1.Ultra-structural changes of the lumbar spinal motor neurons after sciatic nerve injury induced by high-speed steel ball
Zhiyong YIN ; Ping ZHOU ; Hong HUANG
Chinese Journal of Tissue Engineering Research 2005;9(14):-
BACKGROUND: What kind of traumatic changes could be observed in remote lumbar spinal cord after high-speed steel ball induced sciatic nerve injury?OBJECTIVE: To study the ultra structural changes of lumbar spinal cord motor neurons after the sciatic nerve injury due to high-speed steel ball.DESIGN: Retrospective randomized controlled study with experimental animals as subjects.SETTING: The field operation research institute of a military medical university college.MATERIALS: This study was carried out at the Ballistic Laboratory of Research Institute for Surgery from May 2001 to December 2002. Sixty-five SPF flap-eared rabbits of either gender weighing from 2.5 to 3.0 kg were provided by the Experimental Animal Center of Daping Hospital Affiliated to the Third Military Medical Univeristy of Chinese PLA. All the rabbits were divided randomly into normal control group( n = 5), firearms injury group (n = 30) and cutting injury group( n = 30) . The two latter groups were subdivided into one-day, three-day, one-week, two-week, 4-week and twelve-week groups with 5 rabbits in each group.METHODS: The rabbits in the firearms injury group were injured at the midpoint of lateral posterior body surface projection of the right sciatic nerve with 0. 38 g steel ball containing 0.65 g detonators. The right sciatic nerve was cut at the same injury level in the cutting injury group.MAIN OUTCOME MEASURES: The following pathologic changes of the lumbar spinal cord were observed with optical and electronic microscope at posttraumatic day 1, day 3 and week 1, week 2, week 4 and week 12 respectively.RESULTS: Micro-hemorrhage, neuron swelling and vacuoles were observed in the firearms injury group and the number of motor neurons decreased significantly, but these changes were unobservable in the cutting injury group.CONCLUSION: Compared with the cutting group, the injury to the spinal cord was severer in the firearms group and the survival neurons were less.
2.DELETION ANALYSIS OF DNA FRAGMENT RM07 FROM HALOBACTERIUM HALOBIUM
Yin WANG ; Yu-Ping HUANG ; Zhen-Hong DUAN ; Ping SHEN ;
Microbiology 1992;0(01):-
The DNA fragment RM07 was isolated from halophilic archaea Halobacterium halobium, which can function as promoter not only in halophilic archaea, but also in Escherichia coli as eubacterial promoter. Sequencing analysis indicated that it possessed the typical consensus sequences (-35 and -10) of bacterial gene promoter, which was confirmed by further deletion analysis: With its -35 sequence deleted and -10 sequence left,DNA fragment RM07a nearly cannot initiate transcription;With its both -35 and -10 sequences,RM07b DNA fragment could be active as promoter at a level even higher than RM07. Our research also showed that the promoter function of RM07 fragment in Escherichia coli was under the control of environmental factors,especially its positive correlation with the increasing concentration of sodium chloride. Therefore, RM07 DNA fragment may be potential1 novel promoter source for constructing double-function vectors. It also has special significance in elucidating the issues of the fusing characteristics of archaea and lateral gene transfer between archaea and bacteria.
3.Diagnostic value of endoscopic ultrasonography for ulcerative colitis
Ping YIN ; Zhenhua YANG ; Aoshuang HUANG ; Fei SHI ; Yan MAO
Chinese Journal of Digestive Endoscopy 2012;23(1):6-10
Objective To study the consistency and complementarity of endoscopic ultrasonography (EUS),white light endoscopy (WLE) and magnifying endoscopy (ME) in diagnosis of ulcerative colitis (UC).Methods We collected 125 cases of UC patients diagnosed by WLE and EUS (including 51 cases of WLE + ME + EUS).According to UC mucosal morphology under WLE and crypt openings under ME,we divided all the cases into several groups and analyzed intestinal wall thickness (TWT) under EUS in each group.Results According to the results of UC inflammation degree under WLE,all patients were divided into four groups: 16 severe cases,46 moderate cases,44 mild,and 19 remission stage.TWT results were (5.903 ± 1.551 ) mm,(4.673 ± 1.235 ) mm,(3.756 ± 1.322 )mm and ( 3.464 ± 0.970) mm,respectively.Differences were significant between any two groups ( P < 0.05 ),except for that between mild and remission groups.According to the results of UC inflammation degree under ME,all patients were divided into six groups: 9 cases of villous-like structure,9 cases of typical coral reef-like structure,8 severe coral reef-like structure,13 regular crypt opening,6 epithelial minimal defect and 6 small yellow spot (SYS).TWT results were (5.701 ±0.941 )mm,(5.518 ±0.581 )mm,(5.181 ±0.751 )mm,(3.763 ±0.659) mm,(3.587 ±0.461 )mm and (2.505 ± 0.330 )mm,respectively.Differences were significant between any two groups ( P < 0.05 ) except for those between epithelial minimal defect and regular crypt opening,typical coral reeflike structure,villous-like and severe coral reef-like structure.EUS results showed SYS (6/6) and regular crypt opening ( 10/13 ) were mostly located in mucosa,while lesions of severe coral reef-like structure (8/8) invaded the muscularis propria.Conclusion EUS shows high consistency with WLE and ME in diagnosis of UC inflammation degree and invasive depth.It could assist and even substitute ME for evaluation.
4.Application of vascularized anterolateral thigh perforator flap harvested with free-style approach in oral and maxillofacial reconstruction
Jie CHEN ; Canhua JIANG ; Ping YIN ; Long YANG ; Xiaoshan WU ; Long HUANG ; Xinchun JIAN
Chinese Journal of Microsurgery 2015;38(1):20-24
Objective To assess the harvest method and application value of free-style anterolateral thigh perforator flap (ALTPF) in oral-maxillofacial reconstruction.Methods Fifty-three patients who suffered from oral and maxillofacial tumor underwent simultaneous reconstruction using free-style ALTPFs after radical resection from August,2013 to May,2014.Perforators of the ALTPF region were marked through hand-held Doppler probe preoperatively.Flaps were designed centered on perforators according to the defect size of the accepting site.Incisions were first made at the inner border of the designed flap.Perforators were exposed above the fascia lata femoris and then dissected retrogradely through the vastus lateralis muscle to harvest a vascular pedicle with desired caliber and length.Flap thinning was applied under microscope in some cases to compromise the need of the accepting site.Results All 53 flaps survived after transplantation while skin exfoliation occurred in 5 cases due to flap thinning.Four cases sustained partial necrosis and was cured by trimming and dressing changes.Five to 14 months' post-operative followup showed satisfactory accepting-site morphology with good speech function and swallowing recovery.All donor sites were closed primarily without skin-grafting,leaving no donor-site complications including incision disruption,scar hyperplasia and muscle strength degeneration of the lower limb.Conclusion Multiple perforators have been accu rately located preoperatively in free-style harvest approach of ALTPFs,thus optimal effects can be reached with decreased donor-site morbidity and improved aesthetic outcome to the uttermost,which accords with the refined,personalized and minimal invasive development concept of modem reconstructive surgery.
5.Survival of bone marrow mesenchymal stem cells after transplantation into the rat infarcted myocardium
Chuwei LIN ; Shenghua ZHOU ; Haiying DAI ; Ping DENG ; Hongguang HUANG ; Zhilan YIN ; Xiansong GUAN
Chinese Journal of Tissue Engineering Research 2014;(41):6628-6632
BACKGROUND:The preliminary findings confirm that bone marrow mesenchymal stem celltransplantation is safe and effective in the treatment of acute myocardial infarction, but its exact mechanism is unclear. There are few studies addressing the survival status of transplanted stem cells and its acting timing.
OBJECTIVE:To study the survival of rat bone marrow mesenchymal stem cells transplanted into the infracted myocardium. METHODS:Bone marrow mesenchymal stem cells were cultured using density gradient centrifugation. Eighty rat models of myocardial infarction were prepared. Bone marrow mesenchymal stem cells were injected via a microsyringe at four sites around the infarcted region at 14 days after modeling. Then, 70 rats with living cells were selected for detecting the survival of bone marrow mesenchymal stem cells at days 3, 5, 7, 10, 20, 28 after transplantation. RESULTS AND CONCLUSION:Under ×400 visual field, the number of Brdu-positive bone marrow mesenchymal stem cells was (36±12) at 3 days posttranplantation, (33±13) at 5 days, (28±9) at 7 days, (15±5) at 10 days, (5±3) at 14 days, 0 at 20 days, and 0 at 28 days, showing a overal downward trend after transplantation. The number of bone marrow mesenchymal stem cells was negatively correlated with transplant days (P<0.01, r=-0.47). The number of cells decreased most significantly within 1 week after transplantation, and then decreased to 0 at 20 days. These findings indicate that transplanted bone marrow mesenchymal stem cells in the myocardium cannot survive for a long term and also cannot be transformed into myocardial tissue.
6.Study on protective effect of astragaloside Ⅳ in septic mice
Baohong YUAN ; Ping HUANG ; Xinmeng DENG ; Keying WANG ; Liangcheng DAI ; Hui YIN
Chinese Pharmacological Bulletin 2017;33(10):1452-1456
Aim To study the effect of astragaloside (AS-Ⅳ) in CLP-induced septic mice.Methods C57BL/6 mice were randomly divided into the sham group, CLP group and CLP+ AS-Ⅳ group.Two days before operation, AS-Ⅳ (10 mg·kg-1) solution was intragastrically administered into CLP +AS-Ⅳ group, and the other groups were treated with normal saline.A sepsis model was established by cecal ligation and puncture (CLP).Blood, peritoneal fluid and tissue organs were collected at 6 h and 24 h.Neutrophils of blood were purified by Percoll density gradient.Transwell was used to detect the chemotaxis function of neutrophils.The killing activity of neutrophils was detected by coculture with E.coli.Results The survival rate of AS-Ⅳ-pretreated septic mice significantly increased.The number of neutrophils in peritoneal fluid was enhanced markedly.The number of bacteria in the peritoneal fluid, blood and tissue organs such as liver, lung and kidney significantly decreased after AS-Ⅳ pretreatment.The chemotaxis and killing activity of neutrophils increased significantly in AS-Ⅳ-treated mice (P<0.05).Conclusion Astragaloside displays an immunoprotective effect in CLP-induced septic mice, which is related to the upregulation of CXCR2 expression on neutrophils and the increase of neutrophil antibacterial activity.
7.Package and identification of replication deficient recombinant adenovirus expression vector of channelrhodopsin-2
Jun-ping, YAO ; Wen-sheng, HOU ; Hao, WANG ; Chuan-huang, WENG ; Zheng-qin, YIN
Chinese Journal of Experimental Ophthalmology 2012;30(8):681-685
Background Channelrhodopsin-2 (ChR2)is a cation channel isolated from the eyespot of Chlamydomonas algae and has been used to control neuron activity.The light stimulation is a more precise fashion whether space or time than that of electrical,magnetic and ultrasound stimulation. Objective This study was to construct a replication deficient recombinant adenovirus cxpression vector of ChR2 and to determine its function.Methods Human embryo kidney 293 (HEK293) cell line was cultured and passaged in DF12 medium containing 10% fetal bovine serum(FBS).The ChR2 gene was cloned at the downstream of cytomegalovirus(CMV)promoter of the adenoviral shuttle plasmid pSB291 in sense direction,and the resultant recombinant plasmid pSB291-hChR2- GFP was transfected into HEK293 cell together with plasmid pBHG lox ( deltaE1,3 ) containing adenoviral genome,then small amounts replication deficient recombinant adenovirus expression vector of ChR2 (Ad-ChR2) was obtained.Through amplification gradient centrifugation and dialysis,pure Ad-ChR2 was obtained.Visual cortex cells derived from 4 1-day-old clean Long Evans rats were primary cultured with serum-free culture media and infected by AdChR2.When expressing green fluorescencc,those cells received the stimulated of blue light with 460 nm.Patch clamp technique was applied to record an action potential. Results After purification and concentration,the titer of AdhCHR2 reached 7.9×1010 PFU/ml.Twenty-four hours after transfect of Ad-ChR2,HEK293 cell membrane showed the green fluorescence for the recombinant plasmid with green fluorescence protein under the inversed fluorescence microscope.The HEK293 cells change their shape from flat to round 13 days after transfected.The primary cultured visual cortex cells exhibited the green fluorescence 3-5 days after infected by Ad-ChR2.The action potentials evoked by blue light stimulation were recorded with patch clamp on those cells expressing green fluorescence. Conclusions Ad-ChR2 expressing vector is constructed successfully in this study.It is verified that Ad-ChR2 expressing vector can infect visual cortex cells with visual function.This result is very important for visual plasticity study.
8.Pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation on recurrent pterygium
Ting, LI ; Shu-Xian, CHEN ; Xu-Guang, XIA ; Zheng, YIN ; Mu-Zhen, HUANG ; Ping-Ying, GUO
International Eye Science 2014;(9):1715-1716
To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.
9.Perinatal outcomes of 45 pregnant women with pulmonary hypertension complicating congenital heart disease
Yuan MENG ; Yin-Ping HUANG ; Xiao-Li LIU ; Yan-Jun HUANG ; Jie ZHOU ; Chun-Qiu XIONG ; Li-Qun WU ; Bo WANG ; Cai-Ping CAI
Chinese Journal of Obstetrics and Gynecology 2000;0(10):-
Objective To explore the perinatal outcomes of women with pulmonary hypertension complicating congenital heart disease(CHD).Methods Clinical data of 45 cases of pregnant women with pulmonary hypertension complicating CHD from Apr 1995 to May 2007 were analyzed and they were divided into three groups:29 cases of slight group[pulmonary hypertension of 30 mm Hg(1 mm Hg=0.133 kPa) to 49 mm Hg],8 cases of moderate group(pulmonary hypertension of 50 mm Hg to 79 mm Hg)and 8 cases of severe group(pulmonary hypertension equal to or higher than 80 mm Hg).The types of CHD,cardiac functional status(New York heart association,NYHA),gestational weeks of pregnancy termination,mode of delivery,pregnancy after CHD operation and outcomes of infants were compared between the groups. Results(1)The highest incidence of CHD were atrial septal defect and ventricular septal defect(58%, 26/45).The rate of pregnant women after CHD operation was 29%(13/45),they were mainly in slight group and their NYHA class were in Ⅰ-Ⅱ.(2)The occurrence rate of NYHA class Ⅲ-Ⅳ was 7/8 in severe group.The rate of NYHA class Ⅰ-Ⅱ as 6/8 in moderate group.The rate of NYHA class Ⅰ- was 97%(28 /29)in slight group.(3)The rate of term delivery was 93%(27/29),preterm labor 3% (1/29),abortion 3%(1/29),and the birth weight was(3153?399)g on average in slight group.The rate of term delivery was 5/8,preterm labor occurred in 3 cases in moderate group.The rate of term delivery was 5/8,preterm labor occurred in 2 cases,and iatrogenic abortion in 1 case in severe group.The average birth weight between slight group and moderate or severe group had a significant difference.(4)Caesarean section rate was 78 %(35/45)among all patients.The rate of cesarean section delivery was 76%(22/29)in slight group,6/8 in moderate group,and 7/8 in severe group.(5)The rate of pregnant women who had portent heart failure or heart failure was 24%(11/45),overall maternal mortality was 4%(2/45).Conclusions The higher the pulmonary hypertension,the worse the outcome of the mother and fetus;The pregnant women with good heart function after cardiac operation would have a good perinatal outcome.Cesarean section is more suitable for those women.
10.Influence of mesenchymal stem cells on proliferation of HL-60.
Wen-Jie YIN ; Xiao-Peng LIU ; You-Zhang HUANG ; Ping-Di YANG
Journal of Experimental Hematology 2012;20(1):62-65
The objective of this study was to explore the effect of mesenchymal stem cells (MSC) on HL-60 proliferation and its molecular mechanism. HL-60 cells co-cultivated with MSC were used as experiment group, and the cells cultivated solely were taken as control group. HL-60 cells in the two groups were counted at different time. The time-quantity curve was drawn. The cell cycle and apoptosis ratio of HL-60 cells were compared between the two groups and expressions of Survivin and Bcl-2 protein were detected by flow cytometry. The results showed that HL-60 cells cultivated with MSC were obviously inhibited, especially on day 5 and 7 (P < 0.01). HL-60 cells were distributed on the phase of G(0)/G(1) [control group (47.0 ± 9.0)% vs experiment group (70.0 ± 16.0)%, P = 0.003], and apoptotic peak appeared. Both of Survivin and Bcl-2 protein expressions in HL-60 cells decreased [Bcl-2 protein in control group (63.0 ± 9.1)% vs experiment group (50.0 ± 14.1)%, P = 0.045; Survivin in control group (94.0 ± 9.3)% vs experiment group (77.0 ± 11.8)%, P = 0.006]. It is concluded that the MSC can inhibit HL-60 cell proliferation, and promote HL-60 cell apoptosis.
Adult
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Apoptosis
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Bone Marrow Cells
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cytology
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Cell Proliferation
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Coculture Techniques
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Flow Cytometry
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HL-60 Cells
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Humans
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Inhibitor of Apoptosis Proteins
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metabolism
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Mesenchymal Stromal Cells
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cytology
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Middle Aged
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Proto-Oncogene Proteins c-bcl-2
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metabolism