1.Voltage-gated potassium channel in a human skin squamous cell carcinoma cell line A431
Yuheng LIANG ; Xia XIONG ; Shihua ZENG
Chinese Journal of Dermatology 2011;44(6):430-433
Objective To investigate the role of voltage-gated potassium channel in the human skin squamous cell carcinoma A431 cells and human keratinocyte HaCaT cells. Methods MTT assay was performed to detect the effect of different concentrations of tetraethylammonium (TEA) on the proliferation of cultured A431 cells and HaCaT cells. Besides, enzyme-linked immunosorbent assay (ELISA) was conducted to detect the expression of HERG channel protein in A431 cells, HaCaT cells and normal human skin tissue.Results TEA inhibited the proliferation of A431 cells and HaCaT cells in a dose- and time-dependent manner.After treated with TEA (≥ 10 mmol/L) for 24 or more hours, the proliferation of A431 cells and HaCaT cells was obviously suppressed. A significant difference was observed in the average concentration of HERG channel protein between A431 cells, HaCaT cells and normal human skin tissue (49.7114 ± 3.55696 pg/ml, 35.7471 ±4.14696 pg/ml, 36.8857 ± 3.47810 pg/ml, all P < 0.05). Conclusions The block of voltage-gated potassium channel could inhibit the proliferation of A431 cells and HaCaT cells, and the expression of voltage-gated potassium channel seems to be higher in human skin squamous cell carcinoma cells.
2.Commissioning of intensity - modulated radiation therapy system with independent collimator
Ke ZHANG ; Jianrong DAI ; Li CHEN ; Yuheng LI ; Liang LI
Chinese Journal of Radiation Oncology 2011;20(2):164-167
Objective To perform an acceptance test for the IMRT system with independent collimator. Methods An ion chamber dosimeter were used to measure the startup characteristics of the accelerator and the absolute dose at isocenter and given characteristic points for three clinical cases ( a lower nasopharyngeal carcinoma, a lung cancer and a cervical cancer). The characteristic points represented the organs at risk or the target. A Mapeheck2 was used to measure dose maps of basic test fields and the treatment fields for the clinical cases. The basic test fields were as follows: 1 ). Symmetric fields in size of 2 cm ×2 cm, 5 cm ×5 cm, 10 cm× 10 cm, 20 cm ×20 cm, 2 cm × 10 cm, 10 cm ×2 cm, 5 cm ×20 cm and 20 cm ×5 cm;2). Asymmetric fields in size of 2 cm ×2 cm (x1 =4 cm, y1 = 10 cm;x2 = -2 cm, y2 = -8cm) and 5 cm ×5 cm (x1 = -2 cm, y1 = -5 cm;x2 =7 cm, y2 = 10 cm) ;3) A 20 cm ×20 cm composite field composed of five 20 cm× 4 cm narrow bar fields side by side. Gamma Index was used to compare calculated and corresponding measured dose distributions. When the criterion was 3% dose difference or 3 mm distance-to-agreement, the pass rate was required to be more than 90%. Results The accuracy of machine output was better than 2% when machine monitor units increased to 4. Among all basic test fields and all the treatment fields of three clinical cases, the maximal absolute dose error was -3.67%, and only the composite test field and two treatment fields of the lower nasopharyngeal carcinoma case had a pass rate slightly less than 90%, which were 83.6%, 88. 3% and 89. 7% ,respectively. For the three clinical cases the treatment delivery times were 15, 14, and 27 minutes, respectively. Conclusions The overall commissioning results are acceptable, and the system can be used in clinic.
3.Effect of PSMA7 on RB pathway in A549 cells
Xiang HUANG ; Yuheng ZHONG ; Jiayu TAN ; Xuewei XU ; Rui LIANG ; Shaoxia LIANG ; Jing ZHAO
Chinese Journal of Immunology 2017;33(4):516-519
Objective:To investigate the effect of upregulated and downregulated PSMA7 on the cell cycle and Cyclin D1,CDK4,P16,Rb of RB pathway in A549 cells.Methods:Transfected upregulated pcDNA3.1-PSMA7 vecter and downregulated pGPU6/Hygro-PSMA7-265 vecter into A549 cells,and then tested the effect of PSMA7 on the cell cycle of A549 cells by flow cytometry,and detected the protein level of Cyclin D1,CDK4,P16,Rb by Western blot.Results:Compared with the control group,the cell cycle of the A549 cells did not change significantly,and the expression of Cyclin D1,CDK4 decreased but P16,Rb increased when PSMA7 was upregulated.Compared with the control group,the proportion of phase G0/G1,G2/M of the A549 cells decreased and phase S increased,and the expression of Cyclin D1,CDK4 increased but P16,Rb decreased when PSMA7 was downregulated.There was statistical significance for those results.Conclusion:PSMA7 could affect the expression of Cyclin D1,CDK4,P16,Rb protein level of RB pathway in A549 and promoted the A549 cells into phase S when it′s downregulated.
4.Roles of complement receptor 3 on murine macrophages in recognition of Penicillium marneffei
Yongxuan HU ; Junmin ZHANG ; Sha LU ; Xiqing LI ; Yuheng LIANG ; Changming LU ; Liyan XI
Chinese Journal of Dermatology 2013;46(8):538-542
Objective To evaluate the role of complement receptor 3 (CR3) on murine macrophages in the recognition of Penicillium marneffei.Methods RAW264.7 murine macrophage cells were cultured in vitro,and divided into four groups to be cocultured with inactivated and live Penicillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5% CO2 for one hour.The RAW264.7 cells incubated with phosphate-buffered saline (PBS) served as the blank control group.Then,reverse transcription-PCR was conducted to detect CR3 mRNA expression,Western blot to measure CR3 protein expression,flow cytometry to determine phagocytosis rate,enzyme-linked immunosorbent assay (ELISA) to quantify cytokine levels in culture supernatant.Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia,subsequently,phagocytosis rate and supematant cytokine levels were determined.Data were processed by the SPSS 16.0 software,and one-way analysis of variance (ANOVA) was conducted for inter-group comparisons of these parameters.Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marneffei (both P > 0.05).The phagocytosis rate was 95.14%,89.56%,91.03% and 90.78% in RAW264.7 cells cocultured with inactivated conidia and yeast cells,as well as live conidia and yeast cells of Penicillium marneffei,respectively (P > 0.05).The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour coculture in the four coculture groups compared with the blank control group,but no statistical difference was noted among the four coculture groups in the supernatant levels of these cytokines (all P > 0.05).After coculture with inactivated Penicillium marneffei conidia,the siRNA-transfected RAW264.7 cells showed a statistical decrease in phagocytosis rate (10.89% vs.92.78%,P < 0.05) and supernatant levels of IL-2,IFN-γ IL-4 and IL-10 compared with untransfected RAW264.7 cells.Conclusions In early stage of innate immunity,CR3 on macrophages may be one of the pattern recognition receptors participating in the recognition and mediation of phagocytosis of Penicillium marneffei.It's possible that both Thl-and Th2-type cytokines,such as IL-2,IFN-γ,IL-4 and IL-10,are involved in the immune response of macrophages against Penicillium marneffei.
5.Fabrication of molecular probe targeting NRP-1 and magnetic resonance molecular imaging of gliomas in vitro
Yuheng SHAN ; Zhikong LIANG ; Yanchen LIN ; Lili DAI ; Yuliang WANG ; Yanhua GONG
Chinese Journal of Behavioral Medicine and Brain Science 2016;25(8):677-681
Objective To synthetize a novel MR molecular imaging probe named USPIO?PEG?tLyP?1,and to evaluate its value in detecting U87 cells by MR imaging. Methods USPIO?PEG?tLyP?1 was synthetized by conjugating USPIO?PEG with tLyP?1. Neuropilin?1 expression levels of glioma cell lines were detected by Western blot. The cytotoxicity of USPIO?PEG and USPIO?PEG?tLyP?1 were assessed by MTT colorimetric assay. The uptake efficiency of USPIO?PEG?tLyP?1 was measured by Prussian blue staining, transmission electron microscope and MR imaging in vitro. Results The novel MR molecular imaging probe was synthetized with an average diameter of 43.84 nm. U87 glioma cell line was screened as test subject for the highly expression of NRP?1(P<0.05). USPIO?PEG?tLyP?1 group showed much more intracellular blue particles than USPIO?PEG group after Prussian blue staining. After incubation,USPIO?PEG?tLyP?1 mainly existed in lysosme,endoplasmic reticulum and mitochondria. In vitro MRI showed that USPIO?PEG?tLyP?1 significantly enhanced the negative contrast effect compared with USPIO?PEG(P<0.01). Conclusion The decoration of tLyP?1 enhanced targeting ability of USPIO?PEG to glioma cells and MR molecular imaging can be a promising method for early diagnosis of gliomas.
6.Muscle atrophy of hand as an initial sign in a patient with protoplasmic astrocytoma:a case report and literature review
Xiaolei GUO ; Yanfeng ZHONG ; Yuheng SUN ; Honghong LIU ; Bo JIN ; Wei LIANG ; Xiaosong LI
Journal of Peking University(Health Sciences) 2003;0(06):-
We reported a rare case of protoplasmic astrocytoma presenting small muscle atrophy of the right hand as an initial sign.A 39-year-old male was admitted to hospital complaining of chronic muscle atrophy and subtle headache.Electromyography(EMG) showed brief small denervation and no signs of sensory-motor conduction impairment.CT and MRI revealed multiply expansive intracranial lesion in left hemisphere,which was highly suspected of cerebral echinococccus or Balo disease.The patient underwent surgical excision and pathological report was protoplasmic astrocytoma,with glial fibrillary acidic protein(GFAP,+++) of immunohistochemical method.We reviewed clinical features,radiological manifestations and pathology of protoplasmic astrocytoma with medical literature documents.
7.Surveillance and drug resistance analysis of Salmonella in Guangdong province in 2015
Dongmei HE ; Bixia KE ; Honghui ZENG ; Tong YANG ; Yuheng LIANG ; Hailing TAN ; Bosheng LI ; Meizhen LIU ; Changwen KE
Chinese Journal of Microbiology and Immunology 2017;37(8):611-617
Objective To analyze the serotype distribution and antibiotic resistance characteristics of Salmonella strains isolated in Guangdong province for better understanding the condition of Salmonella infection in patients with diarrhea.Methods Fecal samples collected from patients with diarrhea in Guangdong province were used to isolate Salmonella strains.Biochemical analysis was performed to identify these isolated strains.Serotyping and antimicrobial susceptibility testing were carried out for further analysis of the isolated Salmonella strains.Results The rate of Salmonella infection was 7.64%in 2015, and the male to female patient ratio was 1.52∶1.A total of 2 377 patients of all age groups were positive for Salmonella infection and the patients aged 0-6 years accounted for 81.74%.The isolation rate of Salmonella strains in the summer and autumn was higher than that in the winter and spring (10.73% vs 4.24%;X2=463.77, P<0.01).The Salmonella isolation rates in different areas were as follows: 16.82% in Zhuhai, 15.85% in Heyuan, 11.81% in Yangjiang, 10.68% in Jiangmen, 8.49% in Zhongshan, 8.07% in Maoming, 8.05% in Jieyang, 7.35% in Shaoguan, 6.97% in Foshan, 6.03% in Dongguan, 5.48% in Guangzhou and 0.00% in Zhanjiang.And the differences between different regions were statistically significant (X2=367.67, P<0.01).The 2 377 isolated Salmonella strains were classified into 108 serotypes except for oneSalmonella strain that could not be classified.The top four predominant serotypes were 4,5,12:i:-, Salmonella enteritidis,Salmonella stanley and Salmonella typhimurium.Most Salmonella strains were sensitive to imipenem, azithromycin, ceftazidime, cefotaxime and trimethoprim/sulfamethoxazole, but multidrug resistance was common among those strains.Conclusion Salmonella serotypes of 4,5,12:i:-and Salmonella enteritidis are the predominant pathogens causing human Salmonella infections in Guangdong province.Ceftazidime and cefotaximeare are preferred in the treatment of Salmonella infections.Surveillance for drug resistance in Salmonella should be strengthened as multidrug resistant strains have become a serious problem in Guangdong province.
8.Preparation of simulated stool specimens for proficiency testing scheme on the detection of Salmonella and Shigella
Dongmei HE ; Bixia KE ; Hailing TAN ; Bosheng LI ; Honghui ZENG ; Tong YANG ; Changwen KE ; Yuheng LIANG ; Meizhen LIU
Chinese Journal of Microbiology and Immunology 2016;36(7):506-511
Objective To prepare simulated stool specimens for proficiency testing ( PT) by mix-ing lentils with Salmonella, Shigella and Escherichia coli strains and to establish an assessment scheme for the detection of Salmonella and Shigella in clinical samples. Methods Salmonella, Shigella and Escherich-ia coli strains were respectively spiked to lentils in Cary-Blair transport medium to create simulated stool specimens. Various ratios of Escherichia coli to Salmonella strains were spiked to lentils to prepare mixed simulated stool specimens. The accuracy and stability of prepared stool samples for PT were tested in-house. Results of sample detection were collected from participating laboratories for further external quality assess-ment. Results The Escherichia coli and Salmonella strains mixed at ratios of 100 ∶ 1 to 106 ∶ 1 could be ef-ficiently isolated from the media. Enrichment was needed in order to effectively isolate Salmonella strains from the media when the ratios of Escherichia coli to Salmonella strains were 104 ∶ 1 to 106 ∶ 1. Of the16 participating laboratories, 14 laboratories (87. 5%) received a grade of“satisfactory” and the other 2 labo-ratories (12. 5%) received a grade of “mainly satisfactory”. Conclusion The simulated stool specimens and the PT procedures designed in this study were suitable for proficiency testing program on the detection of Salmonella, Shigella and other similar microbes.
9.The Efficacy and Prognostic Factors for Long Pulse Neodymium: Yttrium-Aluminum-Garnet Laser Treatment on Onychomycosis: A Pilot Study.
Sha LU ; Jing ZHANG ; Yuheng LIANG ; Xiqing LI ; Wenying CAI ; Liyan XI
Annals of Dermatology 2016;28(3):406-408
No abstract available.
Neodymium*
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Onychomycosis*
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Pilot Projects*
10.Monitoring of diarrheagenic Escherichia coli infections in patients in Guangdong Province during 2013 to 2016
Dongmei HE ; Yuheng LIANG ; Bixia KE ; Honghui ZENG ; Meizhen LIU ; Hailing TAN ; Bo-Sheng LI ; Tong YANG ; Changwen KE
Chinese Journal of Microbiology and Immunology 2018;38(5):345-353
Objective To investigate the infection status, serotype distribution, drug sensitivity and molecular characteristics of diarrheagenic Escherichia coli (DEC) in patients with diarrhea in Guangdong Province. Methods Fecal samples were collected, cultured and isolated by traditional methods. Suspected Escherichia coli isolates were confirmed by multiplex PCR used for detecting specific virulence genes and bio-chemical methods. Positive strains were serotyped, characterized for drug sensitivity and analyzed by pulsed-field gel electrophoresis ( PFGE). Results The total positive rate of DEC in patients with diarrhea was 6.26%. The positive rates of enteropathogenic Escherichia coli (EPEC), enterotoxigenic Escherichia coli (ETEC), enterohemorrhagic Escherichia coli (EHEC), enteroadherent Escherichia coli (EAEC) and en-teroinvasive Escherichia coli (EIEC) were 2. 47% , 1. 54% , 1. 32% , 0. 62% and 0. 09% , respectively, with infections primarily in children aged 0-<7 years. The total seropositive rate was 52. 54% , with EHEC accounting for 15. 00% . DEC showed high sensitivity to imipenem, ciprofloxacin, ceftazidime and cefo-taxime. The multidrug resistance rate of DEC was 58. 45% , with EPEC being the most serious for multidrug resistance. PFGE results showed that ETEC, EHEC, EPEC and EAEC had a high degree of polymorphism. Conclusion EPEC is the predominant type of DEC circulating in Guangdong Province. Third-generation cephalosporins are the first drugs of choice for treating infections in children. Ciprofloxacin can be used to treat adults. The problem of multiple drug resistance of DEC is severe and efforts to monitor DEC infections and drug resistance should be strengthened.