1.Expression of porcine deltacoronavirus S protein in insect cells and preparation and identification of monoclonal antibody
Dantong LI ; Qing HE ; Dongliang WANG ; Beilei YU ; Qian YUAN ; Weijiao LIU ; Yi YANG ; Naidong WANG
Chinese Journal of Veterinary Science 2024;44(11):2309-2315
Porcine deltacoronavirus(PDCoV),a newly discovered virus in recent years,can cause severe diarrhea,vomiting,dehydration and even death in piglets.S protein is an important structur-al protein of PDCoV,which determines the host or tissue tropism of the virus,and is an important target for the development of PDCoV vaccines and detection methods.In order to prepare mono-clonal antibody(MAb)against the S protein of PDCoV,the recombinant plasmid of S protein was constructed based on the extracellular domain sequence of S protein of PDCoV epidemic strain in China and transformed into DH10Bac competent cells.The recombinant bacmid was identified by blue-white spot screening and PCR.BALB/c mice were immunized with S protein,and the spleen cells of immunized mice were fused with myeloma cells.The positive hybridoma cells that secreted stable antibodies were screened by indirect ELISA and subcloning.Five hybridoma cell superna-tants(MAb)specifically recognizing S protein(2E5,4D5,5D10,2F7 and 5A9)were identified by Western blot and immunofluorescence assay(IFA).Subsequently,the neutralization test showed that three of the monoclonal antibodies(2E5,4D5 and 5D10)could neutralize the virus to varying degrees.The S protein was successfully expressed and 5 monoclonal antibodies that can stably se-crete and specifically bind to PDCoV and S protein were prepared,which laid an important founda-tion for further research on the structure and function of S protein,the development of detection methods for PDCoV infection,and the prevention or treatment of PDCoV infection.
2.Application progress of multi-omics technology in clinical nursing research
Jianmei YANG ; Guiru LIN ; Wanyi OU ; Dongliang CHEN ; Yuanfei WANG ; Aiping WU ; Zumin CHEN ; Ze ZENG ; Xiaomin LU ; Chenli LIN ; Yinji LIANG
Chinese Journal of Nursing 2024;59(16):2044-2048,后插1
The update of multi-omics technology is a key means to promote the rapid development of accurate health model in the whole life cycle.It can formulate dynamic and accurate nursing measures and provide massive data information from the perspective of nursing biology of health and disease.At present,clinical nursing research faces many challenges such as insufficient application and transformation ability of multi-omics technology.This paper introduces the multi-omics technology,reviews the application status of multi-omics technology in cancer nursing,maternal and child nursing,chronic metabolic disease nursing and symptom management,and puts forward the cross integration and prospect of multi-omics technology and nursing research,so as to strengthen the information mining ability of nurses at different levels of health and disease,and provide an important basis for accelerating the clinical transformation of precision nursing.
3.Efficacy of enhanced recovery after surgery in laparoscopic treatment for children with Meckel's diverticulum
Xin FENG ; Xianwei ZHANG ; Fei ZHANG ; Yuan WEI ; Zhongyuan SUN ; Dongliang HOU ; Jushan SUN ; Quande FENG ; Yixi WANG ; Xingzhao CHEN
Chinese Journal of General Practitioners 2024;23(10):1079-1083
Clinical data of 166 children with Meckel's diverticulum, who were treated with laparoscopic surgery in our center from January 2015 to January 2023, were retrospectively analyzed, including 69 cases receiving enhanced recovery after surgery (ERAS group) and 97 cases with traditional perioperative care (control group). There were no significant differences in age ( t=1.391), gender ( χ2=1.067), body weight ( t=1.182 ), operation time ( t=1.093), diverticulum location ( Z=0.405), surgical procedures ( χ2=0.053), and intraoperative blood loss ( t=0.394) between two groups (all P>0.05). Compared to control group, ERAS group had shorter time for indwelling gastric tube (1.1±0.7 d vs.3.8±0.8 d), earlier postoperative feeding (2.5±0.6 d vs.4.9±0.7 d), less intravenous fluid infusion (3.9±1.0 d vs. 5.3±1.1 d), shorter length of hospital stay (8.2±1.6 d vs.10.9±2.3 d), and lower hospitalization expenditure (1.8±0.2)×10 4 yuan vs. (2.1±0.3)×10 4 yuan ( t=23.289,21.718,8.505,8.379,8.769,all P<0.05). There was no significant difference in incidence of postoperative complications between two groups ( χ2=0.431, P>0.05). The study indicates that patients treated with ERAS programmed laparoscopic Meckel's diverticulum surgery is safe and effective with rapid recovery and shorter hospital stay.
4.Study on the effect of low-frequency rTMS combined with NJF technique in improving the enhancement of muscular tone after stroke
Liru CUI ; Hanming WANG ; Aoran YANG ; Jian TAN ; Dongliang YANG ; Chunfang LI
China Medical Equipment 2024;21(10):90-95
Objective:To analyze the clinical effect of low-frequency repetitive transcranial magnetic stimulation(rTMS)combined with neuromuscular joint facilitation(NJF)technique in improving the enhancement of muscle tone after stroke.Methods:From October 2020 to October 22,a total of 114 patients with increased muscle tone at lower limb during the recovery period of stroke who admitted to Beijing Rehabilitation hospital were selected,and they were randomly divided into the observation group(57 cases)and the control group(57 cases)according to the random number table method.The control group received routine rehabilitation treatment.The observation group added low-frequency rTMS and NJF training on the basis of the routine rehabilitation treatment.The differences of clinical efficacy and various indicators of muscle tone of lower limb before and after treatment between two groups were compared,which included modified Ashwonh score(MAS)scale,functional ambulation category(FAC)scale of the motor of lower limb,Fugl-Meyer motor function scale-Lower limbs(FMA-L),clinical spasmodic index(CSI),modified Barthel index(MBI)of activity ability of daily life,integrated electromyogram(IEMG)and root mean square(RMS)of tibialis anterior muscle of lower limb in electromyogram.Results:After intervention,29 cases appeared significant effect,and 23 cases appeared valid result,and 5 cases appeared invalid result in the observation group,and 20 cases appeared significant effect,and 20 cases appeared valid result,and 17 cases appeared invalid result in the control group.The total effective rate of the observation group was 91.23%,which was significantly higher than 70.18%of the control group,and the difference was statistically significant(x2=8.111,P<0.05).Before intervention,there was no statistically significant difference in each indicator between the two groups(P>0.05).The MAS score,CSI score,IEMG and RMS of two groups after intervention were significantly lower than those before intervention,while the FAC score,FMA-L score and MBI score were significantly higher than those before intervention,and the differences were significant(t=3.377,4.398,5.554,P<0.05),respectively.At the same time,the MAS score,CSI score,IEMG and RMS of the observation group were significantly higher than those of the control group,and the differences were statistically significant(t=11.160,9.625,16.993,8.095,P<0.05),respectively.Conclusion:The combined application of low-frequency rTMS and NJF training can significantly improve the enhancement of muscle tone after stroke,and effectively reduce the muscle tone of lower limb,and improve the motor function of lower limb and the degree of spasticity,and improve the activity ability of daily life.
5.A trinity strategy for the treatment of multiple orthopedic trauma and assessment of its clinical application
Xiao CHEN ; Guangchao WANG ; Hao ZHANG ; Kaiyang LYV ; Qirong ZHOU ; Yunfei NIU ; Yan HU ; Yuanwei ZHANG ; Zuhao LI ; Hao SHEN ; Jin CUI ; Sicheng WANG ; Zhengrong GU ; Zhen GENG ; Dongliang WANG ; Zhehao FAN ; Shihao SHENG ; Chongru HE ; Jun FEI ; Yunfeng CHEN ; Haodong LIN ; Guohui LIU ; Zhiyong HOU ; Jiacan SU
Chinese Journal of Trauma 2024;40(10):888-896
Objective:To explore the clinical value of a trinity strategy for the treatment of multiple orthopedic trauma.Methods:A retrospective case series study was conducted to analyze the clinical data of 1 267 patients with multiple orthopedic trauma admitted to Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine and the First Affiliated Hospital of Navy Medical University from June 2013 to May 2023, including 862 males and 405 females, aged 18-93 years [(55.2±19.8)years]. Associated injuries included hemorrhagic shock in 632 patients, traumatic wet lung in 274, cranial injuries in 135, abdominal and pelvic bleeding in 116, pneumothorax in 89, urinary injury in 13, and vesical rupture in 8. All the patients were treated with the trinity strategy and the treatment process was divided into the phases of first aid, remodeling, and rehabilitation. The first aid phase focused on stabilizing symptoms and saving lives; the remodeling phase centered on restoring the anatomical structure and alignment; the rehabilitation phase aimed for functional recovery through the integration of both Western and traditional Chinese medicine. The all-cause mortality within 30 days after surgery and fracture healing time were calculated; the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, Hospital for Special Surgery (HSS) knee score and the American Orthopedic Foot & Ankle Society (AOFAS) ankle-hindfoot score at the last follow-up and the overall excellent and good rate of all joint function scores were measured. The short form health survey (SF-36) scores were collected preoperatively and at 6 months postoperatively, including 8 aspects such as physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health. The incidence of postoperative complications was recorded.Results:All the patients were followed up for 6-18 months [(10.2±4.2)months]. The mortality rate during the acute phase (within 30 days after surgery) was 2.37% with 12 deaths due to hemorrhagic shock, 10 due to traumatic brain injury, 6 due to multiple organ dysfunction syndrome (MODS), and 2 due to pulmonary infection. The average fracture healing time averaged 3.8-18 months [(11.5±4.2)months], with 89.49% of the patients having bone union within 12 months after surgery, 8.93% having bone union within 18 months after surgery, and 1.58% undergoing reoperation. For the patients with internal fixation failure and nonunion, the average healing time was extended to (10.2±2.2)months and (13.7±3.3)months respectively. At the last follow-up, the excellent and good rates of Constant-Murley shoulder score, Mayo elbow score, Gartland-Werley wrist score, Harris hip score, HSS knee score, and AOFAS ankle-hindfoot score were 83.93%, 90.24%, 94.12%, 85.57%, 88.46%, and 92.31% respectively, with an overall excellent and good rate of 89.11%. At 6 months after surgery, the SF-36 scores of all the patients in the eight dimensions,including the physical functioning, physical role, bodily pain, general health, vitality, social functioning, emotional role, and mental health were (74.4±8.6)points, (44.7±14.4)points, (77.4±10.9)points, (68.4±18.2)points, (72.5±16.0)points, (76.8±8.7)points, (49.9±17.6)points, and (72.8±17.9)points, significantly improved compared with those before operation [(63.4±12.7)points, (30.9±17.4)points, (56.4±18.0)points, (55.4±24.7)points, (53.5±21.0)points, (55.8±24.3)points, (36.9±24.0)points, (58.8±21.6)points] ( P<0.01). Complications of different degrees occurred in 214 patients (16.89%), including lung infections in 118 patients (9.31%), lower extremity deep vein thrombosis in 50(3.95%), pressure injuries in 26(2.05%), internal fixation failure in 12(0.95%), and nonunion in 8(0.63%). Conclusions:The trinity strategy provides whole-process management, personalized treatment, and overall rehabilitation for multiple orthopedic trauma. It can decrease mortality, shorten fracture healing time, improve joint function and quality of life, and reduce the incidence of complications.
6.Role of Ferroptosis in Osteoarthritis and Traditional Chinese Medicine Intervention: A Review
Xiaojing GUO ; Huan QIN ; Dongliang XIANG ; Yan WANG ; Li ZHANG ; Bo ZHANG ; Shujin WANG ; Xiaotong LI ; Mingyue ZHAO ; Shanhong WU ; Fei PEI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(19):263-272
Osteoarthritis (OA) is characterized by articular cartilage degeneration, synovial hyperplasia, hyperosteogeny, and narrowing of joint space, which can be caused by trauma, inflammation, and other factors. With the increasing global population aging, the incidence of OA is rising year by year, making it a major public health problem that urgently needs to be addressed. Exploring effective treatment schemes is particularly important. The pathogenesis of OA is complex, including oxidative stress, autophagy, and apoptosis. Recent studies have found that ferroptosis, a new type of cell death, is also an important pathogenic factor in OA, characterized by a series of complex changes such as iron ion accumulation, glutathione (GSH) depletion, and mitochondrial dysfunction. Research shows that inhibiting ferroptosis in chondrocytes can promote chondrocyte proliferation, delay extracellular matrix (ECM) degradation, and reduce synovial hyperplasia and inflammation. Targeting ferroptosis is a new direction in the treatment of OA. OA treatment includes intra-articular injections of steroids or hyaluronic acid and artificial joint replacement, but there are limitations. Traditional Chinese medicine (TCM) has been widely used in the treatment of various diseases because of its low cost, low drug resistance, and few side effects. Cell and animal experiments have further confirmed that TCM can intervene in the treatment of OA with ferroptosis from multiple targets, multiple levels, and aspects, but the mechanism of its treatment of OA based on ferroptosis has not been clarified. This paper discussed iron metabolism, lipid peroxidation, cysteine/glutamate transporter system Xc- (system Xc-)/GSH/glutathione peroxidase 4 (GPX4) pathway, nicotinamide adenine dinucleotide phosphate(NADPH)/ferroptosis suppressor protein 1 (FSP1)/coenzyme Q10 (CoQ10) pathway, tumor protein p53 in OA, and related molecular targets of Chinese medicine monomers and compounds on ferroptosis inhibition. Their potential therapeutic mechanisms were further analyzed to provide theoretical guidance for the treatment of OA by TCM and useful reference for the research and development of related drugs.
7.Expert consensus on cryoablation therapy of oral mucosal melanoma
Guoxin REN ; Moyi SUN ; Zhangui TANG ; Longjiang LI ; Jian MENG ; Zhijun SUN ; Shaoyan LIU ; Yue HE ; Wei SHANG ; Gang LI ; Jie ZHNAG ; Heming WU ; Yi LI ; Shaohui HUANG ; Shizhou ZHANG ; Zhongcheng GONG ; Jun WANG ; Anxun WANG ; Zhiyong LI ; Zhiquan HUNAG ; Tong SU ; Jichen LI ; Kai YANG ; Weizhong LI ; Weihong XIE ; Qing XI ; Ke ZHAO ; Yunze XUAN ; Li HUANG ; Chuanzheng SUN ; Bing HAN ; Yanping CHEN ; Wenge CHEN ; Yunteng WU ; Dongliang WEI ; Wei GUO
Journal of Practical Stomatology 2024;40(2):149-155
Cryoablation therapy with explicit anti-tumor mechanisms and histopathological manifestations has a long history.A large number of clinical practice has shown that cryoablation therapy is safe and effective,making it an ideal tumor treatment method in theory.Previously,its efficacy and clinical application were constrained by the limitations of refrigerants and refrigeration equipment.With the development of the new generation of cryoablation equipment represented by argon helium knives,significant progress has been made in refrigeration efficien-cy,ablation range,and precise temperature measurement,greatly promoting the progression of tumor cryoablation technology.This consensus systematically summarizes the mechanism of cryoablation technology,indications for oral mucosal melanoma(OMM)cryotherapy,clinical treatment process,adverse reactions and management,cryotherapy combination therapy,etc.,aiming to provide reference for carrying out the standardized cryoablation therapy of OMM.
8.Preliminary identification of the cloning, expression, and function of Marmota himalayana type I interferon receptor β subunit
Ying TAO ; Dongliang YANG ; Baoju WANG ; Yi LIU ; Wenjia GUI ; Zhi LI ; Hebin FAN
Journal of Clinical Hepatology 2024;40(2):278-283
ObjectiveTo clone the gene of Marmota himalayana type Ⅰ interferon receptor β subunit (mhIFNAR2), and to perform antibody preparation and functional identification. MethodsRT-PCR was used for amplification in the spleen tissue of Marmota himalayana to obtain the sequence, which was cloned to the prokaryotic expression vector pRSET-B to express the recombinant protein. Electrophoresis and Western blot were used for identification. BALB/c mice were immunized with the recombinant protein to prepare the polyclonal antibody of its extracellular domain; immunohistochemistry, immunofluorescence assay, and Western Blot were used for identification, and the method of siRNA blockade was used to investigate its function. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for comparison between two groups. ResultsA fragment of mhIFNAR2 (149 — 1 300 bp) was obtained from spleen tissue, which showed the highest homology of 98.05% in marmot. A prokaryotic expression plasmid was successfully constructed for expression of the extracellular domain of the mhIFNAR2(50-181aa) and was named pRSET-B.mhIFNAR2, and the recombinant protein expressed by this plasmid had a molecular weight of 27 kD, a purity of about 95% after purification, and a concentration of 160 μg/mL. After BALB/c mice were immunized with the purified recombinant protein, 1∶1 000 specific polyclonal antibodies were obtained, and immunohistochemistry and immunofluorescence assay showed the expression in cell membrane and cytoplasm. Among the three siRNAs synthesized, the siRNA starting from the 277 locus (siRNA277) could silence the expression of target genes and weaken the interferon signaling pathway compared with the blank control group and the negative control group (both P<0.05). ConclusionThe fragment of mhIFNAR2 is obtained, and the polyclonal antibody for the extracellular domain of mhIFNAR2 is successfully prepared, with relatively high titer and specificity, and can be used for immunohistochemistry, immunofluorescence assay, and Western blot.
9.Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine (version 2024)
Xiao CHEN ; Hao ZHANG ; Man WANG ; Guangchao WANG ; Jin CUI ; Wencai ZHANG ; Fengjin ZHOU ; Qiang YANG ; Guohui LIU ; Zhongmin SHI ; Lili YANG ; Zhiwei WANG ; Guixin SUN ; Biao CHENG ; Ming CAI ; Haodong LIN ; Hongxing SHEN ; Hao SHEN ; Yunfei ZHANG ; Fuxin WEI ; Feng NIU ; Chao FANG ; Huiwen CHEN ; Shaojun SONG ; Yong WANG ; Jun LIN ; Yuhai MA ; Wei CHEN ; Nan CHEN ; Zhiyong HOU ; Xin WANG ; Aiyuan WANG ; Zhen GENG ; Kainan LI ; Dongliang WANG ; Fanfu FANG ; Jiacan SU
Chinese Journal of Trauma 2024;40(3):193-205
Osteoporotic proximal humeral fracture (OPHF) is one of the common osteoporotic fractures in the aged, with an incidence only lower than vertebral compression fracture, hip fracture, and distal radius fracture. OPHF, secondary to osteoporosis and characterized by poor bone quality, comminuted fracture pattern, slow healing, and severely impaired shoulder joint function, poses a big challenge to the current clinical diagnosis and treatment. In the field of diagnosis, treatment, and rehabilitation of OPHF, traditional Chinese and Western medicine have accumulated rich experience and evidence from evidence-based medicine and achieved favorable outcomes. However, there is still a lack of guidance from a relevant consensus as to how to integrate the advantages of the two medical systems and achieve the integrated diagnosis and treatment. To promote the diagnosis and treatment of OPHF with integrated traditional Chinese and Western medicine, relevant experts from Orthopedic Expert Committee of Geriatric Branch of Chinese Association of Gerontology and Geriatrics, Youth Osteoporosis Group of Orthopedic Branch of Chinese Medical Association, Osteoporosis Group of Orthopedic Surgeon Branch of Chinese Medical Doctor Association, and Osteoporosis Committee of Shanghai Association of Integrated Traditional Chinese and Western Medicine have been organized to formulate Expert consensus on the diagnosis and treatment of osteoporotic proximal humeral fracture with integrated traditional Chinese and Western medicine ( version 2024) by searching related literatures and based on the evidences from evidence-based medicine. This consensus consists of 13 recommendations about the diagnosis, treatment and rehabilitation of OPHF with integrated traditional Chinese medicine and Western medicine, aimed at standardizing, systematizing, and personalizing the diagnosis and treatment of OPHF with integrated traditional Chinse and Western medicine to improve the patients ′ function.
10.Advances in effect mechanism research of traditional Chinese medicine on comorbid depressive symptoms in chronic pain
Guanghua LIU ; Dongliang SHI ; Wen FU ; Zihua WANG ; Jiakang CUI
China Pharmacy 2024;35(19):2438-2443
Chronic pain refers to pain that persists or recurs for more than 3 months and has a high comorbidity rate with emotions such as depression and anxiety. The complex pathogenesis of comorbid depressive symptoms in chronic pain poses great challenges to clinical diagnosis and treatment. This article reviews the research progress on the effect mechanism on the comorbid depressive symptoms in chronic pain by traditional Chinese medicine monomers, compound formulas, and acupuncture therapy in recent years. It is found that traditional Chinese medicine monomers (flavonoids, phenols, terpenes, coumarins, alkaloids), traditional Chinese medicine formulas (Aconitum carmichaeli decoction, Chaihu guizhi decoction, etc.), and acupuncture therapy (acupuncture at acupoints such as Baihui, Yintang, Hegu, and Taichong) can effectively improve comorbid depressive symptoms in chronic pain by regulating the levels of neurotransmitters, brain-derived neurotrophic factors and inflammatory factors, and the activity of glial cells in the central nervous system.

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