1.An experiment control study on the ovarian reserve function after cisplatin intraperitoneal or intravenous chemotherapy in rats model
Bozhen FAN ; Hong XIA ; Lei CHU ; Xiaowen TONG
Chinese Journal of Obstetrics and Gynecology 2017;52(4):249-253
Objective To compare the impact on the ovarian reserve function after cisplatin intraperitoneal or intravenous chemotherapy in rats model. Methods Thirty 8-weeks old female Sprague Dawley rats were randomly assigned to control group (group A, n=10), intraperitoneal chemotherapy group (group B, n=10) and intravenous chemotherapy group (group C, n=10). Cisplatin was diluted by normal saline (NS) into 4 mg/ml. On the first day of chemotherapy, 0.2 ml cisplatin dilution was injected into the abdomen of rats in group B, isodose cisplatin was injected into vein and 1.8 ml NS was injected into abdomen of rats in group C, 2.0 ml NS was injected into abdomen of rats in group A for control. Feed the three groups rats and test the anti-Mullerian hormone (AMH) in serum on day 0 (just before injection), day 10 and day 20 by ELISA, count the numble of follicle in bilateral ovaries on day 20. Results (1) The levels of serum AMH in the three groups before and after chemotherapy were compared: ① comparison between groups: On day 10 and day 20 , the AMH level in group B [(64.5 ± 2.9), (68.6 ± 3.4) ng/L] and group C [(76.1±4.9), (91.3±3.9) ng/L] was significantly lower than that in group A [(120.1±5.3), (121.7±4.6) ng/L;P<0.01], AMH level in group B was significantly also lower than that in group C (P=0.000). ② Comparison within groups:the AMH level on day 0 was significantly lower than that on day 10 and day 20 in group A (P<0.01), but there was no significant difference between day 10 and day 20 (P=0.427). The AMH level on day 0 was significantly higher than those on day 10 and day 20 in group B (P<0.01) and group C (P<0.01). There was no difference in AMH level between day 10 and day 20 (P=0.124) in group B, but the level was significant lower on day 10 than that on day 20 in group C (P=0.011). (2)Comparison of the number of follicles in ovaries of three groups 20 days after chemotherapy:the follicles number in group A(35±13)was greater than that in group B (16 ± 9,P=0.003) and similar with group C (31 ± 16,P=0.474) on day 20. The follicles number in group B was significantly less than that in group C (P=0.018). Conclusions In the present study, both intravenous and intraperitoneal chemotherapy have impacts on ovarian reserve function and the latter might be more serious. The level of AMH will rise again over time after chemotherapy and rats undergo intravenous chemotherapy would recover faster. The results suggest that the ovarian reserve function need more time to recover after intraperitoneal chemotherapy. For patients with ovarian cancer who want to preserve fertility function, intravenous chemotherapy might be more appropriate.
2.Deficiency in Na-K-2Cl co-transporter impaired hearing and balance in mice
Hanqi CHU ; Hao XIONG ; Fang HAN ; Zhengong WU ; Xiaowen HUANG ; Yonghua CUI
Chinese Journal of Pathophysiology 1999;0(09):-
AIM:We generated transgenic mice of NKCC1-/-(homozygous mutant),NKCC1+/-(heterozygous)and NKCC1+/+(wild-type)that have a targeted disruption in the NKCC1 gene to investigate the role of Na-K-2Cl(NKCC1)channel in auditory function of the inner ear.METHODS:Hearing threshold and endocochlear potential(EP)were measured in the NKCC1-/-,NKCC1+/-and NKCC1+/+ mice by auditory brainstem response(ABR)and EP recordings,respectively.The inner ears of the mice were removed and examined morphologically with the light microscope.RESULTS:The auditory function of NKCC1+/+ mice was normal,the mean value for ABR thresholds in response to click sound was [(23.13?3.78)dB,SPL],EP was(98?16)mV.The mean value for ABR thresholds to click sound was elevated in NKCC1+/-mice [(38.49?12.29)dB,SPL],relative to that significantly increased in NKCC1+/+ mice(P
3.Localization of NKCC1 in the cochlea and morphology of the cochlea in NKCC1-knockout mice.
Hanqi, CHU ; Hao, XIONG ; Xiaoqin, ZHOU ; Xiaowen, HUANG ; Liangqiang, ZHOU ; Yonghua, CUI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(3):374-7
The distribution of the Na-K-2Cl co-transporter (NKCC1) in the cochlear K+ cycling pathway in cochlea and cochlear histological changes in the NKCC1 knockout mice were investigated. By using immunohistochemistry and toluidine blue staining, the localization of NKCC1 in cochlea of the C57BL/6J mice and the cochlear histological changes in the NKCC1 knockout mice were observed. It was found that the NKCC1 was expressed mainly in the stria marginal cells and the fibrocytes in the inferior portion of the spiral ligament in the adult C57BL/6J mice. Subpopulation of the fibrocytes in the suprastrial region and the limbus was also moderately immunoreactive. While in the cochlea of the NKCC1 knockout mice, Reissner's membrane was collapsed and scala media disappeared, accompanied with the loss of inner hair cells, outer hair cells and the support cells. The tunnel of Corti was often absent. All the findings suggested the localization of NKCC1 in the cochlea was closely correlated with cochlear K+ cycling. Loss of NKCC1 led to the destruction of the cochlear structures, and subsequently influenced the physiological function of cochlea.
4.Study on p21 gene knock out in G401 cell line by using CRISPR/Cas9 system
Xiujuan ZHAO ; Wanbiao CHEN ; Peitao ZHANG ; Na ZHANG ; Xiaowen CHU ; Xiangyang BAI ; Bing YANG ; Xudong WU ; Xi WANG
Tianjin Medical Journal 2016;44(10):1190-1194
Objective To knock out p21 gene in human malignant rhab doid tumor(MRT)cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene,which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group,the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors .
5.Correlation of PDCD5 and apoptosis in hair cells and spiral ganglion neurons of different age of C57BL/6J mice.
Yan, WANG ; Hanqi, CHU ; Liangqiang, ZHOU ; Heyun, GAO ; Hao, XIONG ; Qingguo, CHEN ; Jin, CHEN ; Xiaowen, HUANG ; Yonghua, CUI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(1):113-8
This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice. Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3, 6, 9 or 12 months). PDCD5 expression was detected by using immunohistochemistry, real-time PCR and Western blot. Morphological change of the cochleae was also evaluated by using immunoassay. The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice, as well as gradually increased apoptosis of cochlear hair cells and SGNs. In addition, we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing. It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs, and thereby plays a role in the pathogenesis of presbycusis. Thus, PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.
6.Comparative analysis chest CT 3D reconstruction and esophagus barium swallow in esophagus foreign body.
Kai XU ; Hanqi CHU ; Xiaowen HUANG ; Yonghua CUI
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;24(12):549-550
OBJECTIVE:
To compare the diagnosis and therapeutic between chest computed tomography three-dimensional reconstruction and esophagus barium swallow in esophagus foreign body.
METHOD:
retrospective analyze one hundred and thirty six patients who suffered from esophagus foreign body in our hospital, 97 cases using esophagus barium swallow, 17 cases using chest computed tomography three-dimensional reconstruction, 15 cases using both.
RESULT:
The patients who showed positive of esophagus foreign body in esophagus barium swallow or chest computed tomography three-dimensional reconstruction, 91.8% (89/97) cases or 88.2% (15/17) cases found esophagus foreign bodies finally. All cases successfully took out the esophagus foreign bodies only through one operation which used chest computed tomography three-dimensional reconstruction as primary examination, while only 91.0% for those used esophagus barium swallow as primary examination.
CONCLUSION
Both chest computed tomography three-dimensional reconstruction and esophagus barium swallow showed high diagnostic efficiency on esophagus foreign body. Chest computed tomography three-dimensional reconstruction had advantages in patients with one of following conditions: (1) esophagus foreign body located in the middle of the esophagus, especially complicated with esophagus perforation; (2) with fever, high white blood count, presence of abscess surrounding the esophagus was suspected; (3) with dyspnea; (4) with a history of esophagus foreign body longer than 5 days; (5) younger than 6 years old.
Adolescent
;
Adult
;
Aged
;
Aged, 80 and over
;
Barium Sulfate
;
Child
;
Child, Preschool
;
Esophagus
;
diagnostic imaging
;
Female
;
Foreign Bodies
;
diagnostic imaging
;
Humans
;
Image Processing, Computer-Assisted
;
Imaging, Three-Dimensional
;
Infant
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Male
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Middle Aged
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Retrospective Studies
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Tomography, X-Ray Computed
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methods
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Young Adult
7.Correlation of PDCD5 and Apoptosis in Hair Cells and Spiral Ganglion Neurons of Different Age of C57BL/6J Mice
WANG YN ; CHU HANQI ; ZHOU LIANGQIANG ; GAO HEYUN ; XIONG HAO ; CHEN QINGGUO ; CHEN JIN ; HUANG XIAOWEN ; CUI YONGHUA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(1):113-118
This study examined the expression pattern of programmed cell death 5 (PDCD5) in cochlear hair cells and spiral ganglion neurons (SGNs) and its association with age-related hearing loss in mice.Sixty C57BL/6J (C57) mice at different ages were divided into four groups (3,6,9 or 12 months).PDCD5 expression was detected by using immunohistochemistry,real-time PCR and Western blot.Morphological change of the cochleae was also evaluated by using immunoassay.The results showed that the expression of PDCD5 had a gradual increase with ageing in both protein and RNA levels in C57 mice,as well as gradually increased apoptosis of cochlear hair cells and SGNs.In addition,we also found that caspase-3 activity was enhanced and its expression was enhanced with ageing.It is implied that overexpression of PDCD5 causes the increase in caspase-3 activity and the subsequent increase of apoptosis in cochlear hair cells and SGNs,and thereby plays a role in the pathogenesis of presbycusis.Thus,PDCD5 may be a new target site for the treatment and prevention of age-related hearing loss.
8.Localization of NKCC1 in the Cochlea and Morphology of the Cochlea in NKCC1-Knockout Mice
Hanqi CHU ; Hao XIONG ; Xiaoqin ZHOU ; Xiaowen HUANG ; Liangqiang ZHOU ; Yonghua CUI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(3):374-377
The distribution of the Na-K-2Cl co-transporter (NKCC1) in the cochlear K+ cycling pathway in cochlea and cochlear histological changes in the NKCC1 knockout mice were investigated. By using immunohistochemistry and toluidine blue staining, the localization of NKCC1 in cochlea of the C57BL/6J mice and the cochlear histological changes in the NKCC1 knockout mice were observed. It was found that the NKCC1 was expressed mainly in the stria marginal cells and the fibrocytes in the inferior portion of the spiral ligament in the adult C57BL/6J mice. Subpopulation of the fibrocytes in the suprastrial region and the limbus was also moderately immunoreactive. While in the cochlea of the NKCC1 knockout mice, Reissner's membrane was collapsed and scala media disappeared, accompanied with the loss of inner hair cells, outer hair cells and the support cells.The tunnel of Corti was often absent. All the findings suggested the localization of NKCC1 in the cochlea was closely correlated with cochlear K+ cycling. Loss of NKCC1 led to the destruction of the cochlear structures, and subsequently influenced the physiological function of cochlea.
9.Effects of cord blood element levels on neurodevelopment of preterm and full-term children: A cohort study
Zhaokun WANG ; Wenlou ZHANG ; Xiaowen ZENG ; Chu CHU ; Qingqing LI ; Xinxin CUI ; Qizhen WU ; Guanghui DONG ; Jinbo HUANG ; Minli KONG ; Furong DENG
Journal of Environmental and Occupational Medicine 2022;39(7):723-729
Background Essential and non-essential elements have an important impact on the development of the central nervous system during fetal development. Due to their less developed brain, preterm infants are more sensitive to element exposure, and are high-risk groups of neurodevelopmental abnormalities. However, it is not clear whether the effects of element exposure in utero on postpartum neurodevelopment are different between full-term infants and preterm infants. Objective To evaluate the effects of element exposure levels during pregnancy on neurodevelopment of children aged 6-24 months (of corrected age), and compare the effects between preterm and full-term children. Methods A prospective study design was adopted and this study was conducted based on the Maoming Birth Cohort Study (MBCS) in Maoming City, Guangdong Province. Twenty elements in cord blood of 197 preterm infants and 297 full-term infants were measured, including 11 essential trace elements [vanadium (V), chromium (Cr), manganese (Mn), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), selenium (Se), strontium (Sr), tin (Sn), and iron (Fe)], and 9 non-essential trace elements [aluminum (Al), arsenic (As), thallium (Tl), lead (Pb), uranium (U), cerium (Ce), antimony (Sb), cadmium (Cd), and yttrium (Y)]. The neurodevelopment of the children at 6, 12, and 24 months were evaluated by the Ages and Stages Questionnaires-the Third Edition (ASQ-3). A generalized estimating equation (GEE) model was adopted to evaluate the associations between elements and neurodevelopment in full-term and preterm children separately. Results The positive rates of 10 elements (Mn, Cu, Zn, Se, Sr, Fe, Sb, Tl, Pb, and As) in cord blood were greater than 80%. Among the preterm birth children, the results of GEE analysis showed that after adjusting for the covariates, for each increase of interquartile range (IQR) in ln-transformed concentration, As was associated with problems/delay in the communication and problem-solving sub-scales, with the adjusted odds ratios (OR) and 95% confidence intervals (CI) of 1.36 (1.03-1.80) and 1.55 (1.10-2.20), respectively; the adjusted OR (95%CI) of problems/delay in the fine motor and problem-solving sub-scales were 1.44 (1.00-2.07) and 1.76 (1.09-2.84) for Sb, respectively; the adjusted OR (95%CI) of problems/delay in the communication sub-scale was 1.37 (1.09-1.74) for Se. No statistically significant associations between umbilical cord blood element concentrations and neurodevelopment indicators were observed among full-term children. The results of stratified analysis by sex showed that the associations between umbilical cord blood element concentrations and neurodevelopment problems/delay were only significant among female preterm children. Conclusion Exposures to As, Se, and Sb during pregnancy may increase the risk of neurodevelopment problems/delay in preterm children aged 6-24 months, and female seem to be more vulnerable.