1.Role of C-terminal Src kinase in angiotensin Ⅱ-induced cytoskeletal rearrangement in glomerular podocytes
Lu ZHANG ; Zhilong REN ; Qian YANG ; Yiqiong MA ; Guohua DING
Chinese Journal of Nephrology 2015;31(11):842-847
Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of C-terminal Src kinase (Csk) in Ang Ⅱ-infused rat model and cultured podocytes,and to explore the role of Csk in Ang Ⅱ-induced cytoskeletal rearrangement of podocytes.Methods Twenty-four Wista rats were randomly subjected to normal saline infusion,or Ang Ⅱ infusion at 400 ng · kg1 · min-1 (via subcutaneous osmotic minipumps) for 2 or 4 weeks.Renal histomorphology was evaluated through electron microscopy.The expression of glomerular Csk was analyzed by immunofluorescence and Western blotting.In vitro,conditionally immortalized mouse podocytes were cultured and treated with Ang Ⅱ doses ranging from 10-9 mol/L to 10-5 mol/L and for different hours.The expression of podocytes Csk was assessed by Western blotting.After transfection to podocytes with Csk siRNA,FITC-conjugated phalloidin was used to stain F-actin,to investigate the role of Csk in Ang Ⅱ-induced or cytochalasin D-induced cytoskeletal rearrangement.Results (1) Examination of Ang Ⅱ infusion rats glomerular and podocyte ultrastructure by electron microscopy revealed foot process effacement and fusion; (2) In Ang Ⅱ infusion rats,the expression of glomerular Csk was increased (P < 0.05); (3) In vitro,Ang Ⅱ-stimuli up-regulated the expression of Csk (P < 0.05),and the effects of Ang Ⅱ were on dose-dependent and time-dependent manner; (4) Ang Ⅱ-induced disruption of F-actin was alleviated by Csk siRNA transfection in cultured podocytes; furthermore,cytochalasin D depolymerized the F-actin cytoskeleton,while Csk siRNA stabilized the actin filaments.Conclusion The enhanced expression of Csk may be involved in Ang II-induced podocytes cytoskeletal rearrangement and foot process fusion.
2.The cognitive development of conflict monitoring of school-age children: a behavior and ERP study
Jing CAI ; Yanling REN ; Suhong WANG ; Yili ZHANG ; Jian CAO ; Zhilong YANG ; Ling MA ; Xuan DONG
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(1):22-24
Objective The present study aimed to trace the behavior results and event-related potential (ERP) of conflict monitoring from 7 ~ 12 years old to explore the development features of the conflict monitoring.Methods In six groups of 144 children aged from 7 to 12,behavior and non-target N2 amplitude were analyzed in continuous performance test (CPT) task. Results 1. Behavior results:the reaction time of target stimulus decreased ( ( 533.33 ± 66.65 ) ms, (523.91 ± 92.96 ) ms, (468.37 ± 64. 13 ) ms, ( 46 1.48 ± 98.31 ) ms, (457.57 ±84.05 ) ms, (405.02 ± 67.90) ms) and the hitting number increased ( ( 34.87 ± 4.84 ), ( 37.64 ± 3.54 ), ( 37.95± 2.92 ), (38.67 ± 1.23 ), (39.31 ± 1.08 ), ( 39.45 ± 1.00 ) ) as age increased, and the difference was statistically significant ( P<0.01 ). 2. ERP: ①The non-target N2 amplitude was significantly higher than the target,and the difference was statistically significant (F= 98.57, P< 0.01 ). ②The amplitude of non-target N2 amplitude decreased with age, and the difference was statistically significant (F= 5.54, P< 0. 01 ). Conclusion The non-target N2 was closely related to the monitoring conflict, and the behavior and ERP results in this study showed the development trend. 8 ~ l0 and 12 years old are the critical development period of information processing speed, attention and conflict monitoring function for children.
3.The conflict monitoring of children with attention deficit hyperactivity disorder:an event-related potentials study
Jian GAO ; Suhong WANG ; Zhilong YANG ; Yili ZHANG ; Jing CAI ; Yanling REN ; Ling MA ; Xuan DONG
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(11):975-977
Objective To compare the event-related potential(ERP) difference waveforms between attention deficit hyperactivity disorder (ADHD) children and normal children in the conflict task,and to investigate the neural mechanism of conflict monitoring of ADHD. Methods ERPs elicited by performing the spatial SimonStroop task were recorded in 32 children with ADHD and 32 normal children. The differences of ERP and behavior dren with ADHD(0.80±0. 11 ,0.87 ±0.08) were less than normal children (0.88 ±0.07,0.93 ±0. 04)in Sireaction time of ADHD group( (773.03 ±99.34)ms) in the Siin were longer than the control group( (737.21 ±81.40)ms) ,the differences had statistic significance (P<0. 05). 2. ERP results:the amplitude of N2 of the children with ADHD in the four conditions were smaller than the control group, the difference in Siin had statistic sigof prefrontal and anterior cingulate cortex of ADHD were weaker than the control group. Conclusions The results suggest that, due to the inadequate allocation of attention resources of the children with ADHD, they have conflict monitoring function defects compare with normal children. The function defects of conflict information processing related brain areas may be the main neural mechanism. The result of the behavior and ERP can become the reference indicator for the early diagnosis and treatment of ADHD.
4.Brain imaging studies in children with attention-deficit hyperactivity disorder revealed by resting-state fMRI fALFF analysis
Zhilong YANG ; Suhong WANG ; Jian CAO ; Yanling REN ; Jing CAI ; Yili ZHANG ; Ling MA ; Xuan DONG
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(10):894-896
Objective To investigate pathological mechanism of dysfunction of executive control in children with attention-deficit hyperactivity disorder (ADHD) ,using a new resting-state functional magnetic resonance imaging(fMRI) index, fractional amplitude of low frequency fluctuation (fALFF). Methods Thirty children of school age with ADHD and thirty gender, age and education-matched healthy controls were experienced restingstate fMRI scans. fALFF approach was used to analyze blood oxygen level-dependent fMRI(BOLD-fMRI) data in resting state. Results Compared with the normal controls,the ADHD showed decreased fALFF in the bilateral superior frontal gyrus( BA10, t=5. 40,4.11 ,Z =4.81,3.82),bilateral middle frontal gyrus( BA9,BA46, t=5.72,5.37, Z = 5.04,3.08 ), right medial frontal gyrus ( BA 10, t = 5.53, Z = 4.90 ) and left precuneus ( t = 3.81, Z =3.57). And the ADHD showed increased fALFF in the bilateral limbic lobe (BA36, t = 4.37,5.20, Z = 4.02,4.66 ), right brainstem ( t = 4.77, Z = 4.34) and bilateral cerebellum ( t = 4.65,4.83, Z = 4.24,4.38 ). Conclusion The results suggest that lower activition of frontal may be the core deficit of executive control with ADHD in resting state,and high activation in several brain regions may be related to compensatory effect.
5.Angiotensin Ⅱ induces podocyte c-Abl expression change
Xinghua CHEN ; Zhilong REN ; Tean MA ; Dongqing ZHA ; Cheng CHEN ; Guohua DING
Chinese Journal of Nephrology 2012;28(7):548-552
ObjectiveTo evaluate the effects of angiotensin Ⅱ (Ang Ⅱ )infusion on renal c-Abl expression in vivo,and on podocyte c-Abl expression change in cultured mouse podocytes.Methods Twenty four male Sprague-Dawley rats (Group C,D,E and F) were assigned to receive Ang Ⅱ(400 ng· kg-1 min-1) by osmotic minipump and of which 12 rats (Group D and F) were assigned to receive telmisartan (3 mg·kg-1·d-1),six rats received normal saline(Group B),and six rats were used as normal control(Group A).Animals were sacrificed at day 14 (Group C and D),day 28 (Group E and F) respectively.Conditionally immortalized mouse podocytes were used in vitro.Podocytes were studied 2 weeks after thermoswitching from 33℃ to 37℃.Cells were fetal bovine serum(FBS) starved for at least 12 hours prior to stimulation.The cultured podocytes were treated withAngⅡdosesranging from10 -9 mol/L to10 -6 mol/L andfor differenthours.Expression of renal and podocytes c-Abl was examined by immunofluorescence staining,real-time PCR and Western blotting.Results(1) Distribution of c-Abl expression was mainly in the cytoplasm and nuclear of the podocytes in vivo and in vitro. (2) Expressions of c-Abl mRNA and protein wereincreasedinAng Ⅱ-infusedratpodocytesandAng Ⅱ-inducedculturedmouse podocytes(P<0.05),and the effects of Ang Ⅱ were dose-dependent and time-dependent in vitro.Conclusion There are c-Abl mRNA and protein expression in podocytes,and c-Abl may play a critical role in the pathogenesis of Ang Ⅱ -induced podocyte injury.
6.Experimental study of skin wound healing with epidermal stem cells from human hypertrophic scar in nude mice.
Shuping ZHOU ; Zhilong HUANG ; Jiguang MA ; Jingjing XU ; Jinglong CAI ; Xianlei ZONG ; Le DU
Chinese Journal of Plastic Surgery 2014;30(4):289-293
OBJECTIVETo investigate the effect of epidermal stem cells from human hypertrophic scar (HS-ESCs) on the skin wound healing in nude mice.
METHODS40 mice were randomly divided into two groups as experimental group (n = 20) and control group (n = 20). Wounds, 1 cm in diameters, were made on every mouse back. The wounds were treated with HS-ESCs and erythromycin ointment in experimental group, or only with erythromycin ointment in control group. The wound healing was observed during the following 14 days. The expression of collagen-I, collagen-III, epidermal growth factor (EGF), fibroblast growth factor (FGF2) , transforming growth factor (TGFbeta1, and TGFbeta2) were studied.
RESULTSThe wound healing time in the experimental group was (20.8 +/- 0.84) d, which was (25.6 +/- 0.89) d in the control group. HE staining revealed that the extent of vascularization in the experimental group was 11.60 +/- 0.55, while it was 8.04 +/- 0.33 in the control group. Immunochemistry analysis showed the expression of collagen-I, collagen-III, EGF, FGF2, TGFbeta1, and TGFbeta2 in the experimental group were significantly higher, compared with those in control group (P < 0.05).
CONCLUSIONHS-ESCs may promote wound healing through enhancement of the vascularization of the wound tissue and the expression of growth factors.
Animals ; Cicatrix, Hypertrophic ; pathology ; Epidermis ; cytology ; Female ; Humans ; Male ; Mice ; Mice, Nude ; Skin ; injuries ; Stem Cell Transplantation ; Stem Cells ; Wound Healing
7.Purify a Modifier Protein of Glyceraldehyde-3-Phosphate Dehydrogenase
Qing REN ; Haidong YAN ; Lan WU ; Zhilong LI ; Jianfei MA ; Lining WONG ; Xijing ZHOU
Journal of China Medical University 2001;30(1):31-32,40,43
Objective: Our aim was to purity the modifier protein of glyceraldehyde-3-phosphate dehydrogenase (G3PD) from African green monkey Vero-E6 line. Methods:Exposure of Vero-E6 cells to medium with a reduced K concentration (3.2 mmol/L) stimulated the growth and activation of G3PD. The increase of enzyme activity was mediated by a cytosolic modifier protein that was purified using affinity and anion-exchange high-performance liquid chromatograph. Results:The apparent molecular mass of the protein was 62 kDa. Western blotting and quantiative enzyme-linked immunosorbent assay showed that the amount of modifier protein increased progressively for 2 hours in cells exposed to low-K+ medium, and then returned to the control value, a kinetic profile similar to that the modifier protein is a constituent of renal epithelial cells and accummulated transiently in the low-K+ mitogenic signal. Conclusion: We obtained a modifer protein from monkey kidney epithelial cells (Vero-E6). It could activate G3PD and cell growth.
8. Study of epidemic area on Tsutsugamushi disease in Taizhou from 2013 to 2014
Yilin HE ; Haiyu YANG ; Chunxia YU ; Xiang ZHANG ; Qianhua YI ; Zhilong MA ; Jie CHA ; Xiaobing XU ; Qi ZHANG ; Wenjun DAI ; Weijuan QIAN ; Jun YIN ; Saijin ZHU ; Zhen XU
Chinese Journal of Preventive Medicine 2017;51(3):252-256
Objective:
To study the epidemiological characteristics of tsutsugamushi disease, and to confirm the existence of the disease's epidemic foci in Taizhou.
Methods:
From 2013 to 2014, Dongxing town hospital and Xingqiao town hospital were selected as specimen collection sites in Jingjiang city. Blood samples (5 ml) were collected from 40 patients with acute tsutsugamushi disease. A total of 59 rodents were captured with cage night method in the survey sites at 5, 7, 9, 10, and 11 months in 2013, from which, the spleen, liver, and kidney specimens were selected. Chigger mites were captured by small blackboard method and from the ears of the captured rodents. A total of 226 small blackboards were laid, 27 mites were captured, and the samples were grounded into suspension. Nested-polymerase chain reaction and cell and tissue culture techniques were used to test the specimen from the probable patients, host animals and chigger mites.
Results:
Among the 40 acute tsutsugamushi disease blood samples, 29 were found to meet the test requirements, 17 were positive for orientia tsutsugamushi nucleic acid with 59% of the positive rate, and 1 stran orientia tsutsugamushi was isolated. 59 rats were captured and the density of mice was 5.5%. Among them, there were 26 Mus musculus (2.4%), 18 Rattus flavipectus (1.7%) and 15 Smelly shrew (density 1.4%). 1 Smelly shrew was tested positive for orientia tsutsugamushi nucleic acid, and the negative results were found in the other rodent specimens. 27 Chigge mites were collected by small blackboard method and the density of mites was 0.12 for each blackboard, among which 3 larvae and 24 nymphs were found. 33 Chigger mites were collected from the ears of 3 Smelly shrew, and the density of the mite was 11 per mouse. All the captured Chigger mites were identified as Leptotrombidium scutellare and 1 group of specimens of Chigger mites from the external environment were positive for orientia tsutsugamushi nucleic acid.
Conclusion
There was a high density of mice in the epidemic area from May to November and the species of the chigger mites were Chigger mites in Taizhou. The nucleic acid of the oriental tsutsugamushi was detected in the patients with acute scrub typhus, rodents and vectors. According to the above-mentioned results, it was considered that the scrub typhus epidemic area of Taizhou city has the natural foci of scrub typhus.
9.Application of posterior tibial artery perforator flap with great saphenous vein in treatment of forearm degloving injury and reconstruction of venous circulation of dorsal hand
Shengjun YU ; Zhenjie MA ; Shengqiang FU ; Xiao LI ; Zhilong WANG ; Juntao XU ; Ying LI
Chinese Journal of Microsurgery 2022;45(4):372-376
Objective:To explore the clinical application of the posterior tibial artery perforator flap with the great saphenous vein (GSV) in the treatment of severe degloving injury of the forearm.Methods:From June 2015 to October 2020, 5 patinets (4 males and 1 female, aged 20-46 years old, mean age 37 years old) were treated in the Department of Hand Surgery of Yantaishan Hospital in Yantai. Aposterior tibial artery perforator flap with GSV was used to repair the partial wound of the injured forearm, together with the establishment of venous circulation of dorsal hand for all 5 patients. All the patients suffered from severe forearm degloving injury. Of which, 3 accompanied with ulna radius fracture, 2 with ulnar and radial artery injury and 2 with blood supply insufficiency in the injured fingers. The sizes of soft-tissue defect were 26 cm×18 cm-32 cm×25 cm. The sizes of the posterior tibial artery perforator flap with GSV ranged from 12 cm×5 cm to 33 cm×6 cm. The anastomoses were performed on the perforating artery and the radial artery. The GSV was anastomosed with cephalic vein with bridging anastomosis to re-establish the venous circulation of dorsal hand. Combined anterolateral thigh flaps (ALTF) were used to repair the rest wound of injured limbs in 4 patients, and the combined pedicled abdominal flap was used in 1 patient. The donor site of calf flap was sutured directly, and the skin of upper limb was thinned into medium thick to cover the thigh flap donor site, which was packed with pressure dressing. Regular follow-up reviews were carried out by outpatient clinic, telephone, WeChat APP or home-visit after the operation to observe the survival of flaps, the swelling of the distal end of injured limb, functional recovery and healing at the donor site. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Results:All the grafted flaps in 5 patients survived. Necrosis at epidermal edge occurred in 1 calf flap, and achieved secondary healing after dress changing for 4 weeks. There was neither arterial nor venous crisis in all flaps. The oedema of the hand disappeared within 4-8 weeks, with clear dermatoglyph. There was no obvious enlarged circumference of thumb and fingers. The follow-up was carried out for 8-20 months, with 11 months in average. The flaps were elastic in good colour, and full texture without pigmentation. The donor site of the calf flap showed a linear scar. No swollen was seen in the hands of the injured forearm and the feet of donor lower legs. The range of motion of phalangeal joints was good. Recovery of finger sensation achieved at S 4. The recovery of the sensation of posterior tibial artery perforator flap reached S 3 in 2 patients and S 2 in 3 patients. Assessment of the Upper Limb Function using the Standard Issued by the Hand Surgery Society of Chinese Medical Association rated excellent in 3 patients and good in 2 patients. Conclusion:The posterior tibial artery perforator flap with GSV has a reliable blood supply with a small damage to the donor site. This flap is ideal for repair of severe degloving injury of forearm and meanwhile to achieve the re-establishment of venous circulation in dorsal hand.
10.Application of anterior lateral malleolar venous network graft in repair of defect of superficial palmar arch: Report of 8 cases
Shengjun YU ; Zhenjie MA ; Xiao LI ; Shengqiang FU ; Zhilong WANG ; Juntao XU ; Ying LI
Chinese Journal of Microsurgery 2022;45(6):629-633
Objective:The surgical method and clinical efficacy was studied on the anterior lateral malleolar venous network grafting in repair of the defects of superficial palmar arch.Methods:From February 2015 to July 2021, 8 cases (6 males and 2 females; aged 32-44 years old, mean age at 39 years old) were treated in the Department of Hand Surgery of Yantaishan Hospital. Anterior lateral malleolar venous network was grafted to repair the defects of superficial palmar arch of the patients. The patients had different degrees of tendon damage, and injuries of nerve, deep palmar arch, common palmar digital artery or proper palmar digital artery. The length of the vessel defect was 6.0-10.0 cm. Blood supply insufficiency occurred in 1st-5th digits in 1 case, 2nd-4th fingers in 5 cases, and 2nd-5th fingers in 2 cases. The anterior lateral malleolar venous network was transferred to repair 3 broken ends of vessels in 5 cases, 4 broken ends in 1 case and 5 broken ends in 2 cases. The wound of hand was sutured directly in 2 cases. The emergency surgical repair of the wound of hand by transfer of ALTPF was performed in 2 patients, and elective ALTPF transfer in 3 cases. Forearm dorsi-ulnar flap transfer was performed in 1 case. The follow-up reviews were carried out via the outpatient clinic visit, telephone, WeChat interviews or home visits. The blood supply of the distal digit, skin sensation, TPD, the range of motion of the phalangeal joints were observed.Results:The results showed that all the digits and the grafted flaps survived. The follow-up was carried out for 10-53 months, with an average of 17.9 months. The functions of digit extension and flexion, finger opposition and thumb opposition were restored in 5 cases, and the patients returned back to former jobs. The finger flexion and thumb opposition were limited in 3 cases, and these cases participated in light physical work. The proprioception of the digits were recovered, and the TPD was 5-7 mm. Functional recovery was evaluated according to the Evaluation Standard of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association and rated as excellent in 5 cases and good in 3 cases. The donor sites of the foot showed a linear scar. There was no effect on the function and the shape of the donor site.Conclusion:The anterior lateral malleolar venous network grafting is an ideal method in repair of the defects of superficial palmar arch.