1.The Imaging Diagnosis of Lipoma of Upper Alimentary Canal
Zhenlong SONG ; Xiang ZHAO ; Jiwei MA
Journal of Practical Radiology 2001;0(10):-
Objective To evaluate the imaging manifestations in diagnosis lipoma of upper alimentary canal.Methods 8 cases with lipoma of upper alimentary canal were performed by barium meal and dual-contrast gastrointestinography(n=8),ultrasound(n=1),computed tomography(n=2)and endoscopy(n=5).Results (1)Barium meal and dual-contrast gastrointestinography could diagnose benign space-occupation of upper alimentary canal.(2)Ultrasound and endoscopy could diagnose benign space-occupation.(3)The density of fat of lesion could well be displayed by computed tomography and contrast-enhenment computed tomography and lipoma could be diagnosed before opertion.Conclusion The combination of gastrointestinography,endoscopy and computed tomography can be regarded as the first procedure for diagnosis lipoma of upper alimentary canal.
2.Cloning, expression and activity identification of human osteoprotegerin-mycobacteria heat shock protein 70 fusion protein
Jing MA ; Wei WANG ; Wenming ZHAO ; Shentao LI ; Mu ZENG ; Zhenlong LIU
Chinese Journal of Tissue Engineering Research 2007;0(15):-
AIM: To solve the two difficulties of bone resorption and inflammation in rheumatoid arthritis, clone the recombinant human osteoprotegerin (OPG) and mycobacteria heat shock protein 70 (HSP70) functional gene,and study the expression and activity of OPG-HSP70 fusion protein in E.coli. METHODS: Experiments were performed at the Laboratory of Department of Immunology, Capital Medical University from May 2006 to September 2007. Complementary DNA encoding full length OPG protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma cell line MG63 and cloned into pGEMT-Easy vectors. Then, using the recombinant plasmid as the template,the DNA encoding the fusion protein OPG-HSP 70 was amplified by PCR,and was inserted into prokaryotic expression vector pET-28a. Construct pET-28a-OPG-HSP 70 was used to transform into competent E.coli. BL21(DE3) which were induced by isopropyl B-D-thiogalactopyranoside (IPTG),and the fusion protein from above E.coli was collected. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blotting were performed to identify OPG-HSP 70 fusion protein. The experiments of osteoclast inhibition and restraining inflammation were used to detect the bioactivity of fusion protein. RESULTS: ①The complementary DNA encoding full length OPG protein was obtained. OPG-HSP 70 fusion gene obtained in this experiment was successfully inserted into pET-28a vector. OPG-HSP70 fusion protein was expressed when transformed into E.coli.BL21(DE3). ②SDS-PAGE indicated that the fusion protein was large expressed at the molecular weight of Mr22 000, but there were no band in the total lysate of bacteria harboring pET-28a-OPG-HSP70 without IPTG induction group. ③Western-blotting indicated that the OPG-HSP70 fusion protein could specifically react with anti-human OPG monoclonal antibodies. ④The osteoclast inhibition test demonstrated that the fusion protein could reduce the number of osteoclast, and had the ability to inhibit bone absorptions in vitro. ⑤The experiment of restraining inflammation showed that the fusion protein could significantly reduce the inflammation of delayed type hypersensitivity (DTH) mice, which explained that HSP70 in fusion protein had the inflammation inhibitory bioactivity. CONCLUSION: OPG-HSP70 fusion protein is expressed in E.coli.BL21(DE3),and function study in vitro illustrates the bioactivity of fusion protein.
3.Establishment and application of a high-throughput screening assay for premature activation of HIV-1 precursors.
Quan ZHANG ; Xiaoyu LI ; Zhenlong LIU ; Pingping JIA ; Xiaolu WEI ; Lixun ZHAO ; Jiandong JIANG ; Shan CEN
Acta Pharmaceutica Sinica 2010;45(2):247-52
Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).
4.Establishment and drug susceptibility test of isoniazid resistant Mycobacterium smegmatis.
Pingping JIA ; Lili ZHAO ; Xiaoyu LI ; Quan ZHANG ; Zhenlong LIU ; Xin WANG ; Liyan YU ; Lixun ZHAO ; Shan CEN
Acta Pharmaceutica Sinica 2011;46(4):377-82
With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.
5.Tendency analysis of incidence and mortality of mechanical injuries among Chinese residents from 1990 to 2019
Tao LIU ; Zhao YANG ; Fangguo LI ; Yue LI ; Xin LIU ; Zhenlong WU ; Chunxia CAO
Chinese Critical Care Medicine 2022;34(10):1082-1087
Objective:To analyze the tendency of incidence and mortality of mechanical injuries among Chinese residents from 1990 through 2019 and to estimate the age-period-cohort effect.Methods:Based on the Global Burden of Disease (GBD) 2019 database, the incidence and mortality data of mechanical injuries among Chinese residents from 1990 to 2019 were extracted. The trends of age-standardized incidence rate (ASIR) and age-standardized death rate (ASDR) of mechanical injuries among Chinese residents by gender was analyzed using the joinpoint regression model, and the annual percent change (APC) and average annual percent change (AAPC) were calculated. The age-period-cohort model was used to quantitatively assess the effects of age, period, and cohort on ASIR and ASDR for mechanical injuries.Results:① Overall tendency: from 1990 to 2019, the ASIR of mechanical injuries showed an increasing trend (540.95/100 000 in 1990 vs. 815.34/100 000 in 2019), and the ASDR first increased slightly and then decreased (2.62/100 000 in 1990 vs. 2.87/100 000 in 2005 vs. 1.77/100 000 in 2019) among Chinese residents. During the observation period, ASIR and ASDR for mechanical injuries of male were higher than female. ② Joinpoint regression model analysis showed that the ASIR of mechanical injuries had a fluctuating trend of increasing first and then decreasing and then rising rapidly among Chinese residents from 1990 to 2019 (AAPC = 1.42%, t = 9.59, P < 0.001). The ASIR of the Chinese male showed a slight decrease and then continued to increase (AAPC = 1.47%, t = 8.72, P < 0.001), while the ASIR of the Chinese female showed a rapid rising at first, then rapidly declining and then rising again (AAPC = 1.31%, t = 12.11, P < 0.001). From 1990 to 2019, the ASDR of mechanical injuries showed a fluctuating downward trend of first decreasing, then increasing, and then rapidly decreasing among Chinese residents (AAPC = -1.39%, t = -6.72, P < 0.001). The decrease rate of ASDR among male was as same as that among all population (AAPC = -1.44%, t = -7.29, P < 0.001), but the decrease rate of ASDR in female was relatively slow (AAPC = -1.08%, t = -4.54, P < 0.001). ③ Age-period-cohort model analysis showed that, with the increase of age, the risk of mechanical injuries among the overall population, male and female in China increased, then decreased, and then increased rapidly. The first small peak was at the age of 45-49 years old in male and 65-69 years old in female. The overall death risk showed an increasing trend with age, with a slowly increasing trend before 75 years old, and a sudden increase after 75 years old. The peak age of death risk was between 90 and 94 years. In terms of period effect, the risk of mechanical injuries showed a gradually increasing trend with time among the overall population, male, and female in China, and the risk of death showed a trend of decreasing first and then rapidly increasing and then decreasing. In terms of cohort effect, the risk of mechanical injuries among the overall population, male, and female in China showed a gradual upward trend with the increase in the birth year, and the risk of death showed an M-shaped trend. Conclusions:From 1990 to 2019, the incidence of mechanical injuries showed an increasing trend, and the mortality increased first and then decreased. Although the disease burden has improved, it is still high. More attention needs to be paid to the prevention and control of mechanical injuries, especially in the young population.
6.Effect of mean arterial pressure on arterial to end-tidal CO2 partial pressure difference dur-ing one-lung ventilation
Zhaoyan ZENG ; Jing YE ; Zhenlong ZHAO ; Zaisheng QIN
Journal of Southern Medical University 2014;(12):1834-1837
Objective To study the effect of different mean arterial pressures (MAP) on arterial to end-tidal CO2 partial pressure difference [(Pa-et)CO2] and the intrapulmonary shunt (Qs/Qt) in patients undergoing thoracic surgery during one-lung ventilation (OLV). Methods Forty-two patients undergoing right-sided pulmonary lobectomy were allocated into group A with fluctuation of MAP (at 20 min after OLV) within ± 10%of the baseline (n=22) and group B with lowered MAP by 30%of the baseline (n=20). Arterial and venous blood gas analyses were recorded to calculate [(Pa-et)CO2] and Qs/Qt at 20 min after induction with two-lung ventilation (T1), 20 min after OLV (T2), 30 min after recovery of normal blood pressure (T3), and 20 min after recovery of two-lung ventilation (T4). Results PetCO2 and PaCO2 were well correlated during two-lung ventilation and OLV in group A (P<0.05). In group B, [(Pa-et)CO2] at T2 was significantly higher than that in group A, but PetCO2 was still correlated with PaCO2 (P<0.05). Qs/Qt increased more obviously in group B than in group A in T2 (P<0.05). Bo obvious correlation was found between the [(Pa-et)CO2] and Qs/Qt during OLV. Conclusion PetCO2 reflects the dynamic changes of PaCO2 under normal blood pressure during OLV. In the hypotension period, when [(Pa-et)CO2] increases and the correlation coefficient between PetCO2 and PaCO2 lowers, PetCO2 may not accurately reflect the changes of PaCO2 and blood gas analysis is warranted.
7.Effect of mean arterial pressure on arterial to end-tidal CO2 partial pressure difference dur-ing one-lung ventilation
Zhaoyan ZENG ; Jing YE ; Zhenlong ZHAO ; Zaisheng QIN
Journal of Southern Medical University 2014;(12):1834-1837
Objective To study the effect of different mean arterial pressures (MAP) on arterial to end-tidal CO2 partial pressure difference [(Pa-et)CO2] and the intrapulmonary shunt (Qs/Qt) in patients undergoing thoracic surgery during one-lung ventilation (OLV). Methods Forty-two patients undergoing right-sided pulmonary lobectomy were allocated into group A with fluctuation of MAP (at 20 min after OLV) within ± 10%of the baseline (n=22) and group B with lowered MAP by 30%of the baseline (n=20). Arterial and venous blood gas analyses were recorded to calculate [(Pa-et)CO2] and Qs/Qt at 20 min after induction with two-lung ventilation (T1), 20 min after OLV (T2), 30 min after recovery of normal blood pressure (T3), and 20 min after recovery of two-lung ventilation (T4). Results PetCO2 and PaCO2 were well correlated during two-lung ventilation and OLV in group A (P<0.05). In group B, [(Pa-et)CO2] at T2 was significantly higher than that in group A, but PetCO2 was still correlated with PaCO2 (P<0.05). Qs/Qt increased more obviously in group B than in group A in T2 (P<0.05). Bo obvious correlation was found between the [(Pa-et)CO2] and Qs/Qt during OLV. Conclusion PetCO2 reflects the dynamic changes of PaCO2 under normal blood pressure during OLV. In the hypotension period, when [(Pa-et)CO2] increases and the correlation coefficient between PetCO2 and PaCO2 lowers, PetCO2 may not accurately reflect the changes of PaCO2 and blood gas analysis is warranted.
8.Difference of Graft Maturity of Anterior Cruciate Ligament Reconstruction between Oval-Shaped Tunnel Technique and Round-Shaped Tunnel Technique
Jiahao ZHANG ; Zhenlong LIU ; Xiaoqing HU ; Fengyuan ZHAO ; Yong MA ; Haijun WANG ; Yingfang AO
Chinese Journal of Sports Medicine 2018;37(2):104-109
Objective To compare the graft maturity of the anterior cruciate ligament reconstruction using the oval-shaped tunnel technique and round-shaped tunnel technique by signal/noise quotient (SNQ) of MRI postoperatively.Methods Forty patients diagnosed as the anterior cruciate ligament rupture between August 2015 and June 2016 were included according to the inclusion and exclusion criteria.Patients were randomized into a round-shaped group and an oval-shaped group,undergoing traditional round-shaped tunnel and oval-shaped tunnel reconstruction of the anterior cruciate ligament respectively.One year postoperatively,the MRI was conducted,and three intra-articular regions of interest (ROI) were selected to compare the graft maturity by calculating SNQ.Results None of the forty patients experienced complications of bone tunnel blowout,graft getting through difficulties and neurological or vascular injuries.SNQ of the round-shaped group were 3.72 ± 2.29,significantly higher than that of the oval-shaped group(P<0.001).Moreover,SNQ of proximal ROI and distal ROI of ovalshaped group were 1.97 ± 1.30 and 2.76 ± 1.75,significantly lower than the round-shaped group with proximal site of 3.53 ± 2.11(P=0.008) and distal site of 4.46 ± 2.28(P=0.012).Conclusion Comparing MRI signal intensity one year after the treatment,we have found the graft SNQ after oval-shaped tunnel reconstruction was lower than the round-shaped reconstruction,with better graft maturity.
9.MAPK4 Accelerates Progression of Cervical Squamous Cell Carcinoma by Positively Regulating SLC3A2 Expression
Jing YU ; Lu DENG ; Yuting ZHAO ; Zhenlong YUAN ; Lingying WU
Cancer Research on Prevention and Treatment 2024;51(10):803-812
Objective To explore the role of MAPK4 in cervical squamous cell carcinoma(CSCC)for the identification of candidate prognostic prediction biomarkers and molecular therapeutic targets.Methods The TCGA cohort was subjected to Kaplan-Meier survival analysis.Immunohistochemistry experiments were conducted on clinical samples to explore the correlation between MAPK4 and patient prognosis.A nomogram was constructed based on MAPK4 mRNA levels.Western blot,CCK-8,colony formation,and Transwell cell function experiments were performed to clarify the abnormal expression and role of MAPK4 in CSCC.DIA proteome sequencing was used to identify effector molecules regulated by MAPK4.Combined with the above cell function experiments,the knockdown of MAPK4 and the overexpression of effector molecules revealed that MAPK4 regulated effector molecules to mediate tumor progression.Results CSCC patients with elevated MAPK4 mRNA levels and high protein expression have a worse prognosis.The constructed nomogram based on MAPK4 can accurately predict the 1-,3-,and 5-year survival rates of patients.Compared with that in normal cervical tissues,MAPK4 protein expre-ssion was up-regulated in tumors.MAP-K4 knockdown substantially inhibited the proliferation,colony formation,mig-ration,and invasion of CSCC ME180 and SiHa cells.SLC3A2 is a downs-tream effector molecule of MAPK4.Overexpression SLC3A2 can weaken the inhibitory effect of MAPK4 knockdown on cell proliferation,colony formation,migration,and invasion.Conclusion MAPK4 is a candidate prognostic biomarker for patients with CSCC.MAPK4 positively regulates SLC3A2 protein expression and accelerates tumor progression,making it a potential molecular therapeutic target for patients with CSCC.