"Resolving,espulsing and tonifying" three methods are the general outline of endotherapy in surgery of TCM.The author of this article thinks that pelvic inflammation and appendagitis can be classified in internal ulcer and vusceral carbuncle,and it can be treated by "Resolving,espulsing and tonifying" three methods.As the general programme of treating female inflammatory aphoria in different periods,the three methods can avoid exfetatio and promote the probability of pregnancy.Resolving method is applied in acute period of inflammatory aphoria,active period of chronic inflammatory and chronic pelvic inflammation when abdominal mass has formed but vital qi is not deficient.The pathologic character in this period is sufficiency of vital qi and excessive of pathogenic qi.Expulsing method is applied in inflammatory aphoria patients whose pathogenic has not been cleared but the vital qi has been harmed,at this time damp heat pathogen poison remain in uterus meridians and the nvital qi has no power to resist pathogen,and whose corporeity is always week and infects pathogen poison which results in the course of diseases being repeated and prolonged.Tonifying method is applied after the resolving and espulsing methods have been used.In this period,damp heat stasis have been cleared,inflammation and inflammatory matters have been absorbed,disease has been improved and patient has had condition to be pregnant.Tonifying method can promote human generative function and improve the possibility of conception.

BACKGROUND:Preliminary experiments have reported the influence of serum and nerve growth factor on olfactory ensheathing cells proliferation in vitro, but there are less studies concerning choice of serum concentration and growth time for in vitro culture of olfactory ensheathing cells.
OBJECTIVE:To find out the influence of different blood serum concentration and growth time on olfactory ensheathing cells from the olfactory mucosa of adult rats based on the growth curve of olfactory ensheathing cells.
METHODS:The olfactory ensheathing cells from the olfactory mucosa of adult rats were separated, culture and identified in vitro. Sulforhodamine B and microplate reader were employed to measure absorbance values and plot growth curve of olfactory ensheathing cells.
RESULTS AND CONCLUSION:When cultured for the same time in blood serum of different concentrations, absorbance values, especial y in the groups 10%, 20%, 30%, 40%, tended to increase with time except the 0%group. When cultured in the same serum for different time, absorbance values increased within the first 9 days, then promoted rapidly in the groups 10%, 20%, 30%, 40%at 13 days, entered the plateau phase at 19 days, and decreased at 23 days;meanwhile, in the other groups (50%, 60%, 70%, 80%, 90%) the absorbance values peaked at the 13th day and then decreased gradual y. These findings indicate that different serum concentrations and different growth time in vitro affect cellgrowth and survival of olfactory ensheathing cells significantly, which should be ful y considered when cells are cultured in an in vitro condition.

Objective To investigate effects of diabetes mellitus (DM) on pharmacokinetics (PK) of cephradine (CED).Methods DM model was induced by iv.alloxan 60 mg·kg-1.A reversed phase HPLC internal standard method was developed for measurement of CED plasma concentration.After blood collection,rats were sacrificed to collect kidneys for calculating kidney index(KW/BW).DM and normal control (CTL) rats were randomly assigned to receive iv.or ig.CED at a dose of 180 or 90 mg·kg-1.The 3p97 program was used to calculate PK parameters.Results The developed HPLC method was validated to have high specificity,precision,recovery and good storage stability,and met requirements for PK study of CED.The CED in rats of both DM and CTL groups showed the iv.two-compartment PK and ig.one-compartment PK and followed the first-order kinetics.Following iv.dosing,a remarkably decreased t1/2β and MRT,increased CLt were evident in DM group as compared with CTL group (P ＜ 0.05).After ig dosing,a significant decrease in t1/2k and t a remarkable increase in CLt and Cm=were observed for DM group as compared with CTL group (P ＜ 0.05).The DM rats showed a trend of decreased t1/2ka vs CTL rats.There was no significant difference in the oral bioavailability between the two groups (P ＞ 0.05).KW and KW/BW in DM group were increased remarkably compared with CTL group (P ＜ 0.05).Conclusion The DM vs CTL results in faster absorption and elimination of CED in rats,but does not have significantly affect in oral bioavailability.The compensatory hypertrophy and hyperfunction of early-stage diabetic kidneys may constitute one of causes of quick elimination of CED in rats with DM.

Objective To investigate the risk factors for coronary artery lesions (CALs) in children with Kawasaki disease (KD) in Lanzhou. Methods One hundred and seventy-four children with diagnosed KD were divided into CAL group and non-CAL group based on the existence of concurrent CALs. The age, gender, fever duration, intravenous immunoglobulin (IVIG) start time, IVIG dose, C-reactive protein (CRP), serum albumin, erythrocyte sedimentation rate (ESR), platelet (PLT), red blood cell count (RBC), hemoglobin and so on were compared. Results Among the 174 children, 46 children (26.44%) were complicated by CALs and 128 children were not. The differences of average fever duration, IVIG starting time, IVIG dose, PLT, CRP, ESR and RBC were statistically signiifcant (P<0.05). Conclusions When KD children has the fever durations>10 d, start of IVIG af-ter 10 days of fever, increase of PLT, CRP and ESR and decrease of RBC, clinicians should be alert to the risk of concurrent CAL.

Objective:To investigate the effect and mechanism of transcription factor Foxp 3 on the proliferation and cell cycle of human lung adenocarcinoma cell line A 549.Methods:We knocked down the expression of Foxp 3 using siRNA.Foxp3 inhibition was detected by RT-PCR.Cell proliferation was detected by MTT.Cell cycle of A549 cells were detected by flow cytometry after the transfection of siRNA.Cell cycle-related checkpoint genes were filtered by RT-PCR.The regulation of Foxp3 on cell cycle-related checkpoint genes were detected by immunofluorescence and dual -luciferase reporter assay system.Results: The proliferation of A549 cells were inhibited after silencing Foxp3,and A549 cells were arrested in G0/G1 cycle.G1/S cycle checkpoint gene CCND1 was down regulated.Mechanism research show that Foxp 3 can regulate the expression of CCND 1 directly.Conclusion: Foxp3 can promote the proliferation of A549 cell line by up regulating G 1/S cycle checkpoint gene CCND 1.This provides a new target for the therapeutic targets of lung adenocarcinoma.

Objective To investigate the possibility and mechanism of ~ 125 I in treatment of glioma. Methods SHG-44 glioma cells were cultivated in vitro, the inhibitory effect of ~ 125 I on SHG-44 cell proliferation was determined by MTT method. The stereotactic method was used to establish the rat intracranial glioma model. The MRI was performed at 1st week after implantation and ~ 125 I was implanted in the glioma area, the MRI was performed to measure the diameter of tumor 2 weeks after implantation. The rats were killed after 2 weeks ,PCNA gene experession was determined with immunohistological method both in control and experiment group.Results one week after implantation the glioma grew,the results of MTT method showed the growth of the SHG-44 was inhibited, ~ 125 I inhibited the expression of PCNA gene and enlonged the rat survival period. Conclusion ~ 125 I can inhibit the growth of glioma ,the mechanism may be concerned with its inhibitory effect on PCNA gene expression.

OBJECTIVE:To observe the effectiveness and safety of azelastine combined with fluticasone in the treatment of allergic rhinitis(AR). METHODS:A total of 135 AR patients were selected from 3 hospitals during Jan. 2015-Jul. 2016, and divided into azelastine group,fluticasone group and drug combination group according to random number table,with 45 cases in each group. On the basis of decongestant Oxymetazoline hydrochloride spray,azelastine group was given Azelastine hydrochloride nasal spray,1 spray each nostril,morning and night;fluticasone group was given Fluticasone propionate nasal spray,1 spray each nostril,morning and night;drug combination was given Azelastine hydrochloride nasal spray combined with Fluticasone propionate nasal spray,with the same usage and dose. Three groups were treated for consecutive 30 d. Total nasal symptom score (TNSS) and total ocular symptom score (TOSS) decline indexes,nasal minimal cross-sectional area (NMCA),total nasal resistance(TNR)and the occurrence of ADR were compared among 3 groups. RESULTS:Some pa-tients were withdrawal from the trial;finally,127 patients were included in full analysis set,and 130 patients were included in safety set. After treatment,TNSS decline index of drug combination group was significantly higher than azelastine group and fluticasone group,with statistical significance(P<0.05). There was no statistical significance in TNSS decline indexes be-tween azelastine group and fluticasone group,as well as TOSS decline indexes among 3 groups(P>0.05). Before treatment, 3 groups suffered from nasal ventilation disorder to different extents,but there was no statistical significance in TNR or NMCA among 3 groups(P>0.05). After treatment,NMCA and TNR at 75,150 Pa of 3 groups were significantly lower than before,and TNR of drug combination group was significantly lower than azelastine group and fluticasone group,with statisti-cal significance(P<0.05). There was no statistical significance in NMCA among 3 groups(P>0.05). There was no statistical significance in the incidence of ADR among 3 groups(P>0.05). CONCLUSIONS:Both azelastine and fluticasone can effec-tively relieve nasal symptom as well as improve nasal ventilation disorder in AR patients;therapeutic efficacy of drug combi-nation is better than single drug,and it doesn't increase the incidence of ADR.

AIM: To assess the effect of raloxifene, a selective estrogen modulator, on the transcriptional activity of estrogen responsive element (ERE) in vascular endothelial cells. METHODS: Vascular endothelial cells were transfected with ERE-TK-Luc reporter plasmid and PRL-SV40 control plasmid via FuGENE 6. The activities of firefly luciferase and renilla luciferase were measured with dual-luciferase reporter assay system. The transcriptional activity in transfected cells treated with raloxifene was compared with that in untreated cells. Furthermore, raloxifene was used to cotreat the cells with estradiol (E_2) and the influence of raloxifene to the effect of E_2 was assessed. RESULTS: Compared to untreated cells, the luciferase activity in cells treated with raloxifene decreased and showed significance at concentration of 10~(-7)mol/L (P

BACKGROUND:Exogenous neurotrophic factors or chemical induction can induce rat bone marrow mesenchymal stem cells to differentiate into neuron-like cells. However, exogenous inductors exert a short inducible action, and their chemical substances inevitably have a negative impact on cellviability to limit the application prospects of bone marrow mesenchymal stem cells to a certain extent.
OBJECTIVE:To investigate the effect of glial cellline-derived neurotrophic factor, green fluorescent protein gene transfection by adenovirus vector on biological characteristics of rat bone marrow mesenchymal stem cells, to observe the expression of glial cellline-derived neurotrophic factor and green fluorescent protein and the role of nutrition on bone marrow mesenchymal stem cells, and to explore the ability to differentiate into neuron-like cells induced by glial cellline-derived neurotrophic factor.
METHODS:The bone marrow mesenchymal stem cells at passage 3 were transfected by recombinant adenovirus (Multiplicity of infection=10, 50, 80, 100, 150, 200). The experiment had two groups according to target genes:bone marrow mesenchymal stem cells were transfected by Ad-GDNF-GFP in transfection group, and bone marrow mesenchymal stem cells were not transfected in control group. The expression of green fluorescent protein was detected by inverted fluorescence microscope. Transfection efficiency was calculated by flow cytometry. cells viability and the morphological changes of cells were compared respectively by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and inverted fluorescence microscope between the two groups. On days 5 and 10 after transfection, the expression of glial cel-derived neurotrophic factor mRNA was detected by PCR. On day 5, the expression of neuron-specific enolase was determined by immunofluorescence examination. On day 10, the expression of microtubule-associated protein 2 was identified.
RESULTS AND CONCLUSION:By the end of 12 hours after transfection, the green fluorescent protein expressed in cells, and the fluorescence intensity gradual y increased with time. When the multiplicity of infection was 100, the fluorescence intensity was strong and stable, and the transfection rate was nealy 90%on day 3 after transfection. cellviability in the transfection group was strengthened after transfection. On day 5 after transfection, bone marrow mesenchymal stem cells expressed neuron-specific enolase, and neuron-like protrusions gradual y extended. On day 10 after transfection, bone marrow mesenchymal stem cells expressed microtubule-associated protein 2 and glial cellline-derived neurotrophic factor mRNA, and exhibited neuron-like morphology and interconnected synpases. The recombinant adenovirus, Ad-GDNF-GFP, can highly transfect bone marrow mesenchymal stem cells when the multiplicity of infection is 100, and glial cellline-derived neurotrophic factor can promote the proliferation of bone marrow mesenchymal stem cells and induce bone marrow mesenchymal stem cells to differentiate into neuron-like cells.

BACKGROUND:Scaffolds made of chitosan and its derivatives play an important role in cellmigration and axonal regeneration. Chitosan and its derivatives have good histocompatibility, which is easy to make stem cells grow on the surface, thereby having a more broad application prospect in the nerve tissue engineering.
OBJECTIVE:To fabricate a thermosensitive hydrogel scaffold using chitosan/hydroxypropyltrimethyl ammonium chloride chitosan/sodium glycerophosphate (CS/HACC/GP), which is suitable for cellgrowth, and then, to observe the growth and proliferation of bone marrow mesenchymal stem cells on the scaffold.
METHODS:Chitosan was modified using quaternary ammonium salt and confirmed by Fourier transform infrared spectroscopy. The chitosan and quaternary ammonium salt of chitosan was mixed at a ratio of 8:1 to successful y prepare stable CS/HACC/GP thermosensitive hydrogel scaffold. Then, the gel ing was observed, and biosafety test was conducted.
RESULTS AND CONCLUSION:Fourier transform infrared spectroscopy showed the characteristic peak of quaternary ammonium groups. Cytotoxicity test showed that rat bone marrow mesenchymal stem cells cultured in hydrogel extracts had no toxicity. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test showed that hydrogel extracts exerted no significant effect on the increase in body weight, and the biological safety of the scaffold was good. Under the scanning electron microscopy, bone marrow-derived mesenchymal stem cells grew and proliferated normal y in the scaffold. The results confirmed that the CS/HACC/GP thermosensitive hydrogel scaffold was successful y prepared in the experiment, which is suitable for the growth and proliferation of bone marrow mesenchymal stem cells.