1.The mechanism of SAP overexpression in alleviating periodontitis in mice
HUANG Yinyin ; LIANG Dongliang ; ZOU Yaokun ; HAN Jingru ; GE Qing ; LIU Xueyan ; GUO Yadong ; HUANG Xinli ; YANG Lan
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(8):619-630
Objective:
To investigate the mechanism by which serum amyloid P component (SAP) alleviates periodontitis in mice, providing an experimental basis to establish SAP as a novel therapeutic agent for periodontitis.
Methods:
Ethical approval was obtained from the Institutional Animal Ethics Committee. Periodontitis models were established in wild-type (WT) mice and SAP-transgenic (SAP-Tg) mice, divided into four groups: WT control (WT group), WT periodontitis (WT+P group), SAP-Tg control (Tg group), and SAP-Tg periodontitis (Tg+P group). On day 7, the mice were euthanized, and periodontal tissues, teeth, and alveolar bone were collected. SAP protein expression was detected by enzyme-linked immunosorbent assay (ELISA). Micro-CT and HE staining were used to measure alveolar bone resorption (distance from the cementoenamel junction to the alveolar bone crest). Tartrate-resistant acid phosphatase (TRAP) staining was performed to assess osteoclast number, and immunohistochemistry (IHC) was employed to evaluate macrophage infiltration. The expression levels of inflammatory cytokines including interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were measured by qRT-PCR. Oral microorganism composition was analyzed using 16S ribosomal RNA (16S rRNA) gene sequencing. Additionally, macrophages from WT and SAP-Tg mice were isolated to establish an in vitro inflammation model, divided into WT+LPS and Tg+LPS groups. The expression of macrophage polarization-related genes including inducible nitric oxide synthase (iNOS), CD86, CD163, and CD206) were assessed by qRT-PCR. After the induction of osteoclast differentiation, TRAP staining was performed.
Results:
ELISA results demonstrated that periodontal tissues from Tg+P group mice exhibited higher levels of SAP expression compared to the WT+P group. Micro-CT and HE staining analyses revealed that the Tg+P group showed reduced alveolar bone resorption, indicated by a shorter distance between the cementoenamel junction and alveolar bone crest, compared to the WT+P group. Furthermore, TRAP staining results indicated a decrease in osteoclast numbers in the Tg+P group compared to the WT+P group. IHC and qRT-PCR results indicated reduced macrophage infiltration and decreased expression of IL-1β, IL-6, and TNF-α in the Tg+P group. Oral microorganism sequencing showed no significant difference in periodontitis-associated pathogenic bacteria between WT+P and Tg+P groups. In vitro experiments demonstrated that compared to the WT+LPS group, the Tg+LPS group exhibited downregulated M1 macrophage markers (iNOS and CD86) and upregulated M2 macrophage markers (CD163 and CD206). TRAP staining confirmed fewer osteoclasts in the Tg+LPS group.
Conclusion
SAP overexpression effectively alleviates periodontitis severity in mice by inhibiting M1 macrophage polarization, reducing pro-inflammatory cytokine expression, and suppressing osteoclast differentiation, thereby attenuating alveolar bone resorption.
2.Research progress of MAVS in viruses evading host natural immunity
Donglin BI ; Xiaoli YANG ; Dongliang YANG ; Fangcheng LIU ; Xiaowen ZHANG ; Qiongyi LI ; Jialin BAI
Chinese Journal of Immunology 2024;40(11):2452-2457,2464
Innate immunity is the first line of the host cell defensing against viral infection,in which the pattern recognition re-ceptors(PRRs)such as Toll-like receptors(TLRs)and retinoic acid induces gene Ⅰ-like receptors(RLRs)play an important role.After virus infection,mitochondrial antiviral signaling protein(MAVS)in PRRs-mediated signaling pathway,which are of the com-mon linker molecule for downstream signal transmission,can receive signals transmitted by upstream TLR and RIG-Ⅰ,activate down-stream NF-κB and IRF3/7 signaling pathways leading to the activation of interferon(IFN)expression.Therefore MAVS acts as a bridge in the innate immune signaling pathway.More and more studies have shown that viruses have evolved a series of mechanism to escape the innate immune response over the long course of their evolution,and evaded the host's antiviral immune response by interfer-ing with multiple sites in the MAVS-mediated signaling pathway so as to complete its own replication and proliferation.In this paper,the role of MAVS in IFN-Ⅰ pathway and its latest research progress in the mechanism of anti-DNA viruses and anti-RNA viruses reac-tion are reviewed,providing theoretical basis for further studying the detailed mechanism of anti-virus of MAVS.
3.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.
4.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.
5.Effect and mechanism of sodium hydrosulfide on cerebral cortical injury in rats with cerebral ischemia-reperfusion
Yinghong LI ; Yue ZUO ; Kunli YANG ; Yankai REN ; Dongliang LI
Journal of Xinxiang Medical College 2024;41(10):904-910
Objective To explore the effect and mechanism of sodium hydrosulfide(NaHS)on cerebral cortical injury in rats with cerebral ischemia-reperfusion.Methods A total of 80 male SD rats were divided into a sham operation group,an NaHS group,an ischemia-reperfusion group,and an ischemia-reperfusion+NaHS group using a random number table method,with 20 rats in each group.The rats in the ischemia-reperfusion group and ischemia-reperfusion+NaHS group were used to prepare ischemia-reperfusion models.The rats in the ischemia-reperfusion+NaHS group were given 50 μmol·kg-1 NaHS intraperitoneally after cerebral ischemia,while the rats in the ischemia-reperfusion group were given an equal volume of physiological saline intraperitoneally after cerebral ischemia.The rats in the sham operation group and NaHS group only had vessels isolated but not occluded with threads.The rats in the NaHS group were given 50 μmol·kg-1 NaHS intraperitoneally after vascular detachment,while the rats in the sham operation group were given an equal volume of physiological saline intraperitoneally after vascular detachment.Twenty-four hours after modeling,the neurological deficits of rats in the four groups were evaluated by using the modified neurological severity score(mNSS),the cerebral infarction of rats in the four groups was observed aftertriphenyltetrazolium chloride staining,the levels of pro-inflammatory cytokines interleukin-1β(IL-1β)and interleukin-18(IL-18)in theischemiccerebralcortical tissues of rats in the four groups were detected by using enzyme-linked immunosorbent assay,the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)in the ischemic cerebral cortical tissues of rats in the four groups was detected by using Western blot,and the activity of microglia in the ischemic cerebral cortical tissues of rats in the four groups was detected by using immunofluorescence.Results The brain tissues of rats in the sham operation group and the NaHS group showed uniform orange red color,while the rats in the ischemia-reperfusion group and the ischemia-reperfusion+NaHS group had pale infarcted areas in the ischemic cerebral cortex and striatum.The cerebral infarction area of rats in the ischemia-reperfusion group and the ischemia-reperfusion+NaHS group were significantly larger than that in the sham operation group and the NaHS group,while the cerebral infarction area of rats in the ischemia-reperfusion+NaHS group was significantly smaller than that in the ischemia-reperfusion group(P<0.05).The mNSS scores of rats in the ischemia-reperfusion group and the ischemia-reperfusion+NaHS group were significantly higher than those in the sham operation group and the NaHS group,while the mNSS score of rats in the ischemia-reperfusion+NaHS group was significantly lower than that in the ischemia-reperfusion group(P<0.05).The relative expression levels of NLRP3 protein in the ischemic cerebral cortex of rats in the ischemia-reperfusion group and the ischemia-reperfusion+NaHS group were significantly higher than those in the sham operation group and the NaHS group,while the relative expression level of NLRP3 protein in the ischemic cerebral cortex of rats in the ischemia-reperfusion+NaHS group was significantly lower than that in the ischemia-reperfusion group(P<0.05).The levels of IL-1β and IL-18 in the ischemic cerebral cortex of rats in the ischemia-reperfusion group and the ischemia-reperfusion+NaHS group were significantly higher than those in the sham operation group and the NaHS group,while the levels of IL-1β and IL-18 in the ischemic cerebral cortex of rats in the ischemia-reperfusion+NaHS group were significantly lower than those in the ischemia-reperfusion group(P<0.05).The rats in both sham operation group and NaHS group showed normal morphology of microglia in the ischemic cerebral cortical area.Compared with the sham operation group and the NaHS group,the rats in the ischemia-reperfusion group showed a significant reduction in the protrusion of microglia in the ischemic cerebral cortical area and a significant increase in the brightness of microglial cell bodies,and the cells appeared as clumps,losing their normal morphology,with a higher proportion of activated microglia.Compared with the ischemia-reperfusion group,the rats in the ischemia-reperfusion+NaHS group showed an increase in the number of microglial processes in the ischemic cerebral cortex,with fewer activated microglia and significantly improved morphology.The total numbers of microglia in the ischemic cerebral cortex of rats in the ischemia-reperfusion group and theischemia-reperfusion+NaHS group were significantly lower than those in the sham operation group and the NaHS group,while the total number of microglia in the ischemic cerebral cortex of rats in the ischemia-reperfusion+NaHS group was significantly higher than that in the ischemia-reperfusion group(P<0.05).Conclusion NaHS can significantly improve brain injury in ischemia-reperfusion rats by inhibiting the inflammatory response and microglial activation induced by NLRP3 and reducing the levels of pro-inflammatory cytokines IL-1β and IL-18.
6.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.
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.Clinical observation of free composite tissue flap transplantation in repairing head skin defect with artificial dural exposure infection
Zhigang XU ; Dongliang ZHANG ; Ting ZHANG ; Ting HE ; Jingtao WEI ; Qiaohua CHEN ; Xuekang YANG
Journal of Chinese Physician 2024;26(3):349-353
Objective:To explore the effect of free transplantation of composite tissue flap from the anterior lateral aspect of the femur in repairing head skin defects with artificial dural exposure infection.Methods:A retrospective study was conducted on 13 patients admitted to the First Affiliated Hospital of Air Force Military Medical University from April 2018 to August 2020 with craniotomy complications, including craniotomy skin and soft tissue defects combined with artificial dural exposure and infection. After preoperative anti infection treatment, the neurosurgery department participated in debridement and removed the artificial dura mater as much as possible during the operation. A composite tissue flap carrying the fascia lata was designed for the anterior lateral aspect of the thigh, and the flap artery and vein were anastomosed with the superficial temporal artery and superficial temporal vein/middle temporal vein respectively. The defect of the dura mater was repaired with the fascia lata with blood supply. The flap was used to seal the wound, and the donor site was directly sutured or transplanted with autologous medium thick skin graft. The postoperative blood supply and survival of the flap, the presence of cerebrospinal fluid leakage, and the healing of the donor site were observed; The observation of dural integrity and postoperative effects of skull reconstruction using cranial magnetic resonance imaging was followed up.Results:Among the 13 patients in this group, 11 patients had their artificial dura mater completely removed, while 2 patients were not completely removed due to severe adhesion. Among them, 1 patient had a residual area of 0.8 cm×1 cm, and the other had 3 residual areas, with a maximum area of 0.5 cm×0.7 cm; All transplanted skin flaps survived, with 12 cases achieving primary healing and 1 case of partial wound rupture after suture removal, which healed after conservative dressing change; All patients had no cerebrospinal fluid leakage; There was one case of partial necrosis of the graft in the donor site, which healed after supplementing the graft; Thirteen patients underwent cranial magnetic resonance imaging at 3-6 months postoperatively, all of which showed intact dura mater; Among them, 8 patients have completed skull reconstruction surgery, and all of them have healed well after reconstruction, with a good appearance of the surgical area.Conclusions:For wounds with head skin defects and exposed artificial dura mater infection, free transplantation of the anterior lateral composite tissue flap carrying the fascia lata can effectively cover the wound and repair the dura mater defect, achieve good function and appearance, and create favorable conditions for later skull reconstruction.
10.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.


Result Analysis
Print
Save
E-mail