1.Study on silence of survivin gene in cancer cells A549, Hela S3 and K562 by small interfering RNA and its sequence screening
Mengze HU ; Xiaodong SHI ; Tianyou WANG ; Rong LIU ; Litian XUAN ; Yanfei CHEN
Chinese Journal of Applied Clinical Pediatrics 2014;29(3):194-198
Objective To investigate the biological response of survivin siRNA in A549 human lung cancer cells and Hela S3 human cervical cancer cells as well as K562 human erythroleukemia cells,in order to screen the working sequences of survivin siRNA.Methods Three sequences of survivin-targeted siRNA were designed and synthetized,and human cancer lines of A549,Hela S3,K562 were transfected with Hiperfect liposome entrapped survivin siRNA,respectively.The expression of survivin mRNA was detected by means of real-time polymerase chain reaction (RT-PCR) with SYBR Green Ⅰ.Cell proliferation was detected by way of WST-8 cell count kit at 48 hours and 72 hours after transfection.Results The expression of survivin mRNA in all 3 cancer cells studied was significantly inhibited by all siRNA at 48 hours and 72 hours after transfection,gene expression inhibition ratio were 57.47%-88.53% after transfection 48 hours and were 69.94%-95.03% after transfection 72 hours.Cell proliferation was also significantly inhibited 48 hours and 72 hours after transfection,cell multiplication inhibition ratio were 27.88%-47.36% after transfeotion 48 hours and were 42.59%-57.29% after transfeciton 72 hours.Sequence 1 had the most inhibition efficacy on survivin gene expression and proli-feration in tumor cells.Inhibition rate of the three tumor cell gene expression of survivin at 48 hours are above 75%,72 h all over 90%,48 hours of cell proliferation inhibition rate are above 40%,72 hours of more than 50%.Conclusions The chemo-synthesized siRNAs can significantly down-regulate survivin mRNA expression in cancer cell studies.Survivin siRNA is capable of inhibiting the proliferation of tumor cell in vitro and it might be a new strategy for tumor-targeted therapy.Sequence 1 is the most efficacious working survivin siR-NA in the study.
2.Case 04 (2024): Two cases of neonatal limb arterial thrombosis with thrombolytic therapy
Mengze SUN ; Ying ZHANG ; Laishuan WANG ; Zheng CHEN ; Hua WANG ; Xin DING ; Qiushi WANG ; Haidi HU ; Ana HOU
Chinese Journal of Perinatal Medicine 2024;27(8):688-694
This article reported two cases of axillary artery thrombosis in extremely low/very low birth weight infants following the placement of a local arterial catheter, who hospitalized in Shengjing Hospital of China Medical Universityin in April 2023 and August 2022, respectively. Case 1: Before surgery for necrotizing enterocolitis, an arterial catheter was placed in the left axilla of the infant. On the same day, the infant developed cyanosis of the left upper limb and weakened radial artery pulse. Ultrasound examination confirmed the presence of left axillary artery thrombosis. Despite subcutaneous injection of low molecular weight heparin (LMWH) and plasma infusion, there was no improvement in blood circulation. The infant also exhibited reduced movement in the left upper limb and loss of radial artery pulse. Thrombolytic therapy with recombinant tissue-type plasminogen activator was administered. Six hours after the treatment, the radial artery pulse became palpable. Thrombolysis was then terminated, and anticoagulation with LMWH was supplied for two weeks. At one year and eight months of age, the infant had a weaker left-hand grip strength compared to the right, but the overall functionality was largely preserved. Case 2: The infant developed late-onset sepsis at 17 days old and had an arterial catheter placed in the axilla. Pale left upper limb was observed in the following day, and the brachial and radial artery pulses were absent. Vascular ultrasound indicated the presence of left axillary artery thrombosis. Anticoagulation therapy with subcutaneous injection of LMWH was provided, along with thrombolysis using urokinase. On the sixth day after thrombolysis, an ultrasound examination showed no thrombus-like echoes. At one year and eight months of age, the development and movement of the affected upper limb became normal.
3.Analysis on drug resistance in people infected with HIV-1 CRF55_01B before antiviral therapy in China
Shan ZHENG ; Chang SONG ; Mengze GAN ; Qingyu HAN ; Jing HU ; Aobo DONG ; Lingjie LIAO ; Yi FENG ; Yuhua RUAN ; Hui XING
Chinese Journal of Experimental and Clinical Virology 2021;35(2):194-198
Objective:To explore the resistance to protease inhibitors (PI), nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) and integrase inhibitors (INSTIs) of HIV-1 CRF55_01B and the transmission of drug-resistant strains among HIV-1 CRF55_01B infected patients before antiviral treatment in China.Methods:HIV-1 RNA was extracted from plasma samples of the patients infected with CRF55_01B in the national surveillance of HIV drug resistance before antiviral treatment in 2018. A 1 056 bp gene fragment of protease/reverse transcriptase (PR/RT) region and an 846 bp gene fragment of integrase (IN) region were obtained and sequenced. Drug resistance was analyzed by using all drugs included in the Stanford University HIV db Program, HIV-1 molecular network analysis was performed with software HIV-TRACE and polymorphism mutations of CRF55_01B integrase gene region were analyzed.Results:A total of 178 samples from 26 provinces, municipalities and autonomous regions in China were analyzed, and 170 sequences of CRF55_01B PR/RT region and 170 sequences of IN region of corresponding samples were obtained. The drug resistance rate was 15.3% (26/170). The drug resistance rates of PIs, NRTIs, NNRTIs and INSTIs were 1.2% (2/170), 1.2% (2/170), 15.3% (26/170), 0.6% (1/170), respectively. The level of drug resistance was mostly low. NNRTIs drug resistance mutations were mainly V179D/E co-appeared with other mutations, and 84.1% (143/170) of the infected patients carrying V179D/E alone showed potential drug resistance. INSTIs drug resistance mutation was G163R, and showed low resistance to EVG and RAL. The molecular network access rate was 30.0%(51/170)according to the 0.9% gene distance threshold. The resistant strains were transmitted between men with homosexual transmission and heterosexually transmitted people, and both carried resistant mutations E138G and V179E. In the integrase region, CRF55_01B and CRF01_AE and B subtypes showed high mutation frequency difference in 5 sites (T215A、G134N、I135V, K136R and L101I/V).Conclusions:Before antiviral treatment, CRF55_01B infected patients in China had a high resistance to NNRTIs. Strains carrying both E138G and V179E resistance mutations were transmitting in clusters. The prevalence of CRF55_01B integrase inhibitor resistant strains is low, but some genetic polymorphisms with high mutation rate in the integrase gene region have potential influence on drug sensitivity. The influence of drug resistance of new recombinant strains on antiviral therapy in China needs to be further monitored and analyzed.