1.Congenital hypothyroidism characterized by hydrops fetalis:one case report
Yan JIANG ; Haiqing LENG ; Jun BU
Journal of Clinical Pediatrics 2016;34(9):664-666
Objective To explore the differential diagnosis of hydrops fetalis and the rare presentations of neonatal congenital hypothyroidism. Methods The data of one congenital hypothyroidism diagnosed neonate with hydrops fetalis leading to birth asphyxia and respiratory failure were retrospectively analyzed. The relevant literatures were reviewed. Results A Uyghur female infant by cesarean delivery at gestational age of 38+5 week for intrauterine distress, presented general edema with cyanosis and dyspnea after birth. Trachea cannula was used to assist ventilation. At one-day old, the thyroid function examination showed that the serum thyroid stimulating hormone was>100 mU/L and the free thyroid was 6 . 56 pmol/L. Moreover, ultrasonographic examination indicated the thyroid aplasia. The clinical symptoms were improved after the treatment with the levothyroxine tablets replacement, and breathing machine was removed at 8-day old. The dosage of drug was adjusted by clinical manifestation and laboratory monitoring. The patient was discharged at 18-day old with the medicine and was followed-up. Conclusions Congenital hypothyroidism can be the pathogenesis of hydrops fetalis and its differential diagnosis should be paid attention.
2.Stable expression of mouse IFN-λ2 in CHO cells and its biological activity analysis
Yulan YAN ; Lixue YUAN ; Yang LIU ; Wenyan CAO ; Xuefeng BU ; Zhigao BU ; Jinxu ZHENG
Chinese Journal of Microbiology and Immunology 2010;30(2):104-109
Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.
3.A novel scaffold for endothelial progenitor cells in tissue engineered vascular grafts
Xuefeng BU ; Yulan YAN ; Zhijian ZHANG ; Haorong WU
Chinese Journal of General Surgery 2009;24(1):34-37
Objective To explore the characteristics of endothelial progenitor cells (EPCs) on different scaffolds and to find a new bio-engineered synthetic hybrid scaffold for artificial bio-engineered blood vessels. Methods EPCs were induced from mesenchymal stem cells isolated from rat bone marrow and seeded on ECM scaffold. The surface structure of the scaffold and growth status of EPCs on the scaffold were observed and analysed by electron microscopy. The characteristics and number of those EPCs on different kinds of scaffolds were studied with EPC-specific VWF by immunofluorescence, Western blotting and real-time PCR technique at different time points. Results The cell adhesion rate at 1,3,5 h after seeded on pressed scaffold were higher than that on unpressed scaffolds( P < 0. 01 ). Pressed scaffolds has got a larger cell number( P < 0. 05 )at DIV1, DIV3, DIV7, but there was no significant difference after DIV10. Furthermore, cell shapes of EPCs on pressed scaffolds were more mature and more similar to endothelial cells. A level cell surface on pressed scaffolds was achieved. Western blotting assays revealed EPCs on pressed scaffolds expressed more protein VWF at DIV3, DIV7, DIV10. Real-time PCR results showed EPCs on the two different groups of scaffolds all expressed VWF gene, The quantity of their expression in the two groups were all enhanced after DIV7 (P < 0. 05 ). The quantity of VWF gene expression in the pressed group was much higher than that in the unpressed group at DIV3 ,DIV7,DIV10 (P <0, 01), but there was no significant difference after DIV14. Conclusions Pressed ECM scaffolds can promote adhesion, proliferation and differentiation on EPCs. Pressed scaffolds can be used as the matrix for EPC and fabricated into a novel synthetic tissue bio-engineered vascular scaffold.
4.Volumetric and dosimetric variations in radiation treatment planning using CT images based on different reconstructed field-of-view
Zhongjie LU ; Senxiang YAN ; Luyi BU ; Jinqi ZHOU
Chinese Journal of Radiological Medicine and Protection 2010;30(3):303-306
Objective To analyze the volumetric and dosimetric variations in radiation treatment planning(RTP) using CT images based on normal and extended reconstructed field-of-view(FOV). Methods Original data of CT scans from 16 cases of nasopharyngeal carcinomas were reconstructed to form 2 sets of CT images with Dermal(45 cm)and EFOV(65 cm),which were then exposed to RTP. Contouring of targets/OAR including GTV(gross tumor volume),CTV(clinical target volume,CTV),brain stem, lens, parotids and cord was made on normsl FOV CT set.A 7-field equi-angular IMRT (intensity modulated radiation Therapy)plan was generated with prescribed GTV dose of 70 Gy.Two sets 0f CT images were fused in DICOM coordinate system and targets/OARs on normal FOV CT were copied to EFOV CT.IMRT plans were then transplanted from normal FOV to EFOV CT,with the same isocenter on DICOM coordinates.Volumetric and dosimetrie variations including GTV,CTV brain stem,lens, parotids and cord were calculated on dose-volume-histogram(DVH).For dosimetric verification,IMRT plans were input into fluence maps of Mapcheck 1175 phantom based on normal FOV and EFOV, and DTA(distance to agreement)was used to analyze the passing rate of calculated/measured absolute doses at 5 cm depth.Paired-t test was used to compare the passing rate of field 1-7 of IMRT plans based on 2 CT sets.Results Volumes of targets and OARs on 2 CT sets of different FOVs were statistically different.with larger calculated volume on norlual FOV in all cases.There was no statistic difference in the maximal(Dmax) doses received by all targets and OARs except the small-volume lens, in which the dose was higher on normal CT than that on EFOV CT(t=-3.14,P<0.007).The mean doses(Dmean)to the CTV(clinical target volume)and GTV(gross tumor volume)were higher on EFOV than normal FOV CT(t=-6.45,-5.65,P<0.001).There was no statistic difference in Dmean received by OARs and the minimal dose (Dmin)by all targets and OARs(P>0.05).There was also no statistic difference in the passing rate of field 1-7 of IMRT plans based on 2 CT sets.Conclusions There were volumetric and dosimetric variations as evaluated on DVH using different reconstructed FOV during CT simulation,though the difference between the passing rates as verified in 2 dimensional fluence map was not significant.
5.Application value of MDCT double period enhanced scanning with low-tension water enteroclysis pres-entations for colon cancer preoperative staging
Jie ZHANG ; Qingfeng BU ; Fuwen FU ; Ping HU ; Guanghui YAN
Practical Oncology Journal 2013;(6):500-503
Objective This paper is mainly to discuss accuracy and clinical application value of MDCT double-period enhanced scanning with low -tension water enteroclysis for colon cancer preoperative TNM stag-ing.Methods Sixty-two colon cancer patients with complete images and pathological data were selected in our hospital from January 2012 to May 2013 .We retrospectively analyzed CT image changes of the tumor location ,the extent of tumor invasion,the surrounding fat space,lymph node metastasis and distant metastasis.We compared them with postoperative pathology to prove the accuracy of MDCT double -period enhanced scanning with low -tension water enteroclysis.Results The results showed that its accuracy rate reached to 90.32%(56/62)in co-lon cancer preoperative Stage T,80.64%(50/62)in Stage N,and 100%(62/62)in Stage M respectively.Con-clusions MDCT double-period enhanced scanning with low -tension water enteroclysis can accurately display the site of colon cancer and determine the scope of tumor invasion ,lymph node metastasis and distant metastasis , and give more precise diagnosis of colon cancer and preoperative staging assessments .In conclusion , it can be used as the preferable method of preoperative examination in the colon cancer .
6.Culture and identification of smooth muscle progenitor cells from rat bone marrow
Xuefeng BU ; Yulan YAN ; Yang LIU ; Zhijian ZHANG ; Mubin WANG
Basic & Clinical Medicine 2006;0(11):-
Objective To isolate and identify smooth muscle progenitor cells(SPCs) from rat bone marrow and to observe specific expression of smooth muscle progenitor cells during proliferation and differentiation in vitro.Methods MNCs were isolated by density gradient centrifugation from rat marrow and cultured in conditioned nutrient medium,identification was performed by immunofluorescent staining(?-SMA,CD14).Smooth muscle cells specific markers(?-SMA) were checked with Western blotting and Real-time PCR at different time.Results During culturing,cells adhered and became spindle shaped with outgrowth at 4 d and 7 d,and showed typical "peak" "valley" at 14 d.Both ?-SMA and CD14 were positive after 4 d.Expression of ?-SMA was not found at 1 d with Western blotting,but it gradually enhanced at 4 d and reached the peak from 10 d to 14 d,then maintaned high-level at 21 d.The results with Real-time PCR indicated that no expression of ?-SMA mRNA within non-induced cells was found,but after being induced it gradually enhanced at 4 d and got to peak at 14 d,then kept the high-level at 21 d,low-expression at 1 d was significantly different from the other ones(P
7.Quantitative detection of methylation status in the multi-drug resistance 1 gene by competitive polymerase chain reaction combined with restrictive endonucleases
Yan ZHU ; Shulan WU ; Chenxue QU ; Dingfang BU
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To establish a method of competitive polymerase chain reaction combined with restrictive endonucleases to measure the methylation quantitatively in MDR1 promoter region.Methods One sense primer and two anti-sense primers were designed to amplify a fragment of MDR1 gene promoter region, which was located between -102 and +186 bp containing a methylation site. The internal reference DNA fragment, witch was less 30bp than target fragment was made by three times of polymerase chain reaction(PCR)using different anti-sense primer and then subcloned into the Pbluescriptsk+ plasmid. The genomic DNA digested by Hpa, a methylation-sensitive restrictive endonuclease and competitive internal reference DNA were competitively amplified for MDR1 promoter in the same tube by PCR. The PCR products were electrophoresed on agarose gel, stained by ethidium bromide and subjected to image analysis scanner. The amount of target fragment was calculated as following, the optical density ratio of target fragment to competitive internal reference fragment multiplied the amount of competitive internal reference DNA. The ratio of PCR products amplified from HpaⅡ digested DNA and undigested DNA was named the methylation rate.Results The genomic DNA serially diluted with optimal amount of competitive internal reference DNA were co-amplified for MDR1 promoter. The significant positive correlation between the ratio of two products and the amount of genomic DNA was demonstrated. The correlation coefficient was 0.992, P
8.Observation on therapeutic effect of acupuncture at Shiqizhui (Extra) for primary dysmenorrhea at different time.
Chinese Acupuncture & Moxibustion 2011;31(2):110-112
OBJECTIVETo explore the best treatment opportunity of acupuncture at Shiqizhui (Extra) for treating primary dysmenorrhea.
METHODSEighty cases with primary dysmenorrhea were randomly divided into an acupuncture at Shiqizhui (Extra) in the premenstrual period group (group A, n = 20), an acupuncture at Shiqizhui (Extra) when pain occurs group (group B, n = 20) and a blank group (group C, n = 40). Both of acupuncture groups were treated and followed up for 3 consecutive menstrual cycles, respectively. The therapeutic effects were compared by use of the Cox Menstrual Symptom Scale (CMSS).
RESULTSThere were no significant differences in CMSS between the two acupuncture groups during each course and the follow-up period (all P > 0.05), but the CMSS in the two acupuncture groups were all obviously lower than those in the blank group during the same period (P < 0.05, P < 0.01).
CONCLUSIONAcupuncture at Shiqizhui (Extra) has certain therapeutic effect on primary dysmenorrhea either in the premenstrual period or when pain occurs and there is no significant difference in acupuncture at different time.
Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Dysmenorrhea ; therapy ; Female ; Humans ; Time Factors ; Young Adult
9.Effect of PaCO_2 Modulating During Operation on Post-Operative Cognitive Function of Patients Undergoing OPCAB
fu-jun, ZHANG ; yan, LUO ; bu-wei, YU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(03):-
Objective To observe the effect of PaCO_2 modulating during operation on post-operative cognitive function of patients undergoing off-pump coronary artery(OPCAB). Methods Thirty patients undergoing OPCAB were randomly divided into traditional group (G_ 1 , n=15) and modulated group (G_ 2 , n=15). During operation, PaCO_ 2 in G_ 1 maintained 35 mmHg to 39 mmHg with relatively fixed ventilation parameters setting, and PaCO_ 2 in G_ 2 ranged from 40 mmHg to 45 mmHg by adjusting ventilation parameters. Continuous cardiac output index (CCI), SvO_ 2 , regional cerebral O_ 2 saturation (rSO_ 2 ) and PaCO_ 2 were recorded before distal anastomosis(T_ 1 ), at 5 min of the first distal vessel anastomosis(T_ 2 ), second distal vessel anastomosis(T_ 3 ) and third distal vessel anastomosis(T_ 4 ), and 20 min after the completion of coronary artery anastomoses. HDS-R and ADL were used to examine the patients' cognitive function. Results There were no significant differences in pre- and post-operative HDS-R and pre-operative ADL scores between groups. The score of post-HDS-R in G_ 1 was obviously lower than that of pre-HDS-R (P
10.Activation of PPAR-γ inhibits differentiation of rat osteoblasts by reducing expression of connective tissue growth factor.
Wei-Wei, YU ; Qin, XIA ; Yan, WU ; Qiao-Yun, BU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(5):652-6
Long-term treatment with an agonist of peroxisome proliferator-activated receptor (PPAR)-γ is associated with bone fractures in the clinical practice. However, the mechanisms underlying the fractures are not fully understood. This study was aimed to examine the effect of rosiglitazone (an agonist of PPAR-γ) of different doses on the proliferation, differentiation, and transforming growth factor beta 1 (TGF-β1)-induced expression of connective tissue growth factor (CTGF) in primary rat osteoblasts in vitro. Osteoblasts were isolated from newly born SD rats and treated with different doses of rosiglitazone (0-20 μmol/L). The proliferation and differentiation of osteoblasts were measured by MTT assay and NPP assay, respectively. The expression of CTGF was determined by RT-PCR and Western blotting. The results showed that most isolated osteoblasts displayed strong alkaline phosphatase (ALP) activity and treatment with different doses of rosiglitazone did not affect their proliferation, but significantly inhibited the differentiation of osteoblasts in a dose-dependent manner. Moreover, treatment with different doses of rosiglitazone significantly reduced the TGF-β1-induced CTGF mRNA transcription and protein expression in a dose-dependent manner in rat osteoblasts. It was concluded that the activation of PPAR-γ may inhibit the differentiation of osteoblasts by reducing the TGF-β1-induced CTGF expression in vitro.