Objective To establish a HPLC method for the content determination of Chlorogenic acid and Hyperoside in the leaves of acanthopanax senticosus from different areas.Methods The samples were separated on Hedera? ODS-2 C18 column,using acetonitrile-0.4% phosphoric acid(17∶83) as mobile phase,the detection wavelength was 360 nm.Result Chlorogenic acid and Hyperoside showed good linear relationship over the ranges of 10.2～306 ?g/mL(r =0.999 5) and 10.4～312 ?g/mL(r =0.999 8),respectively.The average recoveries were 97.14% and 98.21% with RSD at 1.65% and 1.09%,respectively.The contents of Chlorogenic acid and Hyperoside were different obviously in samples from different areas.Conclusion This method is simple,rapid and sensitive,and it can be used for the further development and the quality control of the leaves of acanthopanax senticosus.

Objective To investigate the antimicrobial properties of human amniotic homogenate supernatant (HAHS) in order to found a theoretical base for the use of human amniotic membrane in ophthalmological field. MethodsFresh human amniotic membranes were used to make HAHS and acellular amniotic membranes. Then, we observed their antimicrobial effects and antimicrobial spectrums, compared the antimicrobial capacity with 10 commonly used antibiotics in eyes, and investigated the effects of time, temperature and pH value on the antimicrobial capacity. Finally, transmission electron microscopy (TEM) was used to explore the possible targeting site of the antibiosis. ResultsHuman amnion membrane contained antimicrobial components locating in its epithelial cells. HAHS had a broad antimicrobial spectrum and was steady in nature. Its antimicrobial capacity was stronger than those of sulfasulfonamide, chloromycetin and cefuroxime sodium. TEM indicated that the antimicrobial effect were exerted through plasma membrane of microorganism. ConclusionHAHS can be an effective and convenient treatment for ocular surface infectious diseases. Traditional amnion transplantation should employ fresh human amniotic membranes containing complete epithelial lamina to reconstruct the ocular surface.

Objective To investigate the changes in histological characteristics and collagen content in uterosacral and cardinal ligaments of pefimenopausal women in relation to relaxation of pelvic supports.Methods Twenty-eight subjects undergoing hysterectomies were selected,in which 14 cases were perimenopausal women with relaxation of pelvic support as the relaxation group and 14 women at perimenopansal age with leomyoma,cervical cancer,adenomyosis as the control group.Samples of cardinal ligaments and uterosacral ligaments were obtained at hysterectomies,and the tissues were shced and stained bv Masson's trichrome technique.Histological characteristics of the samples were studied and immunohistochemistry assay was applied to demonstrate the contents of collagen types I andⅢ.Results (1)The collagen in uterosacral ligaments and cardinal ligaments were stained blue by tlle MaSson's trichome technique.In comparison to the control group,the relaxation group had milder positive stains of the collagen and the stains were distributed in unequal intensities.Collagen content was arranged in loose pattern.Focal arrangement of the collagen was dense but fragmented.Collagen fibers were atrophic. (2)In immunohistochemistry assay and image analysis,collagen was positive in light to deep brown areas.In the relaxation group,positive units of collagen types I and III in cardinal ligaments were 13.8±2.1 and 9.6±2.4 respectively.Positive units of coHagen types I and Ⅲ of cardinal ligaments in the control group were 27.4 ±3.5 and 17.7 ±4.0 respectively.Difierences between these two groups were statistically significant (P

Objective To investigate the effects of 5-azacytidine on radiation-induced epithelialmesenchymal transition in rat alveolar type Ⅱ epithelial cell line (RLE-6TN) and explore their working mechanisms,and to provide experimental evidence for the potential drug-based treatment of radiationinduced pulmonary fibrosis.Methods RLE-6TN cells were cultured in vitro and divided into four groups according to the experimental purposes:control group (C),radiation group (R),5-azacytidine group (A),and radiation followed by 5-azacytidine group (R + A).The microstructural changes in cells were determined by transmission electron microscopy.Inverted phase-contrast microscopy revealed the morphological changes in cells.The mRNA expression levels of E-cadherin and α-SMA were measured by quantitative real-time polymerase chain reaction (qRT-PCR).The protein expression levels of E-cadherin,GSK3 β,and p-GSK3 β (Ser9) were measured by Western blot.The one-way analysis of variance was used for pairwise comparison.Results The cells in group R became spindle-like.Similar morphological changes were not observed in cells in group R + A.Osmiophilic lamellar bodies disappeared at last in cells in group R.RT-PCR results showed that compared with group C,group R had a significantly lower mRNA expression level of E-cadherin ((0.23 ± 0.06) vs.(1.00 ± 0.00),P =0.002)) and a significantly higher mRNA expression level of α-SMA ((2.91 ± 0.01) vs.(1.00 ± 0.00),P =0.000)).However,compared with group R,group R + A had a significantly higher mRNA expression level of E-cadherin ((0.47 ± 0.05) vs.(1.00 ± 0.00),P =0.024)) but a significantly lower mRNA expression level of α-SMA ((2.50 ± 0.02) vs.(1.00 ±0.00),P =0.037)).The results of Western blot showed that the protein expression level of Ecadherin was significantly reduced ((0.07 ± 0.01) vs.(0.48 ± 0.02),P =0.028)),while the protein expression level of p-GSK3β was significantly increased in Group R than in Group C ((0.85 ± 0.04) vs.(0.23 ± 0.03),P =0.031)).However,compared with group R,group R + A had a significantly lower protein expression level of E-cadherin ((0.25 ± 0.00) vs.(0.07 ± 0.01),P =0.024)) and significantly less up-regulation of the protein expression level of p-GSK3β ((0.39 ± 0.03) vs.(0.85 ± 0.04),P =0.014)).Conclusions X-ray radiation can induce the epithelial-mesenchymal transition in epithelial cells.5-azacytidine suppresses radiation-induced epithelial-mesenchymal transition by inhibition of the activity of p-GSK3β in RLE-6TN cells.

To investigate the effect of the anti-endotoxic part of Radix Isatidis on the expression of moesin mRNA in murine tissues induced by lipopolysaccharide (LPS), the sample solution of F(022) part from Radix Isatidis was intraperitoneally administered to experimental mice, and the lipopoly-saccharide (LPS) were injected into the tail vein, and then the tissues of liver, kidney and spleen were colleted and cut into slices. The mRNA was detected by moesin mRNA hybridization in situ. The staining results were observed under microscope. It was found that moesin mRNA expression was increased in the tissues of liver, kidndy and spleen in mice treated with LPS, while in the mice pre-treated with F(022) part from Radix Isatidis, the LPS-induced moesin mRNA expressions in these tissues were inhibited in a dose-dependant manner. Our study showed that F(022) part from Radix Isatidis can inhibit the LPS-induced expression of moesin mRNA in the tissues of liver, kidney and spleen in mice.

Objective To investigate the changes of IL-8 in peripheral blood of patients with pustular psoriasis after acitretin treatment.Methods Dual antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to measure the IL-8 levels in the peripheral blood of 30 patients with pustular psoriasis and 30 patients with psoriasis vulgaris before and after treatment,and in 30 normal human controls.The relationship between the IL-8 level and disease severity was assessed for patients with pustular psoriasis.Results After acitretin treatment,the condition was improved greatly in patients with pustular psoriasis,together with a significant decrease in the level of IL-8 in the peripheral blood(57.07 ± 12.02 pg/ml vs.96.84 ± 14.68 pg/ml,P ＜ 0.05).Conclusion IL-8 may be involved in the therapeutic mechanism of acitretin in pustular psoriasis.

Objective To investigate the expression of granulocyte macrophage-colony stimulating factor(GM-CSF)in skin lesions and sera of patients with pustular psoriasis and psoriasis vulgaris.Methods Tissue specimens were obtained from the lesions of 15 patients with pustular psoriasis,15 patients with psoriasis vulgaris and 15 normal human controls.Immunohistochemistry and dual antibody sandwich enzyme-linked immunosorbent assay(ELISA)were carried out to detect the levels of GM-CSF in the tissue and serum specimens from the patients and normal human controls,respectively.Results Significantly higher levels of GM-CSF were observed in the tissue and serum specimens from patients with pustular psoriasis and psoriasis vulgaris compared with the normal controls(all P ＜ 0.01),as well as in those from the patients with pustular psoriasis compared with the patients with psoriasis vulgaris(both P ＜ 0.01).Conclusion GM-CSF may be involved in the pathogenesis of psoriasis.

Objective:To explore the involvement of endoplasmic reticulum molecular chaperone GRP78 in the impairment of inner ear consistented with the mimetic aging model.Method:Twenty-four Wistar rats were randomly divided into two groups. model group was in duced by daily hypodermic injection of 10% D-galactose (800 mg·kg~(-1)·d~(-1)) for 8 weeks and the control group was given saline accordingly. Spatial learning and memory was measured by Morris-Water-Maze. Colorimetry was used to analyze superoxide dismutase (SOD) and malondialdehyde (MDA) extracted from inner ear tissue. Hearing threshold of rats were detected with Auditory brainstem response (ABR).In addition, expression of GRP78 in the inner ear was detected by immunohistochemistry,RT-PCR and Western blot. The control group was studied parallel.Result:The escape latency in the model group injected with D-galactose was markedly longer than that in the control group.accordingly ,the changes of SOD and MDA were more significant in the model group, the difference between two groups was significant(t-test,P<0.01). the variation of ABR in two groups was observed, There was no statistically difference of the hearing in the model group compared with the control group(P>0.05). The expression of GRP78 was significantly different between two groups ,which is increased in the inner ear tissue of model group(P<0.01).Conclusion:The impairment of inner ear tissue partly dued to the oxidative stress in the model, which was induced by D-galactose.and endoplasmic reticulum molecular chaperone was thought to contribute to the impairment mechanism of inner ear in mimetic aging model.

Objective To identify and quantitate the conjunctival aerobic bacterial flora from healthy people of 3 age groups in Chongqing city.Methods Samples taken with moist swabs from the lower fornix of both eyes of 33 children(9.85?0.57 years old),31 youths(21.23?0.88 years old)and 30 elders(72.97?4.94 years old)were given bacterial cultures.Positive ones were identified and colony forming units(CFUs)were counted.Bacterial quantity was expressed as the number of CFUs divided by the weight of each sample measured before and after collection.Results The elders showed a higher isolation frequency(93.33%)than children(45.45%)and youths(50.00%)which had statistically difference(P

Objective To investigate the value of external counterpulsation and traction therapy on patients with vertebral artery insufficiency syndrome by use of transcranial Doppler (TCD). Methods Based on the criteria of clinical diagnosis, 86 patients with vertebral artery insufficiency syndrome and 60 healthy persons were examined by EME 4004 type TCD. The peak velocity (Vp) and mean velocity (Vm) of blood flow of the vertebral artery (VA) and basilar artery (BA) were detected and analyzed before and after rehabilitation treatment. Results The blood flow velocities of the vertebrobasilar artery (VBA) of the patient group were much lower than that of the control group ( P 0.05). Conclusion TCD examination while rotating neck could provide valuable information for the diagnosis and rehabilitation assessment of patients with vertebral artery insufficiency syndrome. External counterpulsation and traction therapy are effective in the treatment of the vertebral artery insufficiency syndrome.