1.Preliminary study on predicting early embryonic developmental potential by metabolomics analysis of day 3 embryo culture medium
Xuehua CHEN ; Jinhui SHU ; Kejian SUN ; Hong ZHOU ; Xiaojing FAN ; Caizhu WANG
Chinese Journal of Reproduction and Contraception 2025;45(4):380-389
Objective:To explore the differences of metabolomic profiles in day 3 (D3) culture medium of embryos with varying developmental potentials, in order to provide a theoretical foundation for the establishment of embryo selection technology platform using metabolomics.Methods:Eight patients who received in vitro fertilization and embryo transfer (IVF-ET) treatment at Reproductive Medicine Center of Guangxi Zhuang Autonomous Region Maternal and Child Health Hospital between November 13 and December 5, 2023 were selected as the study subjects. The D3 culture medium from patient embryos was collected and divided into high-quality blastocysts ( n=42), non-high-quality blastocysts ( n=33), and embryos that failed to form blastocysts (non-formation group, n=43) according to the formation of day 5 blastocysts. High-performance liquid chromatography-mass spectrometry was employed to perform non-targeted metabolomic analysis in the D3 culture medium from three distinct groups. Results:1) The metabolites in D3 culture medium of embryos with varying developmental potentials exhibit significant differences. Specifically, 79 differential metabolites were identified between the blastocyst formation group and the non-blastocyst formation group (all P<0.05); additionally, 73 differential metabolites were found between the high-quality blastocyst group and the non-high-quality blastocyst group (all P<0.05). 2) The area under the receiver operating characteristic curve of significantly differential metabolites for predicting potential of D3 embryo blastocyst formation and high-quality blastocyst formation were both greater than 0.9, demonstrating excellent predictive performance. 3) KEGG pathway enrichment analysis revealed that differential metabolites associated with blastocyst formation potential were primarily enriched in pathways including D-amino acid metabolism, glycine-serine-threonine metabolism, arginine biosynthesis, and histidine metabolism ( P<0.05). For high-quality blastocyst formation, the differential metabolites were predominantly enriched in pathways related to tryptophan metabolism, D-amino acid metabolism, serotonergic synapses, and protein digestion and absorption ( P<0.05). Conclusion:Embryos with different developmental potentials have significantly different metabolic profiles, and it is feasible to predict the developmental potential of D3 embryos by metabolomics analysis.
2.Preliminary study on predicting early embryonic developmental potential by metabolomics analysis of day 3 embryo culture medium
Xuehua CHEN ; Jinhui SHU ; Kejian SUN ; Hong ZHOU ; Xiaojing FAN ; Caizhu WANG
Chinese Journal of Reproduction and Contraception 2025;45(4):380-389
Objective:To explore the differences of metabolomic profiles in day 3 (D3) culture medium of embryos with varying developmental potentials, in order to provide a theoretical foundation for the establishment of embryo selection technology platform using metabolomics.Methods:Eight patients who received in vitro fertilization and embryo transfer (IVF-ET) treatment at Reproductive Medicine Center of Guangxi Zhuang Autonomous Region Maternal and Child Health Hospital between November 13 and December 5, 2023 were selected as the study subjects. The D3 culture medium from patient embryos was collected and divided into high-quality blastocysts ( n=42), non-high-quality blastocysts ( n=33), and embryos that failed to form blastocysts (non-formation group, n=43) according to the formation of day 5 blastocysts. High-performance liquid chromatography-mass spectrometry was employed to perform non-targeted metabolomic analysis in the D3 culture medium from three distinct groups. Results:1) The metabolites in D3 culture medium of embryos with varying developmental potentials exhibit significant differences. Specifically, 79 differential metabolites were identified between the blastocyst formation group and the non-blastocyst formation group (all P<0.05); additionally, 73 differential metabolites were found between the high-quality blastocyst group and the non-high-quality blastocyst group (all P<0.05). 2) The area under the receiver operating characteristic curve of significantly differential metabolites for predicting potential of D3 embryo blastocyst formation and high-quality blastocyst formation were both greater than 0.9, demonstrating excellent predictive performance. 3) KEGG pathway enrichment analysis revealed that differential metabolites associated with blastocyst formation potential were primarily enriched in pathways including D-amino acid metabolism, glycine-serine-threonine metabolism, arginine biosynthesis, and histidine metabolism ( P<0.05). For high-quality blastocyst formation, the differential metabolites were predominantly enriched in pathways related to tryptophan metabolism, D-amino acid metabolism, serotonergic synapses, and protein digestion and absorption ( P<0.05). Conclusion:Embryos with different developmental potentials have significantly different metabolic profiles, and it is feasible to predict the developmental potential of D3 embryos by metabolomics analysis.
3.Clinical efficacy of double incision and double plate internal fixation in the treatment of tibial plateau fractures in the elderly
Xiaobing MIAO ; Kejian HONG ; Yongping DAI ; Jun ZHANG
Journal of Clinical Surgery 2024;32(8):874-877
Objective To investigate the clinical effect of dual incision and dual plate internal fixation for the treatment of tibial plateau fractures in the elderly and its impact on knee joint function.Methods Using retrospective analysis method,103 elderly patients with tibial plateau fractures admitted to Rugao Hospital affiliated to Nantong University from April 2015 to May 2021 were selected for clinical research.According to the surgical method,they were divided into two groups,52 patients in the study group were treated with double incision double plates,and 51 patients in the control group were treated with locking plate internal fixation.Perioperative indicators,knee function,knee stability Knee joint complications.Results The hospitalization time of the study group was(7.8±2.0)days,and that of the control group was(10.0±2.4)days,and the difference was statistically significant between the two groups(P<0.05).The HSS scores at 3 months,6 months and 12 months were(66.9±5.4),(78.4±6.6)and(83.8±6.1)in the study group,and(64.2±6.1),(74.0±7.3)and(82.0±6.8)in the control group,respectively.There was statistical significance in HSS scores 3 months and 6 months after operation between the two groups(P<0.05).The tibial migration distance 3 months after operation was(2.54±0.50)mm in the study group and(2.84±0.67)mm in the control group,and the difference was statistically significant(P<0.05).The knee posterior inclination angles of the study group were(4.12±1.10)°,(5.03±0.96)° and(5.46±1.52)° at 3 months,6 months and 12 months after surgery,respectively,while those of the control group were(6.11±1.43)°,(6.67±1.54)° and(7.50±1.88)°,respectively.The difference was statistically significant(P<0.05).The activity of the study group at 3 months and 6 months after surgery was(104.3±8.2)° and(117.4±7.6)°,respectively,while that of the control group was(96.8±8.9)° and(111.8±8.2)°,and the difference was statistically significant(P<0.05).There were 3 cases of postoperative complications in the study group and 10 cases in the control group,and the difference was statistically significant(P<0.05).Conclusion The clinical effect of dual incision and dual plate internal fixation in the treatment of tibial plateau fractures in the elderly is affirmative,which is more beneficial for promoting early recovery of knee function and maintaining stability of knee function than locking plate internal fixation.
4.Investigating the method of semen treatment for the detection of sperm DNA fragmentation index
Tinglyu WEI ; Caizhu WANG ; Kejian SUN ; Zhulian WU ; Yanwen PAN ; Xianyou GAN ; Hong ZHOU ; Jinhui SHU
Chinese Journal of Reproduction and Contraception 2023;43(12):1260-1263
Objective:To investigate the effects of different semen sample collection methods on sperm DNA fragmentation index (DFI) test results, then to evaluate the accuracy of the current semen sample collection method in the assessment of male fertility.Methods:In this study, 50 semen sample obtained on the day of oocyte retrieval from patients undergoing in vitro fertilization at the Reproductive Medical Center in Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region from August 2021 to January 2022 were collected. For each semen, a small amount of samples were collected in three different retention methods for routine semen and sperm DFI testing. Three different ways of retaining samples were as follows: group A, after mixing of the semen, 50 μL sample was directly collected; group B, after density gradient centrifugation, 50 μL sample was collected at the interface between semen and gradient fluid; group C, after density gradient centrifugation, the sperm pellet was upstream, then 50 μL sample was collected from upstream liquid. After semen treatment, routine semen testing and sperm DFI testing were performed. Pearson was used to analyze the correlation between DFI and the percentage of immobile sperm and the percentage of forward sperm movement. Results:The sperm motility rate of group C [(96.83±2.28)%] was significantly higher than that of group A [(57.16±11.28)%, P<0.001] and group B [(22.54±9.35)%, P<0.001], and there was a statistical difference among the three groups. The immotile sperm rate of group B sample was (77.46±9.35)%, which was significantly higher than that of samples from group C [(3.14±2.31)%, P<0.001] and group A [(42.83±11.28)%, P<0.001]. There was also a statistical difference in DFI among the three groups ( P<0.001). The DFI of group B [37.18% (30.41%, 47.80%)] was significantly higher than that of group A [22.00% (14.75%, 29.25%), P<0.001] and group C [0.78% (0.00%, 2.07%), P<0.001]. Pearson analysis results showed that the DFI of group A and group B was positively correlated with the percentage of immobile sperm ( r=0.304, P=0.032; r=0.612, P<0.001), while the DFI of group B was negatively correlated with the percentage of sperm forward movement ( r=-0.517, P<0.001). Conclusion:For the same semen, the DFI of immotile sperm was significantly higher than that of motile sperm. Therefore, due to the interference of immotile sperm, the DFI value by the current sample retention method cannot accurately reflect the DNA status of active sperm participating in fertilization. This suggests that the samples used for DFI testing should be collected from motile sperm collected by gradient centrifugation, upstream or other methods, which can more accurately assess male fertility.
5.Investigating the method of semen treatment for the detection of sperm DNA fragmentation index
Tinglyu WEI ; Caizhu WANG ; Kejian SUN ; Zhulian WU ; Yanwen PAN ; Xianyou GAN ; Hong ZHOU ; Jinhui SHU
Chinese Journal of Reproduction and Contraception 2023;43(12):1260-1263
Objective:To investigate the effects of different semen sample collection methods on sperm DNA fragmentation index (DFI) test results, then to evaluate the accuracy of the current semen sample collection method in the assessment of male fertility.Methods:In this study, 50 semen sample obtained on the day of oocyte retrieval from patients undergoing in vitro fertilization at the Reproductive Medical Center in Maternity and Child Health Hospital of Guangxi Zhuang Autonomous Region from August 2021 to January 2022 were collected. For each semen, a small amount of samples were collected in three different retention methods for routine semen and sperm DFI testing. Three different ways of retaining samples were as follows: group A, after mixing of the semen, 50 μL sample was directly collected; group B, after density gradient centrifugation, 50 μL sample was collected at the interface between semen and gradient fluid; group C, after density gradient centrifugation, the sperm pellet was upstream, then 50 μL sample was collected from upstream liquid. After semen treatment, routine semen testing and sperm DFI testing were performed. Pearson was used to analyze the correlation between DFI and the percentage of immobile sperm and the percentage of forward sperm movement. Results:The sperm motility rate of group C [(96.83±2.28)%] was significantly higher than that of group A [(57.16±11.28)%, P<0.001] and group B [(22.54±9.35)%, P<0.001], and there was a statistical difference among the three groups. The immotile sperm rate of group B sample was (77.46±9.35)%, which was significantly higher than that of samples from group C [(3.14±2.31)%, P<0.001] and group A [(42.83±11.28)%, P<0.001]. There was also a statistical difference in DFI among the three groups ( P<0.001). The DFI of group B [37.18% (30.41%, 47.80%)] was significantly higher than that of group A [22.00% (14.75%, 29.25%), P<0.001] and group C [0.78% (0.00%, 2.07%), P<0.001]. Pearson analysis results showed that the DFI of group A and group B was positively correlated with the percentage of immobile sperm ( r=0.304, P=0.032; r=0.612, P<0.001), while the DFI of group B was negatively correlated with the percentage of sperm forward movement ( r=-0.517, P<0.001). Conclusion:For the same semen, the DFI of immotile sperm was significantly higher than that of motile sperm. Therefore, due to the interference of immotile sperm, the DFI value by the current sample retention method cannot accurately reflect the DNA status of active sperm participating in fertilization. This suggests that the samples used for DFI testing should be collected from motile sperm collected by gradient centrifugation, upstream or other methods, which can more accurately assess male fertility.
6.Improvement of Quality Standard for Mongolian Medicine Artemisia sacrorum
Xiaohong YUAN ; Kejian PANG ; Hui TANG ; Hong ZHANG ; Feng WEI ; Lele LIU ; Guozhen JIANG
China Pharmacy 2021;32(5):536-541
OBJECTIVE:To improve the quality standard of M ongolian med icine Artemisia sacrorum ,and to provide scientific basis for comprehensive quality evaluation. METHODS :The appearance and microscopic characteristics of A. sacrorum were identified;scopoletin,chlorogenic acid ,caffeic acid ,scopoletin and 3,5-dicaffeoylquinic acid were identified quantitatively by TLC;the contents of above 5 components were determined by HPLC. The water content ,total ash and extract were examined. RESULTS:The stem of A. sacrorum was cylindrical ,and its surface was purple or purple-brown or cyan-brown ;the leaves were ovate or oblong-ovate ,fragrant;the flowers were yellow ,head-shaped,subglobose or hemispherical. The powder was green or yellow-green,its pollen grain had three germination ;the parenchymal cell clusters with sharp edges and numerous threaded ducts , occasionally having marginal pitted ducts ;its wood fibers were in bundles mostly. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for 5 substance control and samples. The linear range of scopoletin, chlorogenic acid , caffeic acid , scopolactone and 3,5-dicaffeoylquinic acid were 85.60-428.00, 10.16-101.60, 10.20-102.00,40.84-408.40 and 40.80-408.00 μg/mL(all r>0.999 0). RSDs of precision ,stability,repeatability tests were all less than 3.00%(n=6). The average recoveries were 103.07%,99.66%,98.37%,97.78%,98.40%(all RSDs <3.00%,n=6). The contents of the above-mentioned 5 compounds in 10 batches of samples were 0.36%-1.23%,0.09%-0.51%,0.04%-0.13%, 0.61% -1.13% ,0.12% -1.11% ,respectively;the average com contents of water ,total ash and water soluble extract were 6.25%,5.86%,26.50%,respectively. CONCLU SIONS:O the basis of the original quality standard of A. sacrorum , microscopic identification,TLC identification ,content determination and examination items of water ,total ash and extract are added. The method shows good precision ,accuracy and stability ,which can provide reference for more scientific and standardized evaluation of the quality of this medicinal material.
7.Improvement of Quality Standard for Mongolian Medicine Juniperus rigida
Xiaohong YUAN ; Kejian PANG ; Hui TANG ; Hong ZHANG ; Hui GUAN ; Feng WEI ; Ping TANG
China Pharmacy 2021;32(16):1949-1955
OBJECTIVE:To provide scientifi c evidence for improving the quality standard of Mongolian medicine Juniperus rigida. METHODS :Totally 10 batches of J. rigida from different places were taken as samples to observe their characters and identify them by microscope ;TLC method was adopted to qualitatively identify isoquercitrin ,quercitrin,amentoflavone, podocarpusflavone A and hinokiflavone ;the contents of total ash ,acid-insoluble ash ,ethanol-soluble extract and heavy metals were determined by related method stated in 2020 edition of Chinese Pharmacopeia (part Ⅳ). The contents of above 5 components in samples were determined by HPLC. RESULTS :The powder of J. rigida was green or yellowish green ,polygonal tracheids , closely arranged in longitudinal with unequal stomatal ;epidermal cells were nearly rectangular ;sclerenchyma cells were quasi rectangular and the wall beadedly thickening. Results of TLC showed that the spots of the same color were found in the corresponding positions of chromatogram for test sample and substance control. The contents of total ash ,acid-insoluble ash and ethanol-soluble extract in 10 batches of samples were 7.37%-11.18%,0.75%-2.98%,16.55%-26.42%,respectively;average contents were 8.51%,1.27%,22.35%. The contents of lead ,arsenic,cadmium,mercury and copper were 2.00-5.44,0.65-1.65, 0.044-0.100,0.034-0.160,4.59-6.79 mg/kg,respectively;average conte nts were 3.73,0.97,0.078,0.061,5.23 mg/kg. The linear ranges of isoquercitrin ,quercitrin,amentoflavone,podocarpus- flavone A and hinokiflavone were 4.98-20.02,49.99-199.96, 19.94-99.96,9.99-40.00,20.20-159.98 μg/mL(all r>0.999 7); com RSDs of precision ,repeatability and stability (24 h) tests were all less than 3.00%(n=6);the average recoveries were 话:0993-2057878。E-mail:Tanghuishz@qq.com 100.62%-102.96%,RSDs were 1.21%-1.88%(n=6). Average contents of the above-mentioned 5 compounds in 10 batches of samples were 0.089-0.379,1.379-4.250,1.077-2.026,0.162-0.423, 0.016 9-0.117 0 mg/g,respectively. CONCLUSIONS :The qualitative and quantitative analysis methods of Mongolian medicine J. rigida are established. It is preliminarily proposed that the total ash content shall not exceed 10.22%,the acid-insoluble ash content shall not exceed 1.53%,ethanol-soluble extract content shall not be less than 17.88%,heavy metal lead should not exceed 5 mg/kg,arsenic should not exceed 2 mg/kg,cadmium should not exceed 0.3 mg/kg,mercury should not exceed 0.2 mg/kg,copper should not exceed 20 mg/kg.
8.Effects of CXCR7-shRNA lentiviral vector on the growth and invasiveness of human hepatoma carcinoma cells in vitro.
Xiaozhen DAI ; Xin XIONG ; Lan WANG ; Kejian PAN ; Lang HE ; Hong LI
Journal of Southern Medical University 2013;33(7):994-998
OBJECTIVETo explore the effects of CXCR7 knock-down by CXCR7-shRNA lentiviral vector on the proliferation and invasion of human hepatoma carcinoma cells in vitro.
METHODSCXCR7-shRNA lentiviral vector was transfected into hepatocellular carcinoma HCCLM3 cells. The changes in mRNA and protein expression of CXCR7 in the transfected cells were investigated using real-time PCR and Western blotting, respectively, and MTT assay was employed to assess the cell proliferation changes. In vitro adhesion assay and transwell chamber test were used to observe the adhesion and invasiveness of HCCLM3 cells, respectively.
RESULTSTransfection of HCCLM3 cells with CXCR7-shRNA lentiviral vector resulted in a significantly decreased expression of CXCR7 at both mRNA and protein levels (P<0.01) and obvious suppression of the cell proliferative activity (P<0.05). CXCR7-shRNA also significantly suppressed the invasiveness and adhesion of HCCLM3 cells (P<0.01).
CONCLUSIONCXCR7 knock-down can significantly inhibit the proliferation and invasiveness of human hepatoma carcinoma cells in vitro, suggesting the value of CXCR7 as a potential target for hepatoma carcinoma therapy.
Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Liver Neoplasms ; genetics ; pathology ; RNA, Small Interfering ; genetics ; Receptors, CXCR ; genetics ; Transfection
9.Application of OMIM in medical genetics teaching and its reflection
Ya LI ; Kejian PAN ; Lan WANG ; Lang HE ; Hong LI ; Xiaozhen DAI ; Yuhan YANG
Chinese Journal of Medical Education Research 2013;(3):233-235
With the rapid development of medical genetics,online Medelian inheritance in man (OMIM) manifests a more and more important role in medical genetics teaching.Using the educational form combining ‘ classroom teaching,review writing and seminar’,‘ Query and use of OMIM ’was introduced into the education of medical genetics.Reality practice revealed that this educational practice maintained advanced and timely status of knowledge and deeply activated self-studying and independent thinking ability of students.
10.Evaluation of cardiopulmonary allograft function for a combined heart-lung transplantation patient survived 5 years
Shouguo YANG ; Chunsheng WANG ; Hao CHEN ; Shijie ZHU ; Ying ZHANG ; Tao HONG ; Yamin ZHUANG ; Kejian HU
Fudan University Journal of Medical Sciences 2010;37(1):88-91
Objective To evaluate the cardiopulmonary allograft function and to analyze key factors for long-term survival of heart-lung transplantation in a patient survived more than 5 years. Methods On December 17th, 2003 at Zhongshan Hospital of Fudan University, a homologous heart-lung transplantation was performed on a female who diagnosed with cardiopulmonary failure secondary to congenital atrial septal defect with severe pulmonary hypertension. Heart-lung allograft was preserved with 1 000 mL UW solution and 4 000 mL HTK solution.Postoperative immunosuppressive therapies were managed with Zenapax, cyclosporine A (or tacrolimus), mycophenolate mofetil and corticosteroids. Cyclosporine A maintained with serum trough levels of 100-200 μg/L and tacrolimus with serum trough levels of 8-20 μg/L. Cardiopulmonary allograft functions were evaluated by echocardiogram, pulmonary function test and thoracic CT periodically. Results The patient survived operation and experienced normal daily life with NYHA cardiac function of class Ⅰ-Ⅱ during the follow-up of 5 years and 6 months. Echocardiogram showed left ventricular ejection fraction of 65% to 86%. Pulmonary function test exhibited with nearly normal oxygen exchange, meanwhile, small airway obstruction was detected from one year after operation and keeping stable from then on. Two episodes of severe pneumonia were complicated and treated with antibiotics and fhconazob, no severe acute allograft rejection episode was experienced. Conclusions Heart-lung transplantation proves to be a reliable therapy modality for terminal cardiopulmonary failure. Excellent donor organ preservation, accurate balance of the risk between acute allograft rejection and infection, and strict preventive measures against infection are key factors for long-term survival of heart-lung transplantation.

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