 2 of 35 cases.Conclusion The twostaged technique was a useful strategy to deal with the myriad abnormalities encountered in crippled hypospadias.This technique could not only create a neourethra successfully,but also give the penis a near-normal shape and appearance.

Objective To retrospectively analyze treatment status quo of giant cell tumor of bone around the knee in several institutes,and to investigate risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes.Methods A total of 222 patients with giant cell tumor of bone around the knee confirmed by pathology,who had undergone surgical treatment in 5 institutes from March 2000 to May 2012,were enrolled in this study.There were 120 males and 102 females,with an average age of 35.5 years.The epidemiology,clinical and radiographic features and risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes were analyzed.Results Intralesional curettage was performed in 128 patients(57.6％),intralesional curettage combined with partial resection in 13 patients (5.8％),and marginal excision in 79 patients (35.5％).A total of 159 patients were followed up.The local recurrence rate was 19.0％ for patients treated with intralesional curettage,8.9％ for marginal excision and 0％ for intralesional curettage combined with partial resection,and the total local recurrence rate was 14.5％(23/159).The mean duration from primary operation to recurrence was 23.9 months.Univariate analysis indicated that surgical manner was the only factor affecting local recurrence.Pathological fracture,Campanacci grades,Enneking system as well as soft tissue mass had a significant influence on the selection of surgical manners.The mean MSTS score after intralesional curettage was obviously higher than that after marginal excision.Conclusion The surgical manner is the only factor affecting local recurrence and limb function.