1.Biological characterization of C57 mouse bone marrow mesenchymal stem cells using a whole bone marrow adherent culture technique
Rongyao ZHANG ; Xiaojuan BI ; Yan MA ; Xianlin DUAN ; Ming JIANG
Chinese Journal of Tissue Engineering Research 2014;(1):45-50
BACKGROUND:Under mitomycin C treatment, feeder cells appear to have restricted proliferation, but they are stil able to secret different cytokines. Non-mesenchymal stem cells from the bone marrow and secreted factors in plasma maintain the micro-environment suitable for the growth of mesenchymal stem cells that can improve the yield of mesenchymal stem cells.
OBJECTIVE:To study the biological characteristics of C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique.
METHODS:Using the whole bone marrow adherent culture technique, purified and amplified C57 mouse bone marrow mesenchymal stem cells were harvested. cellproliferation kinetics, immune cellsurface markers, multiple differentiation potential and cellcycle were detected.
RESULTS AND CONCLUSION:Using the whole bone marrow culture, mouse bone marrow mesenchymal stem cells were harvested and capable of adhering to the plastic culture vessel. The obtained cells expressed CD45, CD105 and Sca-1, but were negative for CD34, CD33 and C-kit. The doubling time was (57.11±1.5) hours. The cells could be induced to differentiate into osteoblasts, adipocytes and chondrocytes. The cellcycle analysis showed that 64%of cells were in G 0-G 1 phase. These indicates that C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique have biological characteristics of mesenchymal stem cells.
2.The diagnostic value of the change of different parts of electrocardiogram in treadmill exercise test for coronary artery disease
Jianling SUN ; Weimin HUANG ; Congyu WANG ; Jihong GUO ; Xiaoying LI ; Shutian SHI ; Xianlin MA ; Juan WANG
Clinical Medicine of China 2012;28(11):1177-1180
Objective To determine the diagnostic value of the change of different parts of electrocardiogram in treadmill exercise test (TET) for coronary heart disease (CHD).MethodsFrom Jan.2006 to Mar.2011,445 patients with CHD underwent treadmill exercise test and coronary angiography (CAG) in our hospital.We analyzed retrospectively the diagnostic value of the change of different parts of electrocardiogram in TET test.Results ( 1 ) There were 200 cases who had positive results during treadmill exercise test and 150 cases of them had been diagnosed of CHD by coronary angiography;The other 245 cases had negative results during treadmill exercise test and 39 cases of them had been diagnosed as CHD by coronary angiography.The sensitivity and specificity rates of treadmill exercise test in diagnosis of CHD were 79.36%( 150/189 ) and 80.47% ( 206/256 ) respectively. ( 2 ) Of the 200 positive cases tested by coronary angiography,150 cases were diagnosed of CHD,including 22,58 and 70 cases with limb,chest and combined chest and limb leads positive results respectively.The positive rates for the groups of combined chest and limb leads and limb ST leads were significantly higher than that of the chest leads only group ( x2:6.34,3.93 ; P <0.05).(3)R wave amplitude of the CAG negative group after exercise (17.54 ± 2.52)mm was significantly lower than the CAG positive group ( 19.42 ± 3.46 )mm ( t =6.33,P < 0.05 ).( 4 ) For the 200 cases with positive TET,there were no significant statistical difference ( P > 0.05 ) on CAG positive rate between T wave normalization group [ 73.3% (44/60) ] with and TET positive group [ 75.0% (105/140) ].But these two groups had significantly higher ( x2:80.21,132.82 ; P < 0.05 ) CAG positive rates than TET negative group [ 15.9% (39/245) ].ConclusionThe treadmill exercise test is valuable in noninvasive diagnosis of CHD.The severity of CAG can be preliminarily estimated by analyzing the pattern of the change of different parts of electrocardiogram
3.Human umbilical cord mesenchymal stem cells:Comparison of hematopoiesis supporting capacity before and after cryopreservation
Xiangping CHANG ; Yan MA ; Xiaojuan BI ; Lijuan SONG ; Xianlin DUAN ; Ming JIANG
Chinese Journal of Tissue Engineering Research 2013;(32):5765-5771
BACKGROUND:Cryopreservation of human umbilical cord mesenchymal stem cel s has been a hot research issue currently, but the studies concerning their effects on expansion of hematopoietic stem/progenitor cel s after cryopreservation are seldom. OBJECTIVE:To investigate the effects of human umbilical cord mesenchymal stem cel s before and after cryopreservation as feeder layer on expansion of human bone marrow mononuclear cel s in vitro. METHODS:2.5g/L mitomycin C processed human umbilical cord mesenchymal stem cel s and bone marrow mesenchymal stem cel s at passage 3 were used as the feeder layer to expand adult al ogeneic bone marrow mononuclear cel s in culture. Up to day 35, methylcel ulose assay was used to detect hematopoietic stem/progenitor cel colony proliferation. RESULTS AND CONCLUSION:There were no differences in the morphology and size of colonies in the cryopreserved human umbilical cord mesenchymal stem cel group, bone marrow mesenchymal stem cel group and non-cryopreserved human umbilical cord mesenchymal stem cel group. However, these parameter described above were significantly higher in these three groups than the blank control group (P<0.05). There were fewer colonies in the cryopreserved human umbilical cord mesenchymal stem cel group than the non-cryopreserved human umbilical cord mesenchymal stem cel group (P<0.05). These findings indicate that human umbilical cord mesenchymal stem cel s before and after cryopreservation have the ability as feeder layer on expansion of bone marrow mononuclear cel s in vitro similar to bone marrow mesenchymal stem cel s. But this ability of human umbilical cord mesenchymal stem cel s may decrease after cryopreservation.
4.Application of reserved jejunal feeding tube during postoperative chemotherapy of gastric cancer
Hanjun MA ; Sanrong LI ; Hansheng JIN ; Jian SUN ; Xianlin SHU ; Yalin YE ; Qingquan LV ; Qi WEI
Chinese Journal of Postgraduates of Medicine 2008;31(20):30-32
Objective To evaluate the effects of reserved jejunal feeding tube during postoperative chemotherapy of gastric cancer. Methods Forty-two patients with gastric cancer underwent radical gastrectomy and going to adjuvant chemotherapy,conventional placed jejunal feeding tube. All of the patients weredivided into group A and group B randomly by pathological staging and tumor site, group A reserved jejunal feeding tube and received enteral nutrition through the tube during chemotherapy, and group B non-reservedjejunal feeding tube and been given daily diet,compared nutrition and immune indicators of two groups beforeand after chemotherapy ,compared the rate of vomiting,and observed complications long-term reserved jejunal feeding tube. Results In post-chemotherapy,nutrition and immune indicators of group A were betterthan those of group B, the difference was statistically significant (P<0.05) ,the rate of vomiting in group Awas significantly lower than that of in group B (X2= 9.75, P<0.01 ), no serious complieations occurred forlong-term reserved jejunal feeding tube. Conclusions Reserved jejunal feeding tube and received enteralnutrition through the tube during postoperative chemotherapy of gastric cancer can significantly improve the nutritional and immune status. It is safe and reliable, worth promoting.
5.A Novel EYA1 Mutation Causing Alternative RNA Splicing in a Chinese Family With Branchio-Oto Syndrome: Implications for Molecular Diagnosis and Clinical Application
Anhai CHEN ; Jie LING ; Xin PENG ; Xianlin LIU ; Shuang MAO ; Yongjia CHEN ; Mengyao QIN ; Shuai ZHANG ; Yijiang BAI ; Jian SONG ; Zhili FENG ; Lu MA ; Dinghua HE ; Lingyun MEI ; Chufeng HE ; Yong FENG
Clinical and Experimental Otorhinolaryngology 2023;16(4):342-358
Objectives:
. Branchio-oto syndrome (BOS) primarily manifests as hearing loss, preauricular pits, and branchial defects. EYA1 is the most common pathogenic gene, and splicing mutations account for a substantial proportion of cases. However, few studies have addressed the structural changes in the protein caused by splicing mutations and potential pathogenic factors, and several studies have shown that middle-ear surgery has limited effectiveness in improving hearing in these patients. BOS has also been relatively infrequently reported in the Chinese population. This study explored the genetic etiology in the family of a proband with BOS and provided clinical treatment to improve the patient’s hearing.
Methods:
. We collected detailed clinical features and peripheral blood samples from the patients and unaffected individuals within the family. Pathogenic mutations were identified by whole-exome sequencing and cosegregation analysis and classified according to the American College of Medical Genetics and Genomics guidelines. Alternative splicing was verified through a minigene assay. The predicted three-dimensional protein structure and biochemical experiments were used to investigate the pathogenicity of the mutation. The proband underwent middle-ear surgery and was followed up at 1 month and 6 months postoperatively to monitor auditory improvement.
Results:
. A novel heterozygous EYA1 splicing variant (c.1050+4 A>C) was identified and classified as pathogenic (PVS1(RNA), PM2, PP1). Skipping of exon 11 of the EYA1 pre-mRNA was confirmed using a minigene assay. This mutation may impair EYA1-SIX1 interactions, as shown by an immunoprecipitation assay. The EYA1-Mut protein exhibited cellular mislocalization and decreased protein expression in cytological experiments. Middle-ear surgery significantly improved hearing loss caused by bone-conduction abnormalities in the proband.
Conclusion
. We reported a novel splicing variant of EYA1 in a Chinese family with BOS and revealed the potential molecular pathogenic mechanism. The significant hearing improvement observed in the proband after middle-ear surgery provides a reference for auditory rehabilitation in similar patients.
6.The application of ultrasound in the diagnosis of small bowel volvulus in adults
Yanwen LUO ; Qingli ZHU ; Wenbo LI ; Wei LIU ; Lin CONG ; Xianlin HAN ; Yingxin WEI ; Guannan ZHANG ; Mengsu XIAO ; Li MA
Chinese Journal of Surgery 2023;61(10):907-912
Objective:To analyze the ultrasound characteristics of small bowel volvulus among adults and to investigate the value of ultrasound in the diagnosis of small bowel volvulus.Methods:Totally 34 adults with small bowel volvulus confirmed by clinical diagnosis or surgery and who underwent ultrasound examination at Peking Union Medical College Hospital from August 2017 to October 2022 were enrolled, including 19 males and 15 females, aged (55.0±21.8) years (range: 19 to 94 years). The clinical characteristics, CT images and ultrasound images of the patients were retrospectively reviewed, and the ultra, sound features of small bowel volvulus and its diagnostic efficacy were analyzed.Results:Abdominal pain was the typical clinical symptom of all patients. Other symptoms included 21 cases of abdominal distension, 19 cases of nausea and vomiting, and 13 cases of cessation of passage of stool or flatus. Eight patients had signs of peritonitis and 22 patients had abnormal bowel sounds. Twenty patients had a history of abdominal surgery. Twenty-seven patients underwent surgery for intestinal obstruction, and the remaining 7 patients improved after conservative treatment. All cases were evaluated by ultrasound, 11 cases showed a “whirl sign” and were diagnosed as small bowel volvulus, the diagnostic accuracy rate was 32.4% (11/34), ultrasound simultaneously diagnosed intestinal obstruction in 21 cases, 17 cases of abdominal effusion, 4 cases of intestinal wall thickening, 2 cases of abdominal mass, 1 case of intussusception, 1 case of right sided inguinal hernia. CT and ultrasound had a consistent positive discovery in 88.2% (30/34) of all the patients.Conclusion:Ultrasound is valuable in the diagnosis of small bowel volvulus and the evaluation of complications.
7.The application of ultrasound in the diagnosis of small bowel volvulus in adults
Yanwen LUO ; Qingli ZHU ; Wenbo LI ; Wei LIU ; Lin CONG ; Xianlin HAN ; Yingxin WEI ; Guannan ZHANG ; Mengsu XIAO ; Li MA
Chinese Journal of Surgery 2023;61(10):907-912
Objective:To analyze the ultrasound characteristics of small bowel volvulus among adults and to investigate the value of ultrasound in the diagnosis of small bowel volvulus.Methods:Totally 34 adults with small bowel volvulus confirmed by clinical diagnosis or surgery and who underwent ultrasound examination at Peking Union Medical College Hospital from August 2017 to October 2022 were enrolled, including 19 males and 15 females, aged (55.0±21.8) years (range: 19 to 94 years). The clinical characteristics, CT images and ultrasound images of the patients were retrospectively reviewed, and the ultra, sound features of small bowel volvulus and its diagnostic efficacy were analyzed.Results:Abdominal pain was the typical clinical symptom of all patients. Other symptoms included 21 cases of abdominal distension, 19 cases of nausea and vomiting, and 13 cases of cessation of passage of stool or flatus. Eight patients had signs of peritonitis and 22 patients had abnormal bowel sounds. Twenty patients had a history of abdominal surgery. Twenty-seven patients underwent surgery for intestinal obstruction, and the remaining 7 patients improved after conservative treatment. All cases were evaluated by ultrasound, 11 cases showed a “whirl sign” and were diagnosed as small bowel volvulus, the diagnostic accuracy rate was 32.4% (11/34), ultrasound simultaneously diagnosed intestinal obstruction in 21 cases, 17 cases of abdominal effusion, 4 cases of intestinal wall thickening, 2 cases of abdominal mass, 1 case of intussusception, 1 case of right sided inguinal hernia. CT and ultrasound had a consistent positive discovery in 88.2% (30/34) of all the patients.Conclusion:Ultrasound is valuable in the diagnosis of small bowel volvulus and the evaluation of complications.