1.Aurora-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells
Yi ZHANG ; Dongqin CHEN ; Kai ZHANG ; Rui WANG ; Longbang CHEN
Journal of Medical Postgraduates 2015;(6):579-583
Objective Human lung adenocarcinoma SPC-A1/DTX cells have a higher radioresistance than SPC -A1cells. This study was to investigate the role of Aurora-An/uclear factor κB ( NF-κB) in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells and its possible molecular mechanisms . Metho ds We collected human lung adenocarcinoma SPC-A1 and SPC A1/DTX cells and divided them into four groups:sh-Aurora-A ( Aurora-A plasmid interference ) , sh-NC, NF-κB inhibition ( SPC-A1/DTX +NF-κB inhibitor ) , and DMSO control .We measured the in vitro radio-sensitivity of the cells by MTT assay , determined their proliferation ability by cloning assay , and detected the mRNA and protein expressions of the target genes by real -time quantitative RT-PCR and Western blot , respectively . Results The 50% effective doses ( ED50 ) of the SPC-A1 and SPC-A1/DTX cells on radiotherapy were (6.5 ±0.3) and (12.8 ±0.6) Gy, respectively, with statisti-cally significant difference between the two groups ( P <0.01 ) .In the radiation doses of 0, 2, 4, and 6 Gy, the numbers of the cloned SPC-A1 cells were 345 ±20 , 252 ±22 , 170 ±15 , and 81 ±10 , sig-nificantly lower than those of the cloned SPC -A1/DTX cells (402 ±21, 370 ±18, 301 ±16, and 252 ±15) (P<0.05).The protein and mRNA expressions of Aurora-A were remarkably higher in the SPC-A1/DTX than in the SPC-A1 cells (1.00 ±0.08 and 1.00 ±0. 06 vs 0.49 ±0.03 and 0.22 ±0.02, P<0.05).MTT assay showed a higher ED50 in the sh-NC than in the sh-Aurora-A cells ([11. 8 ±0.5] vs [7.1 ±0.3] Gy, P<0.01) as well as in the control than in the NF-κB inhibition group ([11.7 ±0.5] vs [6.1 ±0.3] Gy, P<0.01).Inhibition of Aurora-A increased the expression of IκBa by 2.18 ±0.32 times (P<0.01) and that of NF-κB by 0.24 ±0.03 times (P<0.01).The expressions of IκBa (1.00 ±0.05) and NF-κB (1.00 ±0.04) were significantly lower in the parent strains of SPC-A1 than 0.65 ±0.04 and 2.18 ±0.15 in the drug-resistant strains of SPC-A1/DTX (P<0.01). Conclusi on Auro-ra-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells.
2.Research progress of small molecule fluorescent probes for ferrous ion and heme
Chen CHEN ; Yi-xin CHEN ; Chong-jing ZHANG
Acta Pharmaceutica Sinica 2023;58(8):2250-2259
Small molecule fluorescent probes have gained widespread attention for their advantages of high selectivity, sensitivity, and easy to operate, and have played a critical role in the detection of various species. They have also demonstrated great potential in the field of biomedical research. Iron, as the most abundant transition metal in the human body, plays a vital role in many physiological functions. Due to the influence of the reductive microenvironment of cell, ferrous ion (Fe2+) is the main component of labile iron in living cells. Heme, consisting of Fe2+ and protoporphyrin IX, is one of the main signaling molecules that wrap biological iron in the human body, and also participates in many physiological and pathological processes. Therefore, the development of small molecule fluorescent probes for detecting Fe2+ and heme as effective monitoring tools will help to further understand their pathological and physiological functions, with potential applications in other fields. This review summarizes the research progress of small molecule fluorescent probes for Fe2+ and heme detection in recent years, and provides insights into future directions for their development.
3.Effects of perindopril on bone metabolism in a rat model of retinoic acid-induced osteoporosis
Yi ZHONG ; Qing XUE ; Yi ZHOU ; Lianghua CHEN ; Lichao ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(18):2589-2595
BACKGROUND: Renin-angiotensin-aldosterone system existed in bone tissue. Recent studies on antihypertensive drugs found that angiotensin converting enzyme inhibitor type antihypertensive drug was possibly effective for osteoporosis. Perindopril is one of the commonly used antihypertensive drugs. Whether perindopril affected bone metabolism or could be used in anti-osteoporosis has not been reported.
OBJECTIVE: To observe effects of perindopril on bone metabolism in a rat model of osteoporosis induced by retinoic acid.
METHODS: Fifty Sprague-Dawley rats were randomly divided into five groups, with ten in each group. In the model group and each perindopril groups, rats were intragastricaly administered retinoic acid solution 80 mg/kg per day. After successful model establishment, rats in different perindopril groups were intragastrical y administered perindopril 2, 4 and 8 mg/kg per day, once a day, for 42 consecutive days. In the normal control and model groups, rats were given an equal volume of distil ed water. Serum calcium, phosphorus, alkaline phosphatase, bone mass and bone mineral density were detected in each group. Expression of bone specific alkaline phosphatase and tartrate-resistant acid phosphatase mRNA in bone tissue was determined.
RESULTS AND CONCLUSION: Compared with the model group, after treatment with perindopril, serum calcium and phosphorus levels were increased, alkaline phosphatase activities were significantly decreased, bone mass and bone mineral density were obviously increased in rats with retinoic acid-induced osteoporosis. Expression of bone specific alkaline phosphatase mRNA was higher in the perindopril 8 mg/kg group than in the perindopril 2 and 4 mg/kg groups and model group. Tartrate-resistant acid phosphatase mRNA expression was higher in the perindopril 8 mg/kg group than in the model group. These results indicated that perindopril could improve partial bone metabolic biochemical markers in osteoporosis rats, promoted bone formation by up-regulating bone specific alkaline phosphatase mRNA expression, and had a certain preventive effect on retinoic acid-induced osteoporosis.
4.Research on the diagnostic values of the detection of urine exfoliated cells by FISH and cytologic technology in bladder urothelial tumor
Wei CHEN ; Huixia YI ; Rui ZHANG
International Journal of Laboratory Medicine 2014;(22):3094-3096
Objective To compare the diagnostic values of the detection of urine exfoliated cells by FISH and cytology technolo‐gy in bladder urothelial tumor .Methods The combination probes of CSP3/CSP7 and GLPp16/CSP17 were both used in the FISH detection of urine exfoliated cells from suspected patients with bladder urothelial tumor .The urine exfoliated cells were detected by cytology technology at the same time .The sensitivity and the specificity of the two methods were compared .Results The sensitivi‐ty and specificity of FISH for bladder urothelial tumor screening were 92 .5% and 85 .0% respectively ,and those of cytology tech‐nology were 27 .5% and 90 .0% respectively .The sensitivity of FISH was significantly higher than that of cytology technology (P<0 .05) ,however ,the specificity differences between FISH and cytology technology were not statistically significant (P>0 .05) .Conclusion FISH is expected to become a new method for the screening of bladder urothelial tumor .
5.Expressions of Survivin, MMP-2 and TIMP-2 in cervical carcinoma and their relationship with the invasion and metastasis of the cancerous tissues
Yi LUO ; Shuangyun CHEN ; Changju ZHANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(02):-
Objective To investigate the expression of survivin, matrix metalloproteinase (MMP-2) and tissue inhibitor 2 of matrix metalloproteinase (TIMP-2) in cervical carcinoma and their relationship with invasion and node metastasis of the cervical cancerous tissues. Methods The expressions of survivin, MMP-2 and TIMP-2 were examined by immunohistochemical S-P method and colour pathological image computer analysis system in 10 cases of normal cervical epithelia, 10 cases of cervical carcinoma in situ, 40 cases of invasive squamous cell cervical carcinoma and 11 cases of invasive cervical adenocarcinoma. The relationship between those indexes and the factors related to clinical pathology of cervical carcinoma were analyzed statistically. Results It was found that the positive level of survivin and MMP-2 expression increased in the order of normal cervical epithelium, cervical carcinoma in situ and invasive carcinoma of cervix (P0.05). The positive expressions of survivin and MMP-2 in patients under 35 years old or with pelvic lymph node metastasis, intravascular involvement and stroma involvement were significantly higher than that in the cases without them, while TIMP-2 expression was opposite to that of MMP-2 (P
6.Effects of different concentrations of ox-LDL on the proliferation of rat theca cells and the expression of steroidogenesis related genes LXR-α and StAR
Ying CHEN ; Yi ZHANG ; Cong LI
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(3):318-324
Objective · To investigate the effects of ox-LDL on the proliferation of rat theca cells and expression of LXR-α and StAR, two genes associated with androgen biosynthesis. Methods · The expression of LXR-α in the ovarian tissue of rats was determined by immunohistochemistry. Primary theca cells were isolated and collected from rat ovary and cultured in vitro. Furthermore, the theca cells were treated with 25, 50, 100, 150, 200, 300 and 400 mg/L ox-LDL, respectively. The variations in LXR-α mRNA were identified using real-time PCR. MTT assay was performed to detect cell viability. The expression of LXR-α and StAR was measured by Western blotting analysis. Results · The effect of ox-LDL on the proliferation of rat theca cells and the levels of LXR-α and StAR in theca cells was in a concentration-dependent manner. Following exposure to various concentration of ox-LDL for 24 h, the proliferation of theca cells was induced by low concentration of ox-LDL (25-150 mg/L), and 100 mg/L ox-LDL showed the most significant inducing effect. Moreover, the cell survival rate was diminished considerably following with ox-LDL concentration increasing, especially lowered by 400 mg/L ox-LDL. The mRNA level of LXR-α was increased with low concentration of ox-LDL (25-150 mg/L) and the impact of ox-LDL on the induced expression of LXR-α mRNA was considerably distinct at the concentration of 150 mg/L. On the other hand, the expression of LXR-α mRNA was reduced with high concentration of ox-LDL, and the impact of 400 mg/L ox-LDLwas substantially distinct. The protein expression levels of LXR-α and StAR were increased with 150 mg/L ox-LDL, but StAR protein level in 150 mg/L ox-LDL group revealed no significant difference when compared with control group. The expression of LXR-α and StAR protein was significantly inhibited with 400 mg/L ox-LDL in the rat theca cells. Conclusion · Low concentrations of ox-LDL can induce the proliferation of theca cells, and promote the expression of StAR and LXR-α. Whereas, high concentrations of ox-LDL can reduce the cell viability and inhibit the expression of StAR and LXR-α.
7.Determination of Content of Ceftizoxime by RP-HPLC
Aichun YI ; Shunzhi ZHANG ; Difang CHEN
China Pharmacy 2001;0(11):-
OBJECTIVE:To determine the content of ceftizoxime for injection METHODS:A RP-HPLC method was established,the polarls C18 was used as analysis column The mobile phase consisted of 0 23% sodium hydrogen phosphate and 0 14% trisodium buffer-acetonitrile(85∶15),the flow rate was 1 0ml/min,the detection wavelength was 254nm RESULTS:The linear range of ceftizoxime was 4 1 462~37 3 156?g/ml,r=0 9 999 The relative standard deviation was 0 6% CONCL_USION:The method is rapid,accurate and suitable for determination of content of ceftizoxime for injection
9.Extraction and Determination of Polysaccharide from Discarded Fi brous Roots of Radix Panacis Quingueforlii
Renquan ZHANG ; Jieping L ; Yi CHEN
Traditional Chinese Drug Research & Clinical Pharmacology 2001;12(2):109-110
Objective:To establish a feasible method for extracti ng polysaccharide from the discarded fibrous roots of Radix Panacis Quingueforli i afterpanaquilon had been extracted, and determine the polysaccharide content. Methods:Enzymolysis technique and alcohol was applied for decolorization, and ph enol-sulfuric-acid method was used to determine the active polysaccharide conte nt.The content of trace element and heavy mental was measured by element-analyze r and atomic fluorescence photometer respectively. Results: The yield of polysac charide from the fibrous roots was close (about 11.7%) to the main root.And the content of heavy metal can match the national standard.Conclusion:I t is valuable to extract the polysaccharide from the discarded fibrous root of R adix Panacis Quingueforlii.
10.Effect of preoperative splenectomy on liver transplantation in treatment of portal hypertension
Jingyu CHEN ; Yi JIANG ; Xiaojin ZHANG
Chinese Journal of Hepatobiliary Surgery 2014;20(8):572-576
Objective To study the influence of preoperative splenectomy on liver transplantation for portal hypertension and the management of complications.Methods From January 2008 to January 2013,a total of 136 patients underwent liver transplantation for portal hypertension (PHT) in Hepatobiliary Surgery Center,Fuzhou General Hospital of PLA Nanjing Military Region.Pre-liver transplantation splenectomy was carried out in 19 patients.We randomly selected according to the operation time 50 cases of liver transplantation as the control group.The operation time,bleeding,blood transfusion,infection,the incidence of acute rejection,biliary complication rate,survival rate,postoperative platelet recovery and arteriovenous thrombosis were analyzed.Results The preoperative group has longer surgical time,intraoperative blood loss,blood transfusion volume increased obviously,infection rate increased significantly compared with the control group.The accumulate survival rate of preoperative splenectomy group was significantly lower than that of the control group.The platelet count of the preoperative splenectomy group was significantly higher than that of the control group.The platelet count of both groups experienced the process of first rise after falling.Although the platelet count of the preoperative splenectomy group was higher than that of the control group,but there was no statistically significant difference.Conclusions Liver transplantation in PHT patients with previous splenectomy may lead to some unfavorable consequences including increases of technical diffculty of surgery,operative duration,intraoperative bleeding,postoperative infection and other serious complications,and surgical mortality.Therefore,splenectomy should be performed cautiously for the patients who may receive liver transplantation in the future.