1.Noncompetitive antagonists and agonists of glutamate receptor regulate filopodia formation in epidermal cells and melanosome transfer: an experimental study
Chinese Journal of Dermatology 2013;46(12):871-876
Objective To investigate the regulatory effect of glutamate signaling pathway on filopodia formation in epidermal cells and on melanosome transfer.Methods Epidermal melanocytes and keratinocytes were isolated from human foreskin and subjected to subculture.After two to three passages of subculture,the melanocytes and keratinocytes were cultured alone or in combination with or without the presence of MK801 (an antagonist of N-methyl-D-aspartic acid (NMDA) receptor) of 100 μmol/L,or NMDA (the activator of NMDA receptor) of 100 μmol/L,for 24 hours.The melanocytes irradiated with UVB at 311 nm served as the control.Scanning electron microscopy was used to observe the appearance of filopodia and dendrites of melanocytes and keratinocytes.Melanosome transfer was visualized under confocal laser scanning microscopy after double immunofluorescent staining.Results Although no obvious changes were observed in the number of dendrites in monocultured melanocytes after treatment with MK801 or NMDA for 24 hours,dendrites became thinner at the terminus and longer with a decrease in the number and length of filopodia after MK801 treatment,but thicker and shorter with an increase in the number and length of filopodia after NMDA treatment compared with untreated monocultured melanocytes.In the coculture system,filopodia were observed between the untreated melanocytes and keratinocytes,and the number of filopodia in melanocytes was larger in the side adjacent to keratinocytes than in the opposite side.Compared with the untreated coculture system,the number of both filopodia connecting melanocytes and keratinocytes and filopodia extending from melanocytes to keratinocytes decreased in the coculture system after treatment with MK801 of 100 μmol/L,but increased after treatment with NMDA of 100 μmol/L,for 24 hours.Melanosomes were found in keratinocytes cocultured with melanocytes without treatment,which were decreased in number after 24-hour treatment with MK801 of 100 μmol/L,but increased in number and even present in keratinocytes nonadiacent to melanocytes after 24-hour treatment with NMDA of 100 μmol/L.Conclusion Glutamate signaling pathway may modulate the transfer of melanosomes from melanocytes to keratinocytes via modulating the appearance of melanocyte dendrites and formation of filopodia.
2.Clinical and Experimental Observation on Treatment of Psoriasis with Radix Astra gali Injectio
Xiaoming LIU ; Xin QI ; Zhiqi SONG
Chinese Journal of Dermatology 1994;0(02):-
Objective T o observe the therapeutic effect of Radix Astragali Injectio on psoriasis and it s possible pharmaceutical mechanism. Methods Two groups of patients with psoriasis vulgaris were treated with either comprehensive regi men or comprehensive regimen plus Radix Astragali Injectio. The clinical respons es in both groups were compared. Meanwhile, the effects of Radix Astragali Decoc tion and Injectio on proliferation of vaginal epithelium, PCNA expression, diffe rentiation of tail scale epidermis and plasma ET-1 were assessed in mouse mode l. Results It was shown that psoriatic lesions began to fade significantly earli er in Radix Astragali group than in routine comprehensive treatment group. The c linical cure rate was significantly higher in Radix Astragali group also. Animal experiment indicated that Radix Astragali Injectio had effects on all 4 indices mentioned above and the effects of the Injectio was stronger than that of the D ecoction. Conclusions Radix Astragali Injectio is effective for psoriasis vulgar is. Its therapeutic effects may be explained by blocking multiple pathogenetic l inkage.
3.Histological observation on the urinary organs in normal Meriones Unguieulataus
Wenjie ZHAO ; Zhiqi SONG ; Guangyu CHENG ; Deming ZHAO ; Lifeng YANG
Chinese Journal of Comparative Medicine 2014;(12):24-26
Objective Mongolian gerbil can make themself urine concentration for saving water and adapt to the harsh desert environment, due to their very unique moisture control system in the body.Methods Mongolian gerbil is resistant to drought on account of their special kidney. Histology of the kidney, ureter and bladder in Meriones Unguiculataus were observed by light microscopy using HE staining.Results The results showed that compared with rats and mice, the Mongolian gerbils have more developed distal tubules, and well developed inner renal medulla.Conclusions We hope that the findings of this study enrich our understanding of the histology of urinary system in Mongolian gerbils and provide support for the laboratory animalization of this animal.
4.Morphea complicated by Hashimoto's thyroiditis in two sisters
Jinpeng LIU ; Xiaohong YU ; Guoling YANG ; Zhiqi SONG
Chinese Journal of Dermatology 2015;48(4):248-251
Morphea complicated by Hashimoto's thyroiditis is reported in two sisters.Case 1:a 64-year-old female presented with skin rashes on the anterior neck,trunk and bilateral anterior shins for 5 years,itching skin rashes on the perineum for 4 years,and Hashimoto's thyroiditis for 9 years.Physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Dermatological examination showed pink patches on the neck and breast,sclerosis and atrophy of skin over the back,porcelain-white patches on the perineum.Histopathological findings suggested the diagnosis of morphea on the breast and lichen sclerosus et atrophicus on the perineum.Case 2:a 55-year-old female,who was the younger sister of case 1,suffered from gradual sclerosis and atrophy of skin in the left inframammary region and abdominal region for 4 years,as well as Hashimoto's thyroiditis for 3 years.Similarly,physical examination revealed grade 1 enlargement of firm thyroid gland without exophthalmos or pretibial myxedema.Hypopigmentation,sclerosis and atrophy of skin were observed in the left inframammary region,abdominal region and central back region.Histopathological examination suggested a diagnosis of morphea.According to the clinical and histopathological manifestations,periodic acid-Schiff staining and thyroid gland function test results,the 2 cases were both diagnosed as morphea complicated by Hashimoto's thyroiditis.
5.Effects of curcumin on the AKT/mTOR signaling pathway in human melanoma cell lines A375 and C8161
Xiaodong HAN ; Youyou ZHOU ; Siwen ZHENG ; Zhen LI ; Zhiqi SONG
Chinese Journal of Dermatology 2015;48(6):382-386
Objective To explore molecular mechanisms underlying the in vitro counteracting effect of curcumin on malignant melanoma.Methods Cultured A375 and C8161 human melanoma cells were cultivated in vitro,and randomly divided into several test groups and a control group to be treated with different concentrations of curcumin and dimethyl sulfoxide respectively for different durations.Then,methyl thiazolyl tetrazolium (MTT) assay,Transwell assay,flow cytometry and Western blot were performed to evaluate the effect of curcumin on the proliferation,invasion and cell cycle of,as well as expressions of AKT/mTOR signaling pathway-related proteins in A375 and C8161 cells respectively.Statistical analysis was carried out by using t test.Results MTT assay showed that the treatment with curcumin of 5-35 mg/L for 24-96 hours significantly inhibited the proliferation of both A375 and C8161 cells compared with that with dimethyl sulfoxide (all P < 0.001),and the inhibitory effect was in a dose-dependent manner within the range of 5-15 mg/L for A375 cells and within the range of 5-10 mg/L for C8161 cells,and in a time-dependent manner from 0 to 48 hours for both cells.After treatment for 24 hours,the 50% inhibitory concentration (IC50) of curcumin against A375 cells and C 8161 cells was 10 mg/L and 5 mg/L respectively.Transwell assay demonstrated that the invasion of A375 and C8161 cells was significantly suppressed by 72-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively (both P < 0.001).Flow cytometry showed that the cell cycle of A375 and C8161 cells was arrested at G2/M phase after 24-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively,with significant differences in the proportion of A375 cells and C8161 cells in G2/M phase between the test group and control group (A375 cells:35.00% ± 3.54% vs.120.80% ± 7.46%,P< 0.001;C8161 cells:19.33% ± 4.04% vs.85.00% ± 9.53%,P < 0.001).Western blot revealed that the expressions of AKT/mTOR signaling pathway-related proteins were decreased in A375 and C8161 cells after 24-hour treatment with 10 mg/L and 5 mg/L curcumin respectively.Conclusion Curcumin can inhibit the proliferation and invasion of A375 and C8161 cells,likely by blocking cell cycle and inhibiting activation of the AKT/mTOR signaling pathway.
6.Design of 16-channel EEG signals preprocessor
Xijing JING ; Jianqi WANG ; Fei JI ; Zhiqi SONG
Chinese Medical Equipment Journal 1989;0(04):-
In order to check out the cortical EEG signals of conscious and/or anesthetized rats, a 16-channel analog signals preprocessor is developed. This preprocessor can amplify and filter the primal EEG signals to transform them into ones with proper altitudes and high SNRs, and then they will be sent to the A.D converter. Some measures are taken in view of the signals' low frequencies, low SNRs and background with strong interference. When high gain made, such performances of the circuit are emphasized as low noise, low offset voltage, low gain error and heightened CMRR.
7.Glutamate receptor signaling pathway in melanoma cells and its effect on cytoskeleton protein
Ping LU ; Yusen ZHOU ; Wanrong CHEN ; Lujuan SHAN ; Junfeng LI ; Zhiqi SONG
Chinese Journal of Dermatology 2009;42(8):575-578
Objective To investigate the effects of glutamate receptor signaling on melanoma cell dendrite morphology and cytoskeleton protein. Methods A metastatic human malignant melanoma cell line WM451LU was cultured and transfected by recombinant adenovirus vector carrying a cDNA encoding microtubule-associated protein 2a (MAP2a). MK-801, an antagonist of N-methyl-D-aspartate receptor (NMDAR), and CPCCOEt, an antagonist of metabotropie glutamate receptor 1 (mGluR1), were used to treat transfected or untrausfeeted WM451LU cells. Confocal microscopy and three dimensional atomic force microscopy were used to assess subcellular location of NMDAR2A, mGluR1 and MAP2a as well as the dis-tribution of α-tubulin in and dendrite morphology of WM451LU cells. The proliferation of WM451LU cells was estimated by cell survival growth curve. Results Confocal laser microscopy revealed that NMDAR2A, mGluRl and MAP2a were mainly co-localized in melanoma cell dendrites. Both MK-801 and CPCCOEt increased the density of microtubules in cell dendrites and dendritic branching of WM451LU cells, and both effects of MK-801 and CPCCOEt were enhanced by the expression of MAP2a. Furthermore, the proliferation of WM451LU cells was significantly inhibited by MK-801 of 100 μmol/L and CPCCOEt of 10 μmol/L. Conclusions In melanoma cells, glutamate receptors may participate in the development of dendrites, and anta- gonists of glutamate receptors could inhibit the proliferation of melanoma cells.
8.Method for the improving of point-of-care HBA1c results harmonization
Rui ZHOU ; Zhiqi GAO ; Qing TONG ; Chang ZUO ; Zhixin SONG ; Qingtao WANG
Chinese Journal of Laboratory Medicine 2015;(5):310-312
Objective To establish a new traceability pathway of point-of-care testing ( POCT ) of HBA1c by using commutable quality controls, in order to improve the accuracy of POCT HBA1c and the harmonization of testing results with those of central laboratories.Methods The study was about measurement traceability. Human frozen whole blood samples with IFCC assigned values were used to calibrate the G8 HBA1c Variant in June, 2013.According to the CLSI EP9-A2-IR guideline, 50 patient samples and 2-level commercial QC samples were then analyzed by G8 system and DCA Ventage system.The best fitting curves for fresh patient samples and the commercial QC materials were established separately. The patient results tested on the DCA Ventage were modified and verified.Paired t-test and Passing Bablok linear regression were used.Results The linear equation of DCA/G8 before calibration was Y=0.899 5X+0.389 1(R2 =0.991 0).Calibration by fresh patient samples reduced the mean bias of DCA/G8 from -0.40%±0.34%to 0.00%±0.29%.Calibration by QCs reduced the mean bias to 0.15%±0.29%.The linear correlation established by quality controls was stable, which made the bias was lower between DCA and G8 in the consequent six runs.Conclusions The accuracy and the traceability of POC testing could be realized by using commutable QC materials traceable to IFCC assigned values.Through this method, POC testing can become more comparable to the results of clinical laboratory HBA1c instruments.
9.Results analysis of blood infectious indicators in 331 968 outpatients and inpatients
Zhiqi WU ; Huiqing HUANG ; Fang NI ; Weijuan SONG ; Erfu XIE ; Huaguo XU
International Journal of Laboratory Medicine 2015;(15):2133-2134
Objective To analyze the results of detection of infectious indicators of patients from the first affiliated hospital of Nanjing Medical University in recent 5 years ,and to provide a scientific basis for the control and prevention of infectious diseases . Methods The patients with clinical data from January 2010 to April 2014 were retrospectively analyzed ,then the infectious indica‐tors of all the subjects were detected and analyzed .Results HIV positive rate was between 0 .043% to 0 .061% ,positive rate of HCV was between 1 .07 to 1 .41% ,positive rate of TP was between 2 .01% to 2 .17% .HBsAg positive rate in 2010 was 8 .36% ,the positive rate was 7 .81% in 2014 .HBsAb positive rate in 2010 was 35 .36% ,positive rate was 50 .96% in 2014 .Conclusion Effec‐tively cut off the route of transmission could prevent the further spread of infectious disease .
10.Glutamate-mediated signaling pathway regulates the invasion and growth of malignant melanoma
Lili LI ; Lujuan SHAN ; Yuan ZHANG ; Chuanzhou GAO ; Haiqin GAO ; Wenting GAO ; Yuejian LIU ; Zhiqi SONG
Chinese Journal of Dermatology 2011;44(3):186-190
Objective To investigate the action mechanism of glutamate-mediated signaling pathway in malignant melanoma. Methods WM451LU melanoma cells in log phase were classified into 6 groups, negative control group treated with PBS (100 μl), MK801 group treated with the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (100 μmol/L), CPCCOEt group treated with non-competitive metabotropic glutamate receptor 1 (mGluR1) antagonist CPCCOEt, MAP2 group transfected with adenovirus vector containing microtubule associated protein 2a (Ad-MAP2a), MK801 + MAP2 group treated with MK801 of 100 μmol/L and transfected with Ad-MAP2a, CPCCOEt + MAP2 group treated with CPCCOEt of 10 μmol/L and transfected with Ad-MAP2a. Western blot was performed to detect the expression of an ionotropic glutamate receptor, i.e., N-methyl-D-aspartate receptor type 2A (NMDAR2A) in WM451LU cells transfected with Ad-MAP2a. Scratch motility assay and cell invasion assay were conducted in vitro to detect the changes in migration and invasion ability of WM451LU cells after treated with Ad-MAP2a, MK-801, CPCCOEt alone or in combination. In vivo study was carried out to compare the inhibitory effect of the above treatments on melanoma. Results Western blot revealed a decrease in the expression of NMDAR2A in WM451LU cells after transfected with Ad-MAP2a. The scratch motility assay showed that the number of migrating cells per high power field was 117.04 ± 2.76 in MAP2 group,107.64 ± 6.50 in MK801 group,97.36 ± 4.79 in CPCCOEt group, 43.28 ± 3.02 in MK801 + MAP2 group,30.76 ± 3.97 in CPCCOEt + MAP2 group,significantly different from that in the negative control group (152.3 ± 5.75,all P < 0.01 ). Cell invasion assay demonstrated that the average number of invading cells per high power field in the negative control was significantly higher than that in MAP2 group, MK801 group, CPCCOEt group, MK801+MAP2 group and CPCCOEt + MAP2 group (170.43 ±8.72 vs. 98.26 ± 3.84, 97.22 ± 5.54, 112.23 ± 7.21, 42.89 ± 5.06, 58.25 ± 6.68, P < 0.05, 0.05, 0.05, 0.01and 0.01, respectively).A significant decrease was observed in the average volume of experimental melanoma in mice of MAP2 group, MK801 group, MK801 + MAP2 group, CPCCOEt group and CPCCOEt + MAP2 group compared with the negative control group (224.02 ± 46.19 mm3, 160.33 ± 33.91 mm3, 91.49 ± 21.48 mm3,202.30 ± 52.37 mm3, 111.13 ± 69.81 mm3 vs. 342.70 ± 60.92 mm3, all P < 0.01 ). Conclusions To block the glutamate signaling pathway in vitro can inhibit the invasion and migration of melanoma cells, and to block the pathway in vivo can inhibit the growth of malignant melanoma and alter the morphology of melanoma cells.