1.Advances in clinical application of laparoscopic totally extraperitoneal herniorraphy
Yujian ZENG ; Tong ZHANG ; Chengmin SHI ; Zhizhong WU ; Yanqiang SHI ; Huayou LUO
Chinese Journal of Digestive Surgery 2022;21(9):1197-1200
Inguinal hernia is a common surgical disease, and most patients need surgical treatment. In recent years, minimally invasive surgery based on laparoscopy has been popularized in hernia surgery. With the release of clinical guidelines, the progress of instruments and materials, the update of treatment concepts and anatomical knowledge, laparoscopic inguinal hernia repair, especially laparoscopic total extraperitoneal hernia repair (TEP), is developing towards a more accurate and minimally invasive direction. Based on literatures in recent years and combined with clinical practice, the authors explore the advances in clinical application of laparoscopic TEP.
2.High-throughput "read-on-ski" automated imaging and label-free detection system for toxicity screening of compounds using personalised human kidney organoids.
Qizheng WANG ; Jun LU ; Ke FAN ; Yiwei XU ; Yucui XIONG ; Zhiyong SUN ; Man ZHAI ; Zhizhong ZHANG ; Sheng ZHANG ; Yan SONG ; Jianzhong LUO ; Mingliang YOU ; Meijin GUO ; Xiao ZHANG
Journal of Zhejiang University. Science. B 2022;23(7):564-577
Organoid models are used to study kidney physiology, such as the assessment of nephrotoxicity and underlying disease processes. Personalized human pluripotent stem cell-derived kidney organoids are ideal models for compound toxicity studies, but there is a need to accelerate basic and translational research in the field. Here, we developed an automated continuous imaging setup with the "read-on-ski" law of control to maximize temporal resolution with minimum culture plate vibration. High-accuracy performance was achieved: organoid screening and imaging were performed at a spatial resolution of 1.1 μm for the entire multi-well plate under 3 min. We used the in-house developed multi-well spinning device and cisplatin-induced nephrotoxicity model to evaluate the toxicity in kidney organoids using this system. The acquired images were processed via machine learning-based classification and segmentation algorithms, and the toxicity in kidney organoids was determined with 95% accuracy. The results obtained by the automated "read-on-ski" imaging device, combined with label-free and non-invasive algorithms for detection, were verified using conventional biological procedures. Taking advantage of the close-to-in vivo-kidney organoid model, this new development opens the door for further application of scaled-up screening using organoids in basic research and drug discovery.
Humans
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Kidney
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Organoids
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Pluripotent Stem Cells
3.Roles of detection, surveillance and early warning on outbreaks or epidemics of infectious diseases
Shukun WANG ; Shiwen ZHAO ; Xiaoqing FU ; Yinping MENG ; Yong ZHANG ; Chunrui LUO ; Yongming ZHOU ; Zhizhong SONG
Chinese Journal of Epidemiology 2021;42(5):941-947
Infectious diseases are still one of the leading causes of morbidity and death globally, affecting public health and life, social and economic development, and even national security. Early detection focuses on detecting the abnormal information of infectious disease outbreaks or epidemics in a timely and sensitive way to conduct field investigation and verification. It is also a precursor to effective surveillance and early warning system. The effective surveillance and early warning system can fully and accurately understand the real conditions, driving forces, and transmission chain of the occurrence of a specific infectious disease outbreak and epidemic and put forward scientific and effective prevention and control strategies and measures. Due to the measurement of the resources support and the particular data collection value, it is not easy to obtain epidemiological, etiological, and other data information in a timely, complete and accurate manner. This paper summarized the theory and technology on early detection, effective surveillance, and early warning information on infectious diseases. It also integrated and utilized the multi-source data, including effective infectious disease surveillance and the country's early warning system, to better understand the outbreak epidemic, causes, risks, processes, and driving forces. Thus, it is possible to set up a sensitive, specific staging measurement innovative technical system to monitor, early warning, and timely respond to acute infectious diseases through multidisciplinary cooperation in China. It provides the basis for strengthening the surveillance and early warning of new emerging and major infectious diseases and public health emergencies, avoiding the spread of inadequate response to infectious disease, and preventing the resources waste of over-response.
4.The detection of HLA-B27 gene and gene typing of ankylosing spondylitis patients by fluorescence PCR
Xiuxia LUO ; Zhihua YIN ; Chunrong ZHANG ; Jie CHEN ; Zhizhong YE ; Jinxian HUANG
International Journal of Laboratory Medicine 2018;39(13):1550-1552
Objective To investigate the clinical application value of fluorescence polymerase chain reaction (PCR) .in the human leucocyte antigen-B27(HLA-B27) gene and gene typing detection of ankylosing spondy-litis (AS) patients .Methods A total of 43 clinical blood samples of AS and 56 samples of healthy controls were collected in Shenzhen Futian hospital for rheumatic diseases from January 2014 to March 2015 .HLA-B27 gene was detected by flow cytometry .HLA-B27 gene and gene typing was also detected by the fluorescence PCR method .Results Among 43 samples ,40 samples were HLA-B27 positive(93 .02%) by flow cytometry while 39 samples were HLA-B27 positive (90 .70%) by fluorescence PCR .The total coincidence rate was 97 .50% .Among 39 positive samples ,32 samples were HLA-B2704 positive (82 .05%) and 7 samples were HLA-B2705 positive (17 .95%) .Conclusion The fluorescence PCR is an accurate method to detect HLA-B27 gene and presents high consistency with flow cytometry .It can also detect the HLA-B27 gene typing .It may have great clinical application value and prospects .
5.Anastomosis of dorsal branch of the proper digital nerve dorsal distal interphalangeal joint perforator flap to repair finger tip defect of skin tissue
Bangjun CHENG ; Zhaokuan ZHENG ; Zhizhong LIU ; Junwu HUANG ; Yi LUO
Clinical Medicine of China 2017;33(5):390-392
Objective To Summarize the application of digital artery dorsal distal interphalangeal joint perforator flap to repair finger tip skin defect,and at the same time,the method and effect of anastomosis of finger dorsal branch of digital nerve sensory reconstruction.Methods From September 2012 to March 2015,78 patients 92 fingers were treated in Orthopedics Department of Jinshan Branch Hospital of the Sixth People′s Hospital of Shanghai,all patients with finger artery distal interphalangeal joint dorsal cutaneous branches of the dorsal flap pedicled with retrograde,transferred to repair the skin defect of finger end,and anastomosis of dorsal branch of the proper digital nerve reconstruction.Results Postoperative vascular crisis occurred in 8 cases,2 cases of partial flap necrosis and healed after symptomatic treatment.All flaps survived,the wounds healed in I stage,and the donor site healed in I stage.All the 73 cases were followed up,the follow-up period ranged from 3.0 to 12.0 months,an average of (7.8±2.5) months.The postoperative appearance and feel good,soft texture,abrasion resistance,no tenderness,cold resistance,dynamic two-point recovered to 4.0-8.0 mm,average (5.3±0.9) mm.Static two-point discrimination was 4.0-9.0 mm,average (5.8±1.2) mm.The method of TAM was used to determine the function of the 67 fingers,good for the 7 finger,but also for the 5 finger and the difference of the 0 finger.The excellent and good rate was up to 93.7%.Conclusion The operation without sacrifice of major arteries and nerves by finger,the middle finger dorsal skin for non functional surface area,and at the same time by anastomosis of dorsal branch of the proper digital nerve reconstruction,without surgery two times,is one of the ideal surgical repair of skin defect of the finger end.
6.Posterior pilon fractures treated by buttress plating
Zhizhong WANG ; Mei HUANG ; Bin WANG ; Yepeng LIN ; Xinxu LI ; Xiaodong LUO ; Yanjun HU
Chinese Journal of Orthopaedic Trauma 2016;18(7):569-573
Objective To investigate the clinical efficacy of buttress plating for patients with posterior pilon fracture.Methods From April 2012 to January 2015,12 patients with posterior pilon fracture of the distal tibia were treated in our hospital.They were 7 men and 5 women,30 to 56 years of age (average,41.2 years).According to the CT classification by Haraguchi et al.,5 cases belonged to type I,3 to type Ⅱ and 4 to type Ⅲ.All the patients underwent open reduction and internal fixation with buttress plate via either a posterolateral approach or a dual posterolateral-posteromedial approach.All the patients were available for follow-up.The clinical outcomes were evaluated using the American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score and the visual analogue scale (VAS).The radiological evaluation was performed using the osteoarthritis-score (OA-score).Results The patients obtained an average follow-up of 21.2 months (range,from 12 to 30 months).Bone fractures united after an average of 15 weeks (range,from 13 to 19 weeks).The time for full weight walking averaged 16 weeks,ranging from 15 to 23 weeks.The ankle plantar flexion ranged from 36° to 42° (average,40.4°);the ankle dorsal extension ranged from 38° to 44° (average,42.6°).At the final follow-ups,the AOFAS scores ranged from 82 to 97 (average,88.2);the OA-score ranged from 0.6 to 0.8 (average,0.71);the VAS scores during rest,active motion and weight-bearing walking ranged from 0.5 to 0.8 (average,0.66),from 0.6 to 0.9 (average,0.82) and from 1.2 to 1.8 (average,1.41),respectively.No fracture malunion,implant loosening,pain or stiffness of the affected ankle was observed at the final follow-ups.Conclusion Buttress plating for posterior pilon fractures can lead to satisfactory clinical outcomes,because it ensures rigid fixation which in turn enables earlier postoperative mobilization.
7.miR-155 facilitates the differentiation of Th17 cells by inhibiting the gene expression of Ets-1
Zhihua YIN ; Xiuxia LUO ; Chunrong ZHANG ; Xinpeng CHEN ; Jinxian HUANG ; Zhizhong YE
Chinese Journal of Rheumatology 2015;19(11):730-734
Objective To elucidate the function way of micro RNA(miR)-155 in the differentiation of Th17 cells.Methods CD4+T cells were separated from mice spleens using MACS CD4+T cells separatinge kit and cultured with interleukins [interleukin (IL)-2, IL-23 and IL-6] which could induce CD4+ T cells differentiate into Th17 cells.IL-17 was detected by flow cytometry and enzyme linked immunosorbent assay (ELISA) after transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression levels of miR-155, IL-17A mRNA and Ets-1 mRNA were detected using fluorescent quantitation real-time quantitative polymerase chain reaction (RT-PCR).The si-Ets and miR-155 co-function for Th17 differentiation was analyzed.Data analysis was perfoemed using one-way analysis of variance (ANOVA) test and Dunnett test for pair-wise comparison and t test.P<0.05 was considered to be statistically significant.Results The CD4+T cells were divided into four groups (the untreated control untreat group, the treatment control treat group, the miR-155 mimnics group and miR-155 inhibitor group).IL-17 was scarcely expressed and secreted in the untreated control untreat group.The cells expression of IL-17 were significantly different among the four groups (F=160.549, P<0.01).The cells expressing of IL-17 were higher in the miR-155 mimics group (39.86±4.62)% than those at the miR-155 inhibitor group (22.02±2.81)%, P<0.01) and in the treated control treat group [(19.44±1.49)%, P<0.01].The level of IL-17 was also significantly different among the four groups (F=260.813, P<0.01).The level of IL-17 was higher in the miR-155 mimics group [(1 509±136) pg/ml] than that in the miR-155 inhibitor group [(923± 42) pg/ml, P<0.01);and in the treated control group [(767±94) pg/ml, P<0.01).The expression of miR-155 (12.53±0.80 vs 1.78±0.14, 7.16±0.62, 6.47±0.92, P<0.01) and IL-17A mRNA (46.55±6.71 vs 1.01±0.19,15.62±1.26, 14.20±2.73, P<0.01) was significantly higher than that in the other three groups, while the expression of Ets-1 mRNA was significantly lower (0.66±0.10 vs 1.19±0.04, 1.01±0.16, 1.37±0.27, P<0.01).si-Ets-2 was screened because it markedly inhibited the expression of Ets-1 mRNA among the three designed siRNAs.The expression of IL-17A mRNA was higher (17.19±3.58 vs 10.08±0.76, t=-3.361, P=0.028) and the expression of Ets-1 mRNA was lower (0.27±0.01 vs 0.74±0.03, t=-30.275, P<0.01) in si-Ets-2 group than that in si-Con group when si-Ets-2 or si-Con was co-transfected with miR-155 mimics or inhibitor lentiviral vectors.The expression of Ets-1 protein was lower in si-Ets-2 group than that in si-Con group by Western blotting and the decrease was markedly obvious in the miR-155 mimics group.Conclusion miR-155 can induce CD4+T cells to differentiate into Th17 cells by inhibiting the gene expression of Ets-1.
8.Expression and function of miRNA-16 in peripheral blood monouclear cells of systemic lupus erythematosus patients
Zhihua YIN ; Fenlian GUO ; Huiqiong ZENG ; Zhijun CAO ; Xiuxia LUO ; Zhizhong YE ; Hualin SUN
Clinical Medicine of China 2015;(2):97-99,100
Objective To investigate the expression of miRNA-16 in peripheral blood monouclear cells (PBMC)from systemic lupus erythematosus( SLE)patients. Methods Sixteen SLE patients who meet the diagnostic criteria of SLE revised in 1997 American rheumatology and 12 healthy individuals were selected as our subjects. Their peripheral blood were sampled. Total RNAs were extracted and purified. The level of miRNA-16 was determined by quantitative reverse transcription PCR( qRT-PCR). U6 was used as housekeeping control. The amount of target miRNA was normalized relative to the amount of U6(ΔCt =ΔCt miRNA-ΔCtU6 ). Relative expression levels were expressed as 2-ΔCt . Results The expression level of miRNA-16 in the SLE patients was 919. 87 ± 715. 45,significantly higher than that in the healthy control group(413. 6 3 ± 330. 69;t= -2. 497,P﹤0. 05). And miRNA-16 expression in SLE active group was 1 298. 79 ± 803. 79,significantly higher than that in SLE stable group(540. 95 ± 350. 15;t= -2. 445,P﹤0. 05). The level of miRNA-16 was related with AnuA (r=0. 669,P=0. 005),ESR(r=0. 608,P=0. 012)and SLEDAI(r=0. 530,P=0. 035). Conclusion The expression of miRNA-16 is high in SLE patients and it is related with SLE activity.
9.In vivo detection of severity of optic nerve crush using manganese-enhanced magnetic resonance imaging in rats.
Yun FENG ; ; Lisha LUO ; ; Zhizhong MA ; ; Xiaodong SUN ; Yuntao HU ;
Chinese Medical Journal 2014;127(3):522-527
BACKGROUNDTraumatic optic neuropathy (TON) is one of the reasons for permanent vision loss. Currently, the clinical practices may not be sufficient for direct assessments and comprehensively determining the location and extent of the patients with optic nerve injury in traumatic optic neuropathy. Magnetic resonance imaging (MRI) provides a non-invasive option. However, rare reports have found whether the differentdegree of injury of the optic nerve can be detected by manganese-enhanced MRI (MEMRI). This study aimed to explore the efficacy of MEMRI in the visual pathway for different severity of opitic nerve injury in rats.
METHODSThe different injuries of mild, moderate, and heavy damages were created by modified reverse tweezer and were evaluated by counting retinal ganglion cells (RGCs) and VEP ananlysis. Sprague-Dawley (SD) rats were intravitreally injected with 2 l of 25 mmol/L MnCl2, which has been confirmed as a safe injection concentration. The contrast-to-noise ratio (CNR) of MEMRI for optic nerve enhancement at different injury levels was measured.
RESULTSThe location of the significantly decreased signal point on optic nerve (ON) was corresponding to the location we made. However, similar findings are not obvious, or even have not been observed in 28 days in each group and also in 14 days at F100 group, indicating that MEMRI could be directly intuitive positioned in the early stage on the optic nerve injury.
CONCLUSIONSThe possibility of using MEMRI in optic nerve injury in a safe injection concentration of 25 mmol/L is confirmed. Therefore, it is possible to detect the severity of the optic nerve by MEMRI examination.
Animals ; Magnetic Resonance Imaging ; methods ; Male ; Manganese ; Optic Nerve Injuries ; diagnosis ; pathology ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; pathology
10.The expression of miR-155 and miR-146a in peripheral blood mononuclear cells and plasma of rheumatoid arthritis patients
Zhihua YIN ; Zhizhong YE ; Hualin SUN ; Jinxian HUANG ; Xiuxia LUO ; Fang CHEN ; Xingzhong HUANG ; Shuqiang MA
Chinese Journal of Rheumatology 2012;16(9):620-624
ObjectiveTo investigate the expression of miR-155 and miR-146a in peripheral blood mononuclear cells (PBMC) and plasma of rheumatoid arthritis (RA) patients.MethodsPBMC and plasma were separated from the peripheral blood of 34 RA patients and 15 healthy individuals.Total RNAs were isolated and miRNAs were purified.The levels of miR-155 and miR-146a were determined by quantitative reverse transcription PCR(qRT-PCR).U6 was used as housekeeping control.The amount of target miRNA was normalized relative to the amount of U6(ΔCt=ΔCtmiRNA-ΔCtU6).Relative expression levels were expressed as 2 △-ΔCt.Data were analyzed using SPSS 13.0 software.The test of homogeneity of variance and unpaired t-test was used to compare between groups.P values(2-tailed) less than 0.05 were considered as statistically significant.ResultsThe expressions of PBMC and plasma miR-155 were higher in RA patients than those in the healthy control individuals(0.08±0.08 vs 0.05±0.03,t=-2.225,P<0.05; 5.9±6.7 vs 1.3±2.0,t=-3.677,P<0.05).The expression of miR-146a in PBMC and plasma of RA patients and controls were(1.3±1.2 vs 0.8±0.6,t=-2.154,P<0.05)and(741±1001 vs 300±295,t=-1.669,P>0.05).According to their DAS28 value,RA patients were divided into high activity group (23 cases,DAS28≥5.0) and low disease activity group( 11cases,DAS28<5.0).The plasma miR-155 and miR-146a expressions were significantly higher in high activity group than those in low activity group.There were no significant differences in the expression of PBMC miR-155 and miR-146a between the two groups.ConclusionThe expression of PBMC and plasma miR-155 and miR-146a are higher in RA patients.The expression of plasma miR-155 and miR-146a are associated with RA patients' activity.Plasma miR-155 and miR-146a may be potential non-invasive biomarkers for RA diagnosis anddisease activity assessment.

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