1.Construction of cell line with overexpression of rat serotonin1A receptor
Zhihua LI ; Bojuan WU ; Wanjun SONG ; Yi DANG ; Xiaoyong QI
Chinese Journal of Anesthesiology 2011;31(1):74-77
Objective To establish a cell line with overexpression of rat serotonin1A receptor (5-HT1AR).Methods Human neuroblastoma cells-SH-SY5Y were donated by cancer institute attached to the 4 th Affiliated Hospital, Hebei Medical University. Total RNA was extracted from brain tissues of male SD rats and rat 5-HT1A R was obtained by RT-PCR. Plasmid pc-DNA3. 1/hisC containing the rat 5-HT1AR (pc-DNA3.1/hisC-Rat-5-HT1AR)was constructed and transfected into SH-SY5Y cells. The transfected cells were isolated by G418 selection and SH-SY5Y-Rat-5-HT1A R cells were obtained. Expression of 5-HT1A R was detected by Western blot analysis. Cell viability was evaluated by MTT assay. SH-SY5Y-Rat-5-HT1AR cells were further observed for 5-HT1AR by immuno-fluorescence staining. Results Plasmid pc-DNA3. 1/hisC-Rat-5-HT1AR was successfully constructed by linking Rat-5-HT1A R with pc-DNA3.1/hisC and transfected into SH-SY5Y. The SH-SY5Y-Rat-5-HT1A R cells were more slender than SH-SY5Y cells with less and longer processes. MTT showed that the viability of SH-SY5Y-Rat-5-HT1A R cells was much lower than SH-SY5Y. Rat 5-HT1A R was expressed efficiently on the membrane of SH-SY5Y-Rat-5-HT1A R cells. Conclusion A cell line with overexpress of rat 5-HT1A R is successfully established.
2.Effects of electroacupuncture on cognitive function and learning and memory of mice exposed to ionizing radiation
Xin WU ; Wanjun ZHENG ; Lin LI ; Jianfeng GAO
Chinese Journal of Radiological Medicine and Protection 2016;36(2):94-99
Objective To investigate the effects of electroacupunctures at Baihui,Fengfu,and Shenshu on the cognitive function and learning memory of mice exposure with ionizing radiation.Methods C57BL/6J (30-day old) mice were randomly divided into l0 groups consisting of the control group,X-ray exposure groups (4,8 and 16 Gy),electroacupuncture-nonacupiont (E-nonA) groups (4,8 and 16 Gy) and electroacupuncture-acupiont (EA) groups (4,8 Gy and 16 Gy),with 10 mice assigned to each group.The mice were irradiated (IR) on the left cerebral hemisphere with doses of 4,8 and 16 Gy.Some mice were electroacupunctured for 3 courses of treatment.Using weight as a measurable parameter,the open-field test and Morris water maze test were used to evaluate the curative effects.Results Compared with the control group,the weights of mice in the exposure groups (8 and 16 Gy) significantly decreased (t =2.917,9.650,3.043,2.882,P < 0.05) at 7 and 14 d,while the weights of the E-nonA (8 and 16 Gy) and EA groups (8 and 16 Gy) were not significantly altered.Compared to the model groups (8 and 16 Gy),the weights of EA groups (8 and 16 Gy) significantly increased (t =6.602,5.409,19.660,2.975,P < 0.05).In the open-field test,the crossing times and the rearing time of exposure model groups (8 and 16 Gy) significantly decreased compared to the control group (t =2.183,2.119,2.369,2.231,P < 0.05).Compared with model groups (8 and 16 Gy) and E-nonA groups (8 and 16 Gy),the crossing times and rearing time of the EA groups were significantly increased (compared with model:t =8.914,2.727,4.325,4.178,P < 0.05;compared with E-nonA:t =4.030,3.014,2.150,4.771,P <0.05).In the Morris water maze,compared with the control,the latent period of model group (4 Gy) was significantly increased and the crossing times significantly decreased (t =4.544,3.422,P < 0.05);compared to model and E-nonA groups (4 Gy),the latent period of EA group significantly increased (compared with model:t =2.877,P < 0.05;compared with E-nonA:t =3.001,P < 0.05).At day 2 and 3,the latent period of model groups (8 and 16 Gy) significantly increased (t =2.544,3.706,3.796,7.934,P < 0.05) and the crossing times (t =2.120,2.393,P < 0.05) significantly decreased,resident time in target area of model group (8 Gy) was significantly decreased (t =2.543,P < 0.05) compared to the control group.Compared with model and E-nonA groups (8 and 16 Gy),the latent period of EA gourps significantly decreased (compared with model:t =5.404,7.869,4.104,15.590,P < 0.05;compared with E-nonA:t =2.938,7.955,6.566,10.350,P < 0.05) while the through times and resident time in target area significantly increased in the EA group (8 Gy) (compared with model:t =2.673,2.613,P < 0.05;compared with E-nonA:t =3.345,2.179,P < 0.05).Conclusions Irradiation with a dose higher than 8 Gy could induce mice brain injury.Electroacupuncture can alleviate the radiation damage of cognitive function as well as learning and memory functions in mice.
3.Experimental study on different courses of electro-acupuncture treatment on behavior of model mice with radiographic exposure
Xin WU ; Wanjun ZHENG ; Lin LI ; Jianfeng GAO
Chinese Journal of Radiation Oncology 2016;25(6):634-638
Objective To explore the effects of different courses of electro-acupuncture at Baihui,Fengfu,and Shenshu of both sides on the behavior of mice exposed to radiation.Methods Fifty C57BL/6Jmice 30 days old were randomly and equally divided into control group,model group (exposure to radiation of 8 Gy),electro-acupuncture group 1 (1 week of treatment),electro-acupuncture group 2 (2 weeks of treatment),and electro-acupuncture group 3 (3 weeks of treatment).The mouse model of radiation-induced brain injury was established by exposure to radiation of 8 Gy and treated with electro-acupuncture.The open field test and Morris water maze test were used to evaluate the cognitive function and spatial learning and memory,respectively.Group t-test difference.Results In the open field test,the model group had significantly fewer vertical or horizontal movements than the control group (P=0.032,0.029,0.028),while the electro-acupuncture groups 2 and 3 had significantly more vertical or horizontal movements than the model group (P=0.001,0.017,0.000).In the Morris water maze test,compared with the control group,the model group had significantly longer latency,a significantly lower frequency of entry into the goal box,and significantly shortened residence time,indicating substantially worse performance of spatial learning and memory (P=0.023,0.001,0.001).Compared with the model group,the electro-acupuncture groups 2 and 3 had significantly shortened latency,a significantly higher frequency of entry into the goal box,and significantly longer residence time (P=0.046,0.017,0.049,0.007,0.031,0.009).Conclusions Different courses of electro-acupuncture treatment at Baihui,Fengfu,and Shenshu of both sides can substantially improve the cognitive function and spatial learning and memory in mice exposed to radiation.Particularly,2 and 3 weeks of electro-acupuncture treatment give the best outcomes.
4.Influencing factors of lateral lymph node metastasis in papillary thyroid carcinoma
Liang WANG ; Changhua WU ; Shujuan ZOU ; Wanjun CHEN
Journal of International Oncology 2016;43(3):207-209
Papillary thyroid carcinoma is the most common pathology type of thyroid carcinoma with the following biological characteristics:low malignancy,slow progress,longer duration,good prognosis,but it is prone to lymph node metastasis.Lateral lymph node metastasis relates with patients' age,sex,tumor size,tumor location,capsular invasion/extra-thyroidal extension,muli-focality and the number of positive central lymph nodes.Understanding the influencing factors of the lateral lymph node metastasis has important clinical significance for determining the lymph node dissection scope and the indications for the patients with thyroid carcinoma.
5.Label-Free Detection of Immunoglobulin G Based On Silicon-on-Insulator Microring Resonator Analysis System Integrated with Microfluidic Channels
Yingying GUO ; Shangquan WU ; Li JIN ; Wanjun WANG ; Qingchuan ZHANG
Chinese Journal of Analytical Chemistry 2017;45(4):483-488
The microfluidic channels integrated with microring resonator were designed. The salt coalescence on chip surface caused by liquid volatilization in open environment was avoided and only 30 μL of reaction solution was consumed. These channels significantly reduced the experiment cost. The design, fabrication and characterization of a highly sensitive and label-free silicon-on-insulator (SOI) microring optical resonator integrated with the microfluidic channels were demonstrated. The radius of the microring was 5 μm and the straight waveguide with a width of 450 nm was employed in the microring resonator. The microring resonator device had many advantages such as high sensitivity, label-free and real-time detection. Using different concentrations of ethanol solution with known refractive indices, the refractive index detection sensitivity was 76.09 nm/RIU and the volume refractive index detection limit was 5.25×10Symbolm_4 RIU. We also demonstrated the label-free quantitative specific detections of human immunoglobulin G (IgG) solutions using an antibody-modified microring resonator by measuring the resonance wavelength shift resulting from refraction index changes causing by the immobilization of antibodies and specific recognition between antibodies and antigens, respectively. The results showed that the microring optical resonator could real-time monitor the reaction between biological molecules, the resonator could be used in the quantitative detection and biological sensing.
6.Effect of silicon dioxide exposure on airway surface microenvironment and NEK7/NLPR3 inflammasome in rats
Wenlu HANG ; Qi WU ; Wanjun LI ; Yun BO ; Xianmei ZHOU
Journal of Preventive Medicine 2023;35(2):180-184
Objective:
To examine the effect of SiO2 exposure on the airway surface microenvironment and NIMA-related kinase 7 (NEK7)/nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasome in rats.
Methods:
Twenty-four specific pathogen-free male rats of the SD strain were randomly divided into the control group and the model group, of 12 rats in each group. Rats in the model group were given SiO2 suspensions through disposable tracheal intubation perfusion to model silicosis in rats, while rats in the control group was perfused with the same amount of physiological saline. The pH value and glucose level were measured in the rat bronchoalveolar lavage fluid (BALF) 14 and 28 days after modeling. Lung tissues were stained with HE and Masson and the distribution of inflammatory cells and the deposition of pulmonary interstitial collagens were observed in lung tissues under a light microscope. The expression of transforming growth factor β1 (TGF-β1), collagen type Ⅰ(ColⅠ), collagen type Ⅲ (Col Ⅲ), interleukin-1β (IL-1β), NLRP3, N-terminal domain of Gasdermin D (GSDMD-NT), caspase-1, and NEK7 was quantified in lung specimens using immunohistochemistry.
Results:
Lower pH values were measured in rat BALF in the model group than in the control group 14 [(6.38±0.05) vs. (6.68±0.08), P<0.05] and 28 days after modeling [(6.63±0.14) vs. (6.86±0.05), P<0.05], while higher glucose levels were seen in the model group than in the control group 14 [(0.39±0.06) vs. (0.31±0.04) mg/dL, P<0.05] and 28 days after modeling [(0.39±0.08) vs. (0.31±0.06) mg/dL, P<0.05]. HE and Masson staining showed mild to moderate alveolitis and pulmonary fibrosis in rats 14 days post-exposure to SiO2, and showed moderate to severe alveolitis and pulmonary fibrosis 28 days post-exposure. Immunohistochemistry detected higher TGF-β1, ColⅠ, Col Ⅲ, IL-1β, NLRP3, GSDMD-NT, caspase-1 and NEK7 expression in rat lung tissues in the model group than in the control group (all P<0.05).
Conclusions
SiO2 exposure may cause changes in rat airway surface microenvironment, including BALF acidification and elevated glucose. Pyroptosis induced by activation of NEK7-associated NLRP3 inflammasome may be an important mechanism of pulmonary fibrosis caused by silicosis.
7.The Application of UART-Wi-Fi Module in the Physiological Signal Wireless Measurement System
Zeli GAO ; Jie WU ; Wanjun YANG ; Jianli ZHOU ; Zhiren WU ; Wei JIANG
Journal of Kunming Medical University 2013;(10):27-30
Objective To explore a new way of physiological signal wireless measurement to popularize the physiological signal wireless measurement. Methods We added a UART-Wi-Fi module between the signal detecting module and the PC, the physiological signal measurement system transmitted the signal data collected by the single chip computer to the UART-Wi-Fi module through the serial interface RS-232C. Then the UART-Wi-Fi module sent the signal data out to the Wi-Fi wireless network. The PC received the signal data from the Wi-Fi wireless network and processed the signal data, then output the results. Result Through the UART-Wi-Fi module, the communication between PC and the signal detecting module was converted from wired communication into wireless communication successfully. Conclusion As a result of any computer can be used as a physiological signal receiving and processing terminal equipment, the use of the UART-Wi-Fi module can help achieve popularization of physiological signal wireless measurement.
8.Clinical analysis of 44 cases with malignant transformation of ovarian mature cystic teratoma
Jusheng AN ; Lingying WU ; Xiaoguang LI ; Rong ZHANG ; Yan SONG ; Shaokang MA ; Liying LIU ; Wanjun HONG
Chinese Journal of Obstetrics and Gynecology 2013;(2):123-128
Objective To analyze the clinicopathologic characteristics,treatment and prognostic factors in malignant transformation of mature cystic teratoma(MCT)of ovary.Methods The clinical data of 44 patients with MCT from January 1961 to June 2009 were reviewed.Results The median age of the 44 patients was 48 years(range,16-84 years).Mean tumor size was(16 ±6)cm.Thirty-two cases were diagnosed squamous cell carcinoma(73%,32/44),and 5 of them with the elevated level of serumal squamous cell antigen(SCC-Ag).Three of 37 cases(8%,3/37)were identified with malignant transformation in image examinations.Rapid frozen section examination and multiple-location biopsy were performed in 8 cases,and 5 of them were detected with malignant diseases.Twenty-two patients with disease confined within the unilateral ovary(10 with intact capsule,and 12 with ruptured capsule).Diseases extended extra ovaries in the others 22 patients.The median cumulative overall survivals were 126 and 10 months,respectively.The difference between the two groups was significant(P < 0.01).Twenty-seven patients had no residual tumor after primary surgery.The median cumulative overall survivals between the patients with and without residual tumor were 10 and 84 months respectively,and there were significant difference between two groups(P < 0.01).Seven selected patients with malignant disease confined within unilateral ovary underwent fertility-sparing surgery,and 2 cases of them had successful pregnancies and delivery,while other 4 cases with ruptured capsule recurred.Conclusions The most common pathology type of malignant transformation in mature cystic teratoma of the ovary is squamous cell carcinoma.Comprehensive pre-operation image examination and tumor marker level detection might be of great help in diagnosis.Tumor extension extraovary and residual tumor after surgery are the most significant poor prognostic factors.Early stage patient with ruptured capsule should be very discreet to choose fertility-sparing surgery.
9.The investigation of risk factors of post-polypectomy emergency hemorrhage
Yadi ZHANG ; Wensheng PAN ; Hong SHEN ; Wanjun ZHANG ; Liangqin WU ; Wen HU ; Yimiao ZHU ; Xiang XU
Chinese Journal of Emergency Medicine 2012;21(1):74-78
Objective To explore the risk factors in post-polypectomy hemorrhage in rectum and to discuss the appropriate interventions.Methods A total of 313 patients with 373 polypi were included in this study. The clinical data were analyzed by SPSS 16 software. Results There were 313 patients with colorectal polypus curatively resected and 373 polypi in total.There were 11 (3.5%) patients subjected to post-polypectomy hemorrhage in rectum.Regression analysis showed that the independent risk factor of postpolypectomy hemorrhage in rectum was the hypertension of patients (P < 0.01 ) and this hemorrhage had no significant correlations with patientg'ender,age,size of polypus,pathological characteristics and the methods of polypectomy.Conclusions Hypertension of patients is an independent risk factor in post-polypectomy hemorrhage.
10.Study on Simultaneous Determination Method of Zolpidem,Dexzopiclone and Zaleplon in Human Plasma by UPLC-MS/MS
Xin WEI ; Jing AN ; Yin WU ; Zhanjun DONG ; Liju FAN ; Wanjun BAI
China Pharmacy 2016;27(23):3194-3196,3197
OBJECTIVE:To develop a method for rapid,accurate and simultaneous determination of dexzopiclone,zolpidam and zaleplon in human plasma. METHODS:After the plasma sample was processed by liquid-liquid extraction,UPLC-MS/MS was established to detect plasma sample with carbamazepine as internal standard. The separation was performed on a Waters ACQUITY UPLC HSS T3 column with mobile phase composed of 0.1% ammonium solution-acetonitrile (gradient elution) at flow rate of 0.2 ml/min. The column temperature was set at 40 ℃,and sample size was 5 μl. The detection was performed by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. The mass transition ion-pairs were as follows:m/z 308.2→263.1(zolpidem),m/z 389.3→245.0(dexzopiclone),m/z 306.2→236.1(zaleplon),m/z 237.3→151.2(internal standard). RE-SULTS:The linear range of zolpidem,dexzopiclone and zaleplon were 0.02-20.00,0.50-20.00,0.02-20.00 ng/ml,respectively (r=0.990 1,0.996 8,0.991 7). LLOQ of them were 0.02,0.50,0.02 ng/ml,and detection limit were 0.01,0.20,0.01 ng/ml. RS-Ds of intra-day and inter-day were all lower than 15% ;extraction recovery were 88.9% -106.5% ;matrix effect were 94.8%-106.3%. CONCLUSIONS:The method is simple,rapid,sensitive and specific,and it can be used for simultaneous deter-mination of zaleplon,zolpidem and dexzopiclone in human plasma.