1.Detection of expression of mouse MN/CA9 gene with reverse transcriptase-polymerase chain reaction
Yinping WANG ; Guimin WANG ; Shiquan XUE ; Yabin ZOU
Journal of Jilin University(Medicine Edition) 2006;0(05):-
Objective To clone and analyze the MN/CA9 gene sequence in ICR mice and detect the expressions of MN/CA9 gene in various tissues of ICR mice.Methods The fresh tissues of small intesine, uterrus, skin, musle, liver, pancreas, heart, lung, thymus, spleen, kidney, ovary, stomach ,urinary bladder from ICR mice were obtained , the total RNA was extracted by GIT method, the 1st strand and 2nd strand of cDNA were synthesized, the EcoRⅠ adapters were lingated,the EcoRⅠ ends were phosphorylated, digested with XhoⅠ ,cDNA was ligated into the ZAP expression vector, packaged, planted, screened.The expressions of MN/CA 9 gene in various tissues in mice were detected by RT-PCR.Results A fragment of human MN/CA9 gene was used as probe, and 1.47?10~3 clones were screened with radioactive isotopic ~ 32 P labeled probe, after hybridizations, one positive signal of cDNA was detected and the complete nucleotide sequence contained 1 671 bp was determined (GenBank:AB086322), The nucleotide similarity between mouse and human cDNA (GenBank:Z54349) was 69.1%.The MN/CA9 gene detected by RT-PCR assay (primer: P521-P1193) strongly expressed in small intesine,uterus, musle, pancreas, heart, lung, thymus, spleen, kidney, ovary, stomach,and urinary bladder,meanwhile did not express in skin and liver. Conclusion The expressions of MN/CA9 gene in some tissues of ICR mice are similar to that of human, it can be used to further functional analysis of MN/CA9 gene.
2.Effect of Tanshinonne ⅡA preconditioning on nuclear factor kappa B signaling pathway in N9 cells subjected to oxygen-glucose deprivation and restoration
Shiquan WANG ; Tingting GU ; Jing HAO ; Yufei WU ; Xue YANG ; Qianzi YANG
Chinese Journal of Anesthesiology 2019;39(4):486-489
Objective To evaluate the effect of TanshinonneⅡA preconditioning on nuclear factor kappa B ( NF-κB ) signaling pathway in N9 cells subjected to oxygen-glucose deprivation and restoration ( OGD∕R) . Methods N9 cells cultured in vitro were divided into 4 groups ( n=16 each) by using a ran-dom number table method: control group ( C group) , OGD∕R group, vehicle group ( V group) and Tanshi-none ⅡA preconditioning group ( TP group) . N9 cells were subjected to O2-glucose deprivation for 4 h fol-lowed by restoration of O2-glucose supply for 24 h to establish the OGD∕R model. Group C receive no treat-ment. In group TP, N9 cells were cultured in medium containing Tanshinone ⅡA ( final concentration 5 mg∕L) for 24 h before the model was established. Dimethyl sulfoxide was added to the culture medium in group V. At the 24 h of reoxygenation, the level of lactate dehydrogenase ( LDH) and nitric oxide ( NO) in culture medium was detected by spectrophotometry, cell apoptosis was assessed by using TUNEL, and the expression of NF-κB in nucleus was detected by Western blot. The apoptosis rate was calculated. Results Compared with group C, the levels of LDH, IL-β, TNF-α and NO in culture medium and apoptosis rate were significantly increased, and the expression of NF-κB in nucleus was up-regulated in OGD∕R, V and TP groups ( P<0. 05) . Compared with group OGD∕R, the levels of LDH, IL-β, TNF-αand NO in culture me-dium and apoptosis rate were significantly decreased, and the expression of NF-κB in nucleus was down-reg-ulated in group TP ( P<0. 05) , and no significant change was found in the parameters mentioned above in group V ( P>0. 05 ) . Conclusion The mechanism by which Tanshinonne ⅡA preconditioning reduces OGD∕R injury to N9 cells is related to inhibiting NF-κB signaling pathway.
3.Research progress on clinical application of ruxolitinib
Shiquan FENG ; Zhenmiao QIN ; Xue HU ; Deqiao DONG ; Haoyang PENG ; Changran GAN ; Chengcheng DUAN ; Yanan GAO
China Pharmacy 2024;35(13):1668-1672
Ruxolitinib, a small molecule inhibitor, selectively targets Janus kinase (JAK) by competitively binding to adenosine triphosphate on the catalytic site of the JAK1 and JAK2 domain, thereby inhibiting JAK activation and signal transducer and activator of transcription (STAT) phosphorylation and prevents the expressions of the JAK-STAT signaling pathway. Oral ruxolitinib has demonstrated promising efficacy for myelofibrosis and polycythemia vera. The topical Ruxolitinib cream, approved by the US FDA as the first non-segmental vitiligo home treatment drug, is set to be launched in domestic medical pioneer areas in August 2023 and is expected to bring about a breakthrough in the treatment of vitiligo. Clinical cases have also shown that Ruxolitinib cream has significant curative effects on atopic dermatitis, alopecia areata, and other conditions, indicating great application prospects.