In order to test the equilibrium solubility of puerarin in different solvents and solubilizer,cilia toxicity and irritation of these excipient, the balance method, toad in the ciliary body toxicity and rat nasal mucosa irritation were used respectively. Results showed that puerarin solubility was 56.44 g x L(-1) in combined solvent of 30% PEG200 and 10% Kolliphor HS 15. With normal saline solution as negative control and sodium deoxycholate as positive control, the effects of 30% PEG200, 30% PEG 400, 10% Kolliphor HS 15 and combination of 30% of PEG200 and 10% Kolliphor HS 15 on toad palate cilium were observed and cilia movement duration was recorded. The results indicated that there was no significant difference in cilia movement duration among 30% PEG200, 10% Kolliphor HS 15 and normal saline group. The rats long-term nasal mucous membrane irritation of 30% PEG 400, 10% Kolliphor HS 15, which had no cilia toxicity, was studied, with normal saline solution as negative control. There were no significant difference revealed on rat nasal mucosa epithelial thickness among 30% PEG 400, 10% Kolliphor HS 15 and normal saline. Above researches showed 30% PEG 400, 10% Kolliphor HS 15 was ideal for solubility of puerarin nasal drops and showed a lower cilia toxicity and irritation, and can be used as the solvent and solubilizer of puerarin nasal drops.
Administration, Intranasal ; methods ; Animals ; Anura ; Cilia ; chemistry ; Female ; Isoflavones ; chemistry ; Male ; Nasal Mucosa ; Polyethylene Glycols ; chemistry ; Rats ; Rats, Sprague-Dawley ; Solubility ; Solvents ; chemistry

The stability of melittin in different solvents (water, deoxygenated water, physiological saline, PBS, 50% ethanol, ethanol, glycerol)was studied and the results showed that the stability of melittin is not influenced by light, temperature and pH in 50% ethanol, which melittin can be completed dissolved when compared with ethanol and glycerol, in such, 50% ethanol was chosen as solvent storage when measured content of melittin. Then the effect of different concentrations of PBS, the pH of PBS and rat skin ho- mogenates were tested, and the results showed that melittin was degraded rapidly at low concentration solution and low ionic strength. Increasing pH of PBS and rat skin homogenate can accelerate the degradation of melittin. These researches provide an experimental ba- sis for further study of melittin.
Animals ; Drug Stability ; Ethanol ; chemistry ; Hydrogen-Ion Concentration ; Melitten ; chemistry ; Rats ; Skin ; drug effects ; Solvents ; chemistry ; Temperature

(3S)-Linalool synthase (LIS) is a key enzyme involved in the monoterpene biosynthetic pathway. Based on our previous transcriptome study, the expression level of LIS gene was exceedingly related to glycyrrhizic acid (GA) biosynthesis. Therefore, we used hairy root culturing to further investigate the effect of LIS on the GA biosynthesis. A LIS gene (GenBank accession number: MZ169552) was cloned from Glycyrrhiza uralensis. The plant binary overexpression vector pCA-LIS was constructed by gene fusion. G. uralensis hairy roots overexpressing LIS were induced by the Agrobacterium rhizogenes ATCC15834. The expression levels of LIS were analyzed by real-time quantitative PCR (RT-qPCR) and the contents of GA in hairy root lines were determined by UPLC. It was found that in the hairy root lines overexpressing LIS, the expression levels of LIS were significantly higher than that in the wild type, while the contents of GA were remarkably lower than those in the wild type and negative control. These findings indicate that the expression level of LIS is negatively correlated with the accumulation of GA. In this study, LIS was cloned from G. uralensis for the first time and the negative regulatory effect of LIS on GA biosynthesis was confirmed by reverse genetics. This work provides support for further improvement of the molecular regulatory network of GA biosynthesis in G. uralensis.

In order to investigate the characteristics of transdermal delivery of ferulic acid under the treated of microneedle arrays and the influence on permeability of rat skin capillaries, improved Franz-cells were used in the transdermal delivery experiment with the rat skin of abdominal wall and the length of microneedle arrays, different insertion forces, retention time were studied in the influence of characteristics of transdermal delivery of FA. The amount of FA was determined by HPLC system. Intravenous injection Evans blue and FA was added after microneedle arrays treated. Established inflammation model was built by daubing dimethylbenzene. The amount of Evans blue in the rat skin was read at 590 nm wavelength with a Multiskan Go microplate reader. Compared with passive diffusion group the skin pretreated with microneedle arrays had a remarkable enhancement of FA transport (P <0.01). The accumulation of FA increased with the enhancement of insertion force as to as the increase of retention time. Microneedle arrays with different length had a remarkable enhancement of FA transport, but was not related to the increase of the length. The research of FA on the reduce of permeability of rat skin capillaries indicated that the skin pretreated with microneedle arrays could reduce the content of Evans blue in the skins of rat significantly compared with the untreated group. The permeation rate of ferulic acid transdermal delivery had remarkable increase under the treated of microneedle arrays and the length of microneedle arrays ,the retention time so as to the insertion force were important to the transdermal delivery of ferulic acid.
Administration, Cutaneous ; Animals ; Coumaric Acids ; administration & dosage ; pharmacokinetics ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Skin Absorption

Objective To analyze the efficacy,late complications and prognostic factors of post-op-erative radiotherapy for cervical cancer. Methods From Nov. 1999 to Feb. 2005,114 patients with cervi-cal cancer received adjuvant pelvic radiotherapy after radical hysterectomy and pelvic lymphadenectomy. The median age was 42.5 (24 to 72) years old. According to the FIGO staging system,6,51,18,26 and 13 pa-tients had stage ⅠA, Ⅰb1, Ⅰb2, ⅡA and ⅡBdisease. The pathological type was squamous cell carcinoma,ade-nocarcinoma, squamous-adenocarcinoma and undifferentiated carcinoma in 92,19,2 and 1 patients, respec-tively. The whole-pelvic external beam irradiation of 50 Gy (40 to 60 Gy) was given with 6 MV or 15 MV X-ray beams using four-field box technique. Eighty-one patients received intravaginal brachytherapy of 16 Gy (4 -30 Gy in 1 -6 fractions) 4 weeks after the beginning of radiotherapy, with the referrence point being at 0.5 cm under the vaginal mucosa. Eighty-seven patients received preoperative and/or concurrent chemother-apy. The survival and independent prognostic factors were analyzed by Log-rank method and Cox model. Results The median follow-up time was 26.0 (5 - 75) months. The overall survival rate, disease-free sur-rival rate and local control rate were 93.1%, 88.1% and 94.6% at 2-year, and 75.7%, 62.3% and 85.6% at 5-year,respectively. The independent prognostic factors were lymph node metastasis and positive surgical margin for overall survival, positive surgical margin for local control, and stage, uterine body invasion and positive surgical margin for disease-free survival. Sixteen patients ( 14% ) had distant metastasis, and the most common sites were the lung,inguinal region,bone,liver and brain. According to RTOG grading sys-tem, the incidence of late gastrointestinal side effects of grade 1,2 and 3 was 11.4%, 11.4% and 3.5%. The corresponding genitourinary side effects were 14.0% ,6.1% and 0.9%, respectively. The incidence of leg lymphedema was 7 % . Conclusions Post - operative radiotherapy can achieve good local control in cervical cancer with acceptable late side effects. Distant metastasis is the main cause of death.

OBJECTIVETo study the effect of growth hormone (GH) combined with Radix Dipsaci on the body growth and the bone mineral content (BMC) of hypophysectomized rats.

METHODSThe GH deficiency rats model was established using the hypophysectomized operation through the skull and the throat. Qualified rats were divided into the sham-operation group (n = 15), the negative control group (n = 13), the GH intervention group (n = 13), and the GH combined with Radix Dipsaci group(n = 12). GH (0.25 mg/kg) was subcutaneously injected from the cervical part in the GH intervention group and the GH combined with Radix Dipsaci group at the same time, while equal volume of normal saline was injected to the rest groups. 0.7 mL/100 kg Radix Dipsaci was given by gastrogavage to the GH combined with Radix Dipsaci group at the same time, while equal volume of normal saline was given by gastrogave to the rest groups. The body weight, the tail length, and the body length were measured during the intervention period. Blood was withdrawn after 14-day intervention. The femoral bone and the tibial bone were taken out. The levels of GH, insulin-like growth factor 1 (IGF-1), alkaline phosphatase (ALP), and osteocalcin (OC) were measured. The width of the tibial epiphyseal plate was measured. The bilateral femur bone mineral density (BMD) and BMC were measured using the dual energy X-ray absorptiometry.

RESULTSThe body weight, the body length, the length of the femoral bone, the length of the tibial bone, the width of the epiphyseal plate, the levels of the GH, IGF-1, ALP, and OC increased in the GH intervention group and the GH combined with Radix Dipsaci group after 2-week intervention, showing statistical difference when compared with the model group (P < 0.01). But there was no statistical difference in the tail length though it also increased (P > 0.05). There was insignificant difference in the aforesaid indices between the two groups (P > 0.05). Compared with the model group, the BMD of the GH combined with Radix Dipsaci group increased with statistical difference (P < 0.01). Compared with the model group, the BMC of the GH intervention group and the GH combined with Radix Dipsaci group increased with statistical difference (P < 0.01). It was highest in the GH combined with Radix Dipsaci group (P < 0.01).

CONCLUSIONSGH combined with Radix Dipsaci showed unobvious effect on promoting the growth. But it could elevate BMD and BMC, and improve the bone metabolism.

Animals ; Bone Development ; drug effects ; Bone and Bones ; metabolism ; Dipsacaceae ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Growth Hormone ; pharmacology ; Hypophysectomy ; Male ; Rats ; Rats, Sprague-Dawley

Objectives To study the level and development of serum specific antibody against severe acute respiratory syndrome coronavirus(SARS-CoV)of different populations in SARS pestilence district after SARS outbreak in a general hospital.Discuss SARS sub-clinical infection and protective action of the IgG antibody.Methods Seroepidemiology method,enzyme-linked immunosorbent assay (ELISA)and indirect immunfluorescence assay(IFA)were employed to investigate the changing level of serum antibody to SARS-associated coronavirus in non-SARS population in SARS pestilence district during and after SARS outbreak.The development of IgM and IgG antibody in patients with SARS in 6 weeks after the onset of SARS was studied qualitatively.The level changing of IgG antibody in con- valescent patients with SARS in 82 weeks after the onset was observed dynamically.Results The ELISA test outcome of IgG antibody was negative in 200 non-SARS people who were random samples of normal mass in SARS pestilence district and common community.The positive rate was 0.41% in 487 SARS high risk population tested by ELISA,but showed negative when retested by IFA.The A value level of IgG antibody existed significant difference in non SARS mass during and after SARS outbreak and the later's was higher them the former's(P

Objective To explore effect of endogeneous gangliosides(Gls) and integrin ?2?1 on protein phosphotyrosine expression of pp125 focal adhesion kinase (pp125FAK) after adhesion of SK-N-SH neuroblastoma cells to collagen(Col).Methods SK-N-SH cell line with high expression of integrin ?2?1 was cultured in presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol(D-PDMP).Effect of endogeneous Gls,anti-?2 and anti-?1 monoclonal antibody on protein phosphotyrosine expression of pp125FAK during adhesion of SK-N-SH cells to Col were determined by immunoprecipitate and Western blotting.Results After 6 days,endogenous Gls in cells were almost depleted.Gls-depletion,anti?2 and anti-?1 monoclonal antibody were able to decrease pp125FAK expression of SK-N-SH cells adherent to Col respectively.GD2,the major component of neuroblastoma cell Gls could reco-ver pp125FAK expression to a certain degree.Conclusions Endogenous tumor Gls regulate protein phosphotyrosine expression of pp125FAK during adhesion of neuroblastoma cells to Col.It is suggested that tumor Gls may increase signal transduction of tumor cell integrin ?2?1 by increasing tyrosine phosphorylation of pp125FAK.

Objective To investigate the expression and variation of MIP 1β,MIP-2,and IL-12p70 in mice with bloodstream infection caused by 4 kinds of bacteria.Methods CD-1 (ICR) mouse models of bloodstream infection with Staphylococcus aureus (S.aureus),Enterococcus f aecalis (E.f aecalis),Escherichia coli (E.coli),and K lebsiella pneumoniae (K.pneumoniae) were established.After mice in each trial group and PBS control group were infected by bacteria for 0.5h,1h,3h,6h,12h,24h,and 48h,concentrations of MIP-1β,MIP-2,and IL-12p70 were detected by Luminex liquid suspension chip system.Results Concentrations of MIP-1β increased significantly 1h after bacteria was in blood,S.aureus,E.faecalis,E.coli,K.pneumoniae,and control groups were (134.5 ± 18.3),(61.5 ± 15.4),(3 354.0 ±809.0),(6 888.4 ± 1 100.2),and (28.9 ± 4.6) pg/mL respectively;the peak values of IL-12p70 were (389.3 ± 118.1),(127.6 ± 10.0),(42.2 ± 3.5),(62.8 ± 8.4),and (4.8 ± 0.3) pg/mL respectively.Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were significantly higher than other trial groups and control group (all P＜0.01),while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than E.coli,K.pneumoniae,and control groups (all P＜0.01).Conclusion Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were both significantly higher than those in S.aureus and E.faecalis groups,while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than those in E.coli and K.pneumoniae groups.The combination detection of multiple cytokines or chemokines are valuable in predicting gram-positive or gram-negative bacterial infection,and can provide basis for treatment of early infection.

Adenocarcinoma ; genetics ; Adult ; Age Factors ; Aged ; Base Sequence ; Carcinoma, Squamous Cell ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Sequence Deletion ; Smoking ; genetics