2.Changes of P-selectin and E-selectin in children with Kawasaki disease.
Ye QIU ; Jie WU ; Xiao-yi FANG ; Zhen LIN ; Bei-yan WU ; Ruo-yin CAI ; Xiao-yan XU ; Hong ZHENG
Chinese Journal of Pediatrics 2004;42(9):688-692
OBJECTIVEKawasaki disease (KD) is a kind of febrile disorder without definite etiology. The pathologic change of KD is characterized by nonspecific vasculitis, which mainly involves the coronary artery. Some patients may have coronary angioma formation, and some of them will result in the coronary narrowing or embolism. Notwithstanding that KD has been one of the most common causes for acquired heart diseases in childhood in addition to the rheumatic fever, the pathogenesis of the vascular damage remains unknown. This study was conducted to explore the pathophysiological role of cell adhesion molecules (P-selectin and E-selectin) on the endothelial lesions in KD, and to look for the evidence of direct relationship between the plasma levels of soluble cell adhesion molecules (P- and E-selectin) and the incidence of the coronary artery lesion (CAL).
METHODSSoluble P-selectin (PS), E-selectin (ES), thromboxane-B(2)(TXB(2)), 6-keto-PGF(1)alpha (6-KPGF(1)alpha) were measured in 36 patients with KD, 20 patients with febrile disease and 30 healthy children by using double antibody sandwich enzyme linked immunosorbent assay (ELISA) and radioimmunoassay. Patients with KD were separated into acute phase group, subacute phase group, recovery phase group, coronary artery lesion group (CAL), non-coronary artery lesion group (NCAL), intravenous immunoglobulin (IVIG) effective group (body temperature back to normal after 48 hours of using IVIG), and IVIG ineffective group.
RESULTSPlasma PS and ES levels in the acute phase group [(211 +/- 28 and 186 +/- 14) ng/ml], subacute phase group [(238 +/- 27 and 151 +/- 13) ng/ml] and recovery phase group [(198 +/- 21 and 1008 +/- 9) ng/ml] were significantly higher than those in the healthy group [(102 +/- 36 and 72 +/- 10) ng/ml, P < 0.01]. The plasma PS levels remained higher after the treatment, but in IVIG effective group, the PS and ES levels declined significantly (P < 0.01) compared with those in acute phase group. Plasma PS and ES levels of CAL group [(281 +/- 78 and 210 +/- 52) ng/ml] were significantly higher than those of NCAL group [(217 +/- 15 and 108 +/- 10) ng/ml, P < 0.01]. In contrast to 1 week after the treatment, the PS and ES in IVIG effective group at the time point of 2 weeks after the treatment decreased more significantly (P < 0.01). While the PS and ES in IVIG ineffective group remained higher at the time point of 2 weeks after the treatment, which showed no significant difference compared with those 1 week after the treatment (P > 0.05). One week after the treatment, the PS levels of IVIG effective and ineffective groups did not descend, and there was no significant difference in PS between these two groups at this time point. Two weeks after the treatment, the PS and ES in IVIG ineffective group remained higher than those in IVIG effective group, and there was a significant difference between them. The peak level of PS appeared in the subacute phase. TXB(2) levels of KD in acute phase group increased markedly, which were significantly higher than those of healthy group [(345 +/- 127 and 190 +/- 69) ng/L, P < 0.01]. There was no significant difference between subacute phase group and healthy group. No significant difference was found between CAL group and NCAL group (P > 0.05). The levels of TXB(2) declined quickly after the treatment. The 6-KPGF(1)alpha level in KD of acute phase group, subacute phase group and recovery phase group [(7.1 +/- 2.8, 10.8 +/- 3.7 and 11.3 +/- 4.0) ng/L, respectively] was significantly lower than that of healthy group [(17.7 +/- 5.8) ng/L, P < 0.01], and the levels did not recover to normal even 2 weeks after the treatment. There was no significant difference 6-KPGF(1)alpha levels between CAL group and NCAL group (P > 0.05). In the febrile group, PS and ES levels showed no significant differences compared with healthy children (P > 0.05). ES level of KD patients was significantly correlated with CRP levels (r = 0.79 P < 0.01). In febrile group, there was no significant correlation between ES and CRP. There was a significant correlation between PS and PLT levels in KD patients (r = 0.75 P < 0.01), and no significant correlation between PS and PLT levels in febrile patients.
CONCLUSIONThe increase of plasma PS and ES levels in KD acute phase and subacute phase might play an important role in the pathophysiology of the endothelial damage. E- and P-selectin may potentially be a predictor of CAL in patients with KD.
Child ; Coronary Artery Disease ; physiopathology ; Coronary Vessels ; physiopathology ; E-Selectin ; blood ; Endothelium, Vascular ; physiopathology ; Humans ; Mucocutaneous Lymph Node Syndrome ; blood ; physiopathology ; P-Selectin ; blood
3.Molecular genetic analysis of congenital lipoid adrenal hyperplasia.
Wen-Juan QIU ; Jun YE ; Bei HAN ; Lian-Shu HAN ; Xue-Fan GU
Chinese Journal of Pediatrics 2004;42(8):585-588
OBJECTIVECongenital lipoid adrenal hyperplasia (CLAH) is an autosomal recessive inherited disorder, characterized by deficiency of adrenal and gonadal steroid hormones. Recent studies have shown that mutations in the gene for steroidogenic acute regulatory protein (StAR) cause this most severe genetic disorder in steroid hormone biosynthesis. StAR is a mitochondrial protein promotes cholesterol transfer from outer mitochondrial membrane to the inner mitochondrial membrane, where the cholesterol serves as a substrate for P450scc and initiates steroidogenesis. So far, more than 30 different mutations in the StAR gene have been found in the patients with CLAH from various ethnic groups. None of CLAH patients in the Chinese population has been previously reported. In the present study we analyzed the StAR gene in a Chinese patient with CLAH.
METHODSThe patient who was a 19-yr-old phenotypic female, has a 46, XY karyotype. Endocrinological evaluation was performed. Genomic DNA samples were abstracted from the bloods of the patient and his parents. Polymerase chain reaction (PCR), direct DNA sequencing, family analysis and restriction enzyme digestion analysis were used to detect and confirm the mutations of StAR gene.
RESULTSEndocrine evaluation of the patient showed extremely elevated basal concentrations of serum ACTH and gonadotropin and minimal concentration of gonadal steroids. An ACTH stimulation test indicated basal serum dehydroepiandrosterone and 17-hydroxyprogesterone were lower than normal detectable range and had no obvious increase after the ACTH stimulation. Automatic sequencing of 7 exons of the StAR gene with the polymerase chain reaction products of the genomic DNA revealed compound heterozygous for a novel nonsense mutation Q77X in exon 3 and the frameshift mutation 838delA in exon 6. The father carried Q77X mutation and the mother carried 838delA mutation. The restriction enzyme site of the Q77X mutation was examined by endonucleotidase BfaI. Furthermore, this mutation was not found in a series of 20 alleles of normal individuals.
CONCLUSIONQ77X is the novel mutation found in the patient with CLAH. Q77X and 838delA compound mutations could inactivate the StAR function and give rise to clinically manifest CLAH. This case is the first Chinese patient with CLAH identified by molecular genetic analysis. DNA-based analysis of StAR gene will be helpful for the diagnosis of CLAH.
Adrenal Hyperplasia, Congenital ; complications ; genetics ; Adrenal Insufficiency ; etiology ; Female ; Genotype ; Gonadal Steroid Hormones ; deficiency ; Humans ; Mutation ; Phenotype ; Phosphoproteins ; genetics ; Young Adult
4. Clinical evaluation of cervicectomy in the treatment of cervical intraepithelial neoplasia with cervicovaginal shortening
Yue QI ; Qiu-lin YE ; Miao LIU ; Pei-yao WANG ; Hui-min WANG ; Juan-juan LIU ; Lian-cheng ZHU ; Bing-ying LIU ; Dan-ye ZHANG ; Bei LIN
Chinese Journal of Practical Gynecology and Obstetrics 2019;35(10):1130-1136
OBJECTIVE: To investigate the clinical efficacy of cervicectomy for cervical intraepithelial neoplasia(CIN)in patients with cervicovaginal shortening.METHODS: Retrospectively analyze the clinical data of the 120 cases of CIN treated in Shengjing Hospital of China Medical University from April 2014 to November 2018.Cervicectomy was performed because of cervicovaginal shortening caused by menopausal or peri-menopausal cervical atrophy or cervical surgery.The clinical treatment,efficacy and prognosis of the patients were reviewed.RESULTS: The mean age of the 120 patients was 55.2 years(range:35-77 years).The indications of operation included:persistent abnormal cervical cytology test(7),CIN2(42),CIN3(70),squamous carcinoma of the cervix(1);peri-menopausal and menopausal patients with obvious cervicovaginal atrophy(100),premenopausal patients with natural short cervix(2),and obvious cervicovaginal shortening caused by cervical surgery(18).The mean operating time was 23.2 min(range 10-30 min),the mean bleeding volume was 7.8 mL(range:5-20 mL),and the mean height of cervix resected was 2.59 cm(range:2-3 cm).No secondary injury,bleeding or other postoperative complications occurred during surgery;cervical postoperative wounds healed well;only one case developed cervical adhesion after surgery.The postoperative histologic diagnosis were compared with the preoperative histologic diagnosis,in which 45 degraded(37.50%),42 consistent(35.00%),and 33 upgraded(27.50%).HPV conversion rate 3 months after cervicectomy was 80.81%(80/99),and total HPV conversion rate was 88.89%(88/99).A total of 29 patients underwent secondary surgery,23 underwent total hysterectomy,and 6 underwent extensive hysterectomy and pelvic lymphadenectomy.All patients were followed up,once every 3 to 6 months,and median follow-up time was 29.5 months(range 4-59 months).All patients recovered well after surgery;only 2 cases showed positive margins,and only 2 cases of residual disease and 1 case of recurrence were found during follow-up.CONCLUSION: For patients with cervical intraepithelial neoplasia of cervicovaginal shortening,cervicectomy is a safe,effective and relatively microinvasive treatment.
5.Casticin Attenuates Stemness in Cervical Cancer Stem-Like Cells by Regulating Activity and Expression of DNMT1.
Xue-Li WANG ; Xiao-Zheng CAO ; Dao-Yuan WANG ; Ye-Bei QIU ; Kai-Yu DENG ; Jian-Guo CAO ; Shao-Qiang LIN ; Yong XU ; Kai-Qun REN
Chinese journal of integrative medicine 2023;29(3):224-232
OBJECTIVE:
To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism.
METHODS:
Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L).
RESULTS:
DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells.
CONCLUSION
CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.
Female
;
Humans
;
Cell Line, Tumor
;
HeLa Cells
;
Neoplastic Stem Cells/metabolism*
;
RNA, Small Interfering/metabolism*
;
Uterine Cervical Neoplasms/metabolism*