Reference Intervals of Mitochondrial DNA Copy Number in Peripheral Blood for Chinese Minors and Adults / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mitochondrial DNA (mtDNA) content measured by different techniques cannot be compared between studies, and age- and tissue-related control values are hardly available. In the present study, we aimed to establish the normal reference range of mtDNA copy number in the Chinese population.</p><p><b>METHODS</b>Two healthy cohorts of 200 Chinese minors (0.1-18.0 years) and 200 adults (18.0-88.0 years) were recruited. Then, they were further categorized into eight age groups. The absolute mtDNA copy number per cell was measured by a quantitative real-time polymerase chain reaction. We subsequently used this range to evaluate mtDNA content in four patients (0.5-4.0 years) with molecularly proven mitochondrial depletion syndromes (MDSs) and 83 cases of mitochondrial disease patients harboring the m.3243A>G mutation.</p><p><b>RESULTS</b>The reference range of mtDNA copy number in peripheral blood was 175-602 copies/cell (mean: 325 copies/cell) in minors and 164-500 copies/cell (mean: 287 copies/cell) in adults. There was a decreasing trend in mtDNA copy number in blood with increasing age, especially in 0-2-year-old and >50-year-old donors. The mean mtDNA copy number level among the mitochondrial disease patients with m.3243A>G mutation was significantly higher than that of healthy controls. The mtDNA content of POLG, DGUOK, TK2, and SUCLA2 genes in blood samples from MDS patients was reduced to 25%, 38%, 32%, and 24%, respectively.</p><p><b>CONCLUSIONS</b>We primarily establish the reference intervals of mtDNA copy number, which might contribute to the clinical diagnosis and monitoring of mitochondrial disease.</p>

Fluid shear stress regulates epithelial-mesenchymal transition (EMT) in Hep2 cells / 医用生物力学

Objective To examine the effects of fluid shear stress (FSS) on epithelial-mesenchymal transition (EMT) in Hep2 cells. Methods Hep2 cells were exposed to 140 mPa FSS. The morphologic changes of Hep2 cells exposed to FSS at different durations were observed using inverted microscope. The migration ability of Hep2 cells after FSS loading was investigated using scratch wound assay. The distribution and expression of cytoskeleton protein F-actin were examined by confocal microscope. The expression of the EMT marker proteins were detected by Western blotting. Results Most of Hep2 cells changed their morphology from polygon to elongated spindle with well-organized F-actin under FSS. After removing FSS, Hep2 cells recovered their initial morphology with flat polygon. FSS regulated Hep2 cells to enhance their migration capacity in a time-dependent manner. FSS promoted the rearrangement of cytoskeletal protein F-actin，which enhanced the migration behavior of Hep2 cells. In addition, FSS induced a time regularity of expression of the EMT marker proteins in Hep2 cells. Conclusions FSS as an important physical factor can induce EMT in Hep2 cells.

Association between mthfr gene polymorphisms and toxicity of HDMTX chemotherapy in acute lymphocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate the association between mthfr gene polymorphisms and toxicity of HDMTX in acute lymphocytic leukemia patients. A total of 44 patients were selected, and DNA was extracted from their peripheral blood. PCR-RFLP was used to determine the genotypes of mthfr. The toxicity response of patients received HDMTX chemotherapy was observed. The results showed that the toxicity of HDMTX to carriers of the variant allele at codon 677 (CT or TT) increased, as compared with individuals with the common CC genotype (OR = 3.75, 95% CI 1 - 14, p = 0.04). In contrast, the toxicity of HDMTX to ALL patients with the variant allele at codon 1298 (AC or CC) decreased as compared with the common AA genotype carriers (OR = 0.12, 95% CI: 0.026 - 0.564, p = 0.007). As compared with carriers of the variant allele at coden 1298 (AC or CC), the toxicity of HDMTX to patients with TT genotype at 677 and AA genotype at 1298 increased (OR = 16.5, 95% CI: 0.026 - 0.564). It is concluded that mthfr gene polymorphisms associate with the toxicity of HDMTX chemotherapy in acute lymphocytic leukemia.

Preventive effects of emodin on cerebral ischemia injury and expression of the inflammatory factors in rats with cerebral ischemia / 中国中药杂志

<p><b>OBJECTIVE</b>To assess emodin antagonism to cerebral ischemia injury, and to discuss the mechanism of emodin inhibiting the inflammatory cascade reaction from the levels and expressions of cytokines.</p><p><b>METHOD</b>Rats were divided into sham-operated group, model group, Ligustrazine group and emodin groups (low, middle, high dosage). After focal cerebral ischemic model of cerebral middle artery occlusion was duplicated with nylon thread, we took the speciments after ischemia 6 hours, observed the changes of the evaluating score of neural symptoms, brain water ratio and cerebral infarction area, determined the levels of TNF-alpha, IL-beta and TGF-beta in rats brain tissue by radioimmunoassay, detected the expressions of TNF-alpha and VCAM-1 by immunohistochemistry, and measured VCAM-1-mRNA expression by in-situ hybridization.</p><p><b>RESULT</b>Compared with sham-operated group, the evaluating score of neural symptoms, brain water ratio and cerebral infarction area of rats in model group were higher (P < 0.01) , the levels of TNF-alpha and IL-1beta of rats brain tissue in model group increased, while the level of TGF-beta was lower, and the expressions of TNF-alpha and VCAM-1 increased (P < 0.01). The evaluating score of neural symptoms, brain water ratio and cerebral infarction area improved obviously in every emodin group, especially in emodin low dosage group. Levels of TNF-alpha, IL-1beta and the expressions of TNF-alpha and ICAM-1 in emodin low dosage group and Ligustrazine group were lower, while the level of TGF-beta was higher. Compared with Ligustrazine group, the changes aboved are more significant in emodin low dosage group (P < 0.01).</p><p><b>CONCLUSION</b>The increase of inflammatory cascade reaction mediated by various cytokines such as TNF, IL-1beta, ICAM-1 and the decrease of TGF protection are the important mechanism of cerebral ischemia injury. The mechanism of emodin antagonism to cerebral ischemia injury may be implemented by inhibiting inflammatory cascade reaction and increasing the brain protective factors, such as TGF.</p>

High vaginal uterosacral ligament suspension for treatment of uterine prolapse / 中华妇产科杂志

Objective To evaluate the physiological and anatomic basis,indications,surgical skills, prevention of ureter injury and clinic outcomes of using high uterosacral ligament suspension(HUS)for correction of advanced uterine prolapse by the vaginal route.Methods Fifty women with advanced uterine prolapse underwent transvaginal HUS after vaginal hysterectomy with reconstruction of pubocervical and rectovaginal fascia to correct their uterine prolapse between June 2003 and September 2007.The average age of the women was 60.1 years.The mean follow-up period was 24 months(range 4-51 months).The degree of pelvic organ prolapse preoperatively and anatomic outcomes postoperatively were assessed with pelvic organ prolapse quantification system(POP-Q).Results The remnants of the uterosacral ligaments were clearly identified and palpated posterior and medial to the ischial spines by traction with a 24 cm long Allis clamp and used for successful vaginal vault suspension and reconstruction in all 50 consecutive advanced uterine prolapse patients.The ureter injury was avoided by complete knowledge of the ureter's course from the cervix/apex toward its insertion in the sacral region and how far outside of the uterosacral ligament,by uteri palpation and by suturing purposefully placed"deep"dorsally and posteriorly toward the sacrum,as well as by cystoscopy examination of the spillage of urine from both ureters.Mean POP-Q point C improved from 1.5 to-7.5 cm with a median follow-up of 24 months.If the successful HUS was defined as point C≤stage I prolapse,both the objective and subjective cure rates were as high as 100% with a maximum follow-up of 51 months.None of the 50 patients had repeat operation for recurrence of prolapse.There was no major intra-or postoperative complications,such as ureter and other pelvic organ injury.Conclusion HUS with fascial reconstruction seems to be a safe,minimal traumatic,tolerable and highly successful procedure for vaginal repair of advanced uterine prolapse.Because of the use of native tissue as suspension site HUS is more physiologic and cost effective.