AIM: Test the character of Electrooculogram (EOG) in normal subjects so as to obtain reference values.METHODS: By using Vision Monitor visual evoked response imaging system, the EOG was recorded on 60 normal subjects (73 eyes).RESULTS: EOG under the condition of normal pupil was recorded in normal subjects according to ISCVE standard. The dark trough potential was (701.8±265.1)μV, the light peak potential was (1255.0±447.7)μV, the Arden ratio (light peak /dark trough ratio)was 180%±21%.CONCLUSION: Our study reflected the spatial characteristics of electrooculogram in normal subjects,provided reliable normal reference values for clinical research.

Breast cancer is called "Ruyan" in literature of traditional Chinese medicine. We synthesized the ancient and contemporary discussions and raised the theory that "Duxie" (poisonous pathogenic factor) is the etiological factor and pathologic product through the whole course of breast cancer. "Liuyin Fudu" (latent poison of six exogenous pathogenic factors) and "Qiqing Yudu" (stagnant poison of seven emotions) are the main etiological factors affecting the breast cancer occurrence. "Aidu Neisheng" (internal product of cancer poison) is the essential change in breast cancer occurrence. "Tandu Yujie" (stagnation of phlegm, poison and blood stasis) is the essential pathogenesis of the breast cancer's development. "Yudu Weiqing" (vestigial poison) is the main pathogenesis of breast cancer after operation. "Yudu Pangcuan" (vestigial poison invasion elsewhere) is the key pathogenesis of recurrence and metastasis after operation. "Sanjie Jiedu" (dispersing accumulation and detoxification) is an important therapeutic principle in breast cancer's treatment after operation. The "Tandu Yujie" pathogenesis theory and "Sanjie Jiedu" therapeutic principle developed the theory about breast cancer in traditional Chinese medicine, and have some clinical application value.

OBJECTIVE: To observe the effect of Ruyiping, a traditional Chinese compound herbal medicine composed of 5 Chinese herbs for removing toxic materials and dissipating nodules from Runing II, another traditional Chinese compound herbal medicine for treating breast cancer, in preventing recidivation and metastasis in breast cancer patients after operation. METHODS: Eighty patients with breast cancer after operation were randomly divided into Ruyiping group and Runing II group, and prescribed Ruyiping and Runing II on the basis of chemotherapy, radiotherapy and endocrine therapy respectively for two years. RESULTS: There were two patients with metastasis and three patients lost to follow-up in Ruyiping group and three and two in Runing II group. The recidivation and metastasis rates were 5.41% and 7.89% respectively. The difference between the two groups was not statistically significant (P>0.05). The difference of disease-free survival time between the two groups was also not statistically significant. CONCLUSIONS: The effect of Ruyiping in preventing recidivation and metastasis is similar to that of Runing II. Ruyiping is the essential component of Runing II for preventing recidivation and metastasis. The result provides some clinical evidences for the theory that "Yudu Pangcuan" (vestigial poison invasion elsewhere) is the essential pathogenesis of breast cancer's recidivation and metastasis and the utilization of "Sanjie Jiedu" (dispersing accumulation and detoxification) is the therapeutic principle in preventing recidivation and metastasis after operation.

A new phenethanol, (2'R)-4-(2', 3'-dihydroxy-3'-methyl-butanoxy)-phenethanol (1), along with other eleven known benzene derivatives (2-12) were isolated from the roots, stems and leaves of Clausena excavata (Rutaceae). Compounds 3 and 4 are new natural products, and compounds 5-8, 10-12 were isolated from C. excavata for the first time. Their structures were elucidated on the basis of MS, 1D and 2D NMR spectroscopic analyses including HSQC, COSY and HMBC experiments. 1 was tested for its cytotoxicities against A549, HeLa and BGC-823 cancer cell lines, and antimicrobial activities against Candida albicans and Staphylococcus aureus. The results showed that 1 did not exhibit cytotoxic and antimicrobial activities.
Benzene Derivatives ; chemistry ; Candida albicans ; drug effects ; Cell Line, Tumor ; Clausena ; chemistry ; HeLa Cells ; Humans ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Staphylococcus aureus ; drug effects

To investigate monoterpenes and sesquiterpenes of the stems and leaves of Clausena excavata, an AcOEt fraction of the methanol extract was subjected on column chromatographies including silica gel and RP-18, as well as preparative HPLC. The structures of compounds isolated were identified on the basis of spectroscopic data as excamonoterpene (1), (6R, 9S)-9, 10-dihydroxy-4-megastigmen-3-one (2), (3R, 6R, 7E) -3-hydroxy-4, 7-megastigmadien-9-one (3), (3S) -3-hydroxy-7, 8-dihydro-beta-ionone (4), (3S, 5R, 6S) -3-hydroxy-5,6-epoxy-beta-ionone (5), (6R, 9R) -9-hydroxy-4-megastigmen-3-one (6), (3S, SR) -dihydroxy-6, 7-megstigmadien-9-one(7), (-)-loliolide(8), caryolane-1, 9alpha-diol(9) and 2, 6-dihydroxyhumula-3 (12), 7 (13), 9(E)-triene (10), were isolated from the stems and leaves of C. excavata. Compound 1 is a new monoterpene, named as excamonoterpene. Compounds 2-10 were isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; methods ; Clausena ; chemistry ; Magnetic Resonance Spectroscopy ; Methanol ; chemistry ; Molecular Structure ; Monoterpenes ; analysis ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Sesquiterpenes ; analysis ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.

Culture environment is the key factor in the construction of dermal skin.It was investigated that the effects of the culture methods,including the static culture and spinner flask culture,and stir speeds on the cells proliferation,metabolism and distribution within collagenchitosan sponges.A higher cell density and specific growth rate was obtained with spinner flask culture versus static culture,especially,the 80 r/min spinner flask culture.The cell distribution in dermal substitutes from stirred culture system was more uniform than static culture,as well as that with increase of stir speeds in spinner flask.In summary,the spinner flasks culture with proper stir speed shows promise for the construction of dermal substitutes in vitro.

The effect of trypsin on the separation an subculture of the keratinocytes was investigated in this work. It was found that when 0.25% trypsin was employed for 5 minutes to separate keratinocytes, the number of active keratinocytes and the cells capable of forming colony were higher than those of other experimental conditions. The maximum attached ratio of primary keratinocytes was obtained when skin tissues were treated at 0.05% concentration of trypsin. With the increase of the trypsin concentrations, the attached ratio, attachment rate constant, and colony forming efficiency were all increased. Thus, 0.25% concentration of trypsin was recommended for separating and subculturing the keratinocytes.
Animals ; Cell Count ; methods ; Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cell Survival ; drug effects ; Keratinocytes ; cytology ; drug effects ; Mice ; Stem Cells ; drug effects ; Trypsin ; pharmacology

The effect of antioxidants on the in vitro life span of mouse keratinocytes was investigated in this work. It was found that the life span of the keratinocytes cultured in the medium supplemented with antioxidants was extended significantly. The most beneficial antioxidant used in this work was the mercaptoethanol, followed by the catalase and SOD. However, the growth rates of keratinocytes in vitro under all the experimental conditions still declined with the culture time. It was also found that the antioxidants added in the medium were also helpful to enhance the keratinocyte colony formation. In addition, the aging kinetics of the mouse epidermal keratinocytes in vitro were analyzed, and finally the aging rate constants corresponding to antioxidants used were calculated.
Animals ; Antioxidants ; pharmacology ; Catalase ; pharmacology ; Cell Division ; drug effects ; Cells, Cultured ; Cellular Senescence ; drug effects ; Keratinocytes ; cytology ; drug effects ; Mercaptoethanol ; pharmacology ; Mice ; Superoxide Dismutase ; pharmacology

The effect of the calcium ion (Ca2+) on the growth and differentiation of the mouse epidermal keratinocytes cultured in serum-free medium was investigated. It was found that the optimal level of calcium ion in the medium was about 0.2 mmol/L. Under such a culture condition the colony forming efficiency, attachment percentage, percentage of the cells with cornified envelops, and percentage of the senesced cells were measured to be about 10.8%, 30.8%, 5.1%, and 26.8%, respectively. However, the Ca2+ concentrations in the medium above 0.6 mmol/L resulted in significant differentiation and senescence of the keratinocytes, which was found to be harmful for keratinocyte growth and expansion in vitro.
Animals ; Calcium ; pharmacology ; Cell Adhesion ; drug effects ; Cell Differentiation ; Cell Division ; drug effects ; Cells, Cultured ; Cellular Senescence ; drug effects ; Epidermis ; cytology ; Keratinocytes ; cytology ; drug effects ; Mice