Single-cell RNA sequencing technology takes a single cell as research object,counts and analyzes the gene expres-sion level of each transcript and heterogeneity between cells.This technology makes up for the defects of traditional sequencing techno-logy to some extent.Autoimmune disease is the damage or dysfunction of autotissue cells caused by autoimmune tolerance or abnormal regulation of autoimmunity cells.In this paper,the research results on application of single-cell RNA sequencing technology in autoim-mune disease in recent years are reviewed,which provides valuable clues for early realization of precise medical treatment.

A 11-year old female patient with severe thalassemia, receipt a lentivirus-based cell and gene therapy (CGT) therapy in Shenzhen Children′s Hosptial on July 27th, 2021. At the two follow-up visits after discharge, patient were continuously tested positive for HIV screening through HIV Ag/Ab Combo assay (chemiluminescence Immunoassay), and the viral load results of HIV-1 nucleic acid testing (NAT) were both>5 000 copies/ml. The patient can be diagnosed with HIV infection according to the National Guideline for Detection of HIV/AIDS(2020 Revised Edition). The thorough investigation findings and supplementary experiment results indicated that the false-positive HIV-1 NAT results was caused by cross-reactivity between the target sites detected by conventional HIV-1 NAT reagents and the lentiviral vectors fragments integrated into the genome of patient′s hematopoietic stem/progenitor cells. In conclusion, it is important for laboratories to select appropriate HIV-1 NAT testing platforms which won′t cause cross-reactivity for the testing of samples from patients who have been treated with HIV-derived vectors. It is also recommended to design and develop NAT testing platforms with multiple target regions labeled by different fluorescents for HIV NAT supplementation experiment to reduce the risk of false-positive diagnoses of HIV infection.

Objective:To describe the clinical features of a patient of anti-neurofascin 186 (NF186) antibody associated acute immune sensory polyradiculopathy (AISP), and enhance understanding of AISP/chronic immune sensory polyradiculopathy (CISP).Methods:The clinical characteristics, diagnosis and treatment of a domestic AISP patient with NF186 antibody positive admitted to the First Hospital of Shanxi Medical University in December 2021 were summarized, and the previously reported cases of AISP/CISP were systematically reviewed.Results:The patient was a 62-year-old male with acute onset. The clinical manifestations included severe sensory ataxia, increased protein in cerebrospinal fluid, no response to stimulation of the central segment of somatosensory evoked potentials (SEP), normal sensory and motor nerve conduction, and positive serum anti-NF186 antibody (1∶32). After glucocorticoid treatment, the clinical symptoms and SEP were significantly improved. The drug was stopped for 2 months, and there was no recurrence. There were 23 cases of AISP and CISP with complete data reported in the literature (including this patient). The age of onset was (54.7±17.7) years, and the ratio of male to female was 1.88. Three patients with acute onset were classified as AISP. A total of 95.7% (22/23) of patients showed sensory ataxia without limb weakness, 95.0% (19/20) of patients showed prolonged cortical potential latency or even no response, and 95.5% (21/22) of patients showed increased cerebrospinal fluid protein in varying degrees, and nerve root thickening or abnormal enhancement was not common. All 10 patients receiving immunotherapy responded to corticosteroids or intravenous immune globulin. Only 6 AISP/CISP articles reported screening for anti-ganglioside antibodies or Ranvier′s node-paranodal region-related antibodies, and no positive NF186 antibodies were reported. All the 3 patients with AISP had some characteristics of CISP/chronic inflammatory demyelinating polyradiculoneuropathy, and there was no significant difference between AISP and CISP patients in clinical features except the mode of onset.Conclusions:NF186 antibody could cause AISP, which presents as acute onset sensory ataxia. AISP is responsive to glucocorticoid therapy. Except for the mode of onset, AISP and CISP are difficult to distinguish from clinical, electrophysiological, pathological aspects and pathogenic antibodies, so they may be two different manifestations of the same disease.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine

Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

Endoscopic data of 108 upper gastrointestinal elevated lesions caused by vascular or hemangioma compression by endoscopic ultrasonography (EUS) at the Second Affiliated Hospital of Soochow University, Changshu No.1 People's Hospital, Kushan Hospital of Chinese Medicine and Traditional Chinese Medicine Hospital of Changshu from December 2010 to June 2019 were retrospectively summarized. The results showed that lesions were mainly located in the esophagus [50.9% (55/108)] and stomach [47.2% (51/108)], especially in the middle [40.0% (22/55)] and upper esophagus [36.4% (20/55)], body [66.7% (34/51)] and fundus of stomach [31.4% (16/51)], respectively. The major etiology included splenic artery and aneurysm compression [29.6% (32/108)], aortic compression [23.1% (25/108)], isolated esophageal venous aneurysm compression [13.9% (15/108)] and gastric submucosal vein and venous aneurysm compression [12.0% (13/108)], with diverse endoscopic presentation. The above results suggest that elevated lesions of upper gastrointestinal tract caused by blood vessels and hemangiomas are mostly due to external vascular pressure outside the lumen, but ectopic submucosal arteries and isolated phlebangioma are not uncommon. The lesions are widely distributed with different gastroscopic manifestations. EUS is important for definite diagnosis, and can be combined with color Doppler technique, CT plain scan and angiographic reconstruction if necessary.

This study probed the protective effect of recombinant

OBJECTIVE: To study the protective effect of enhanced peroxisome proliferator activated receptor γ (PPARγ) pathway against apoptosis of long-term cultured primary nerve cells. METHODS: A natural aging model was established in primary rat nerve cells by long-term culture for 22 days. The cells were divided into control group, 0.1, 1.0, 5.0, and 10 μmol/L GW9662 intervention groups, and 0.1, 1.0, 5.0, and 10 μmol/L pioglitazone intervention groups. The cell viability was assessed using MTT assay and the cell morphological changes were observed after the treatments to determine the optimal concentrations of GW9662 and pioglitazone. Double immunofluorescence labeling and flow cytometry were used to observe the changes in the number of viable cells and cell apoptosis following the treatments; immunocytochemical staining was used to assess the changes in the anti-oxidation ability of the treated cells. RESULTS: The optimal concentrations of GW9662 and pioglitazone determined based on the cell viability and morphological changes were both 1 μmol/L. Compared with the control group, GW9662 treatment significantly lowered while pioglitazone significantly increased the total cell number and nerve cell counts ( < 0.05), and nerve cells in the cell cultures maintained a constant ratio at about 80% in all the groups ( > 0.05). GW9662 significantly enhanced while pioglitazone significantly lowered the cell apoptosis rates compared with the control group ( < 0.05). GW9662 obviously lowered SOD activity and GSH content in G group ( < 0.05) and increased MDA content in the cells ( < 0.05), and pioglitazone resulted in reverse changes in SOD, GSH and MDA contents in the cells ( < 0.05). CONCLUSIONS Activation of PPARγ pathway protects long-term cultured primary nerve cells by enhancing cellular anti-oxidant capacity and reducing cell apoptosis, suggesting a potential strategy for anti-aging treatment of the nervous system through intervention of the PPARγ pathway.
Anilides ; administration & dosage ; pharmacology ; Animals ; Apoptosis ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cellular Senescence ; physiology ; Neurons ; cytology ; PPAR gamma ; metabolism ; Pioglitazone ; administration & dosage ; pharmacology ; Rats

Objective To investigate the cognition and attitude toward integrating old-age care with medical services among the elderly,nurses from medical institutions and old-age care institution staffs,in order to provide references for developing implementation strategies of integrating old-age care with medical service.Methods The convenience sampling method was conducted,and 963 people.from 4 cities in Henan Province were enrolled in this survey from July to September 2016,including 599 elderly people,248 nurses from medical institutions and 166 old-age care institution staffs.All volunteers were investigated by using self-designed questionnaires.Results A total of 1 000 questionnaires were distributed,and 963 valid questionnaires were retrieved.The effective recovery rate was 96.3％.The cognition of integrating old-age care with medical service in all volunteers was low.There were differences in the needs of all nursing care services in the integrative old-age care and medical services,the needs of Chinese traditional medical care services were low.The highest demand of service capabilities was skills of coping urgent issues,while the lowest demand of service capabilities was education levels of care givers;the scores on evaluation of the two demands in the elderly were (4.803± 0.453) and (3.124± 1.246) points,those in the old-age care institution staffs were (4.819±0.468) and (2.698± 1.346) points,there was no statistically significant difference between the two groups (P＞0.05).Conclusion In the progress of promoting the integration of old-age care and medical services,it is necessary pay more attention on the promotion among non-urban elderly people,and improving living facilities as well as the quality of care services.Integrated management and hierarchical training for nursing professionals and old-age care institutions staffs should be carried out gradually.

Objective:To explore the correlation between cerebrovascular hemodynamic index (CVHI) accumulative score and subclinical arteriosclerosis indicators.Methods:A total of 27 184 cases were collected from the Health Management Center,the Third Xiangya Hospital,Central South University.Linear regression analysis was carried out to confirm the correlations between CVHI accumulative score and the modified Framingham stroke profile (FSP),as well as between CVHI accumulative score and cerebrovascular diseases (ICVD) scale.The correlation between CVHI accumulative score and brachial-ankle pulse wave velocity (baPWV),carotid plaque orcarotid intima-media thickness (CIMT) was analyzed by multifactor logistic regression model in 11 580 cases.Moreover,the correlation between CVHI accumulative score and microalbuminuria or serum cystatin C was performed by multifactor logistic regression model in 9 860 cases.Results:In this study,the people whose CVHI accumulative score was less than 75 accounted for 12.98％.The CVHI accumulative score was negatively related with the modified FSP score (r=-0.484,P＜0.01) or ICVD score (r=-0.455,P＜0.01).The multifactor logistic regression model found that the baPWV,carotid plaque,microalbuminuria and serum cystatin C were independent predictors for CVHI accumulative score.Conclusion:The CVHI accumulative score is correlated with the modified FSP score,ICVD score and indexes of subclinical arteriosclerosis (baPWV,carotid plaque,microalbuminuria and serum cystatin C).The CVHI accumulative score could be used as a tool for zero-level and primary prevention of cerebral stroke.