BACKGROUND:Anthocyanin is one of the most important active components in Lycium ruthenicum Murr,which has antioxidant and immunomodulatory effects.CD146+human adipose-derived pericytes/perivascular cells(CD146+hAD-PCs)are the progenitors of bone marrow mesenchymal stem cells,which can promote the proliferation and differentiation of hematopoietic stem/progenitor cells in vitro.The support effect of anthocyanin in combination with CD146+hAD-PCs on umbilical cord blood hematopoietic stem/progenitor cells remains to be studied. OBJECTIVE:To investigate the supporting effect of anthocyanins in Lycium ruthenicum Murr(ALRM)combined with CD146+hAD-PCs on umbilical cord blood CD34+hematopoietic stem/progenitor cells(UCB CD34+HSPCs)in vitro. METHODS:The CCK-8 assay was used to detect the effect of different concentrations(0,200,400,600,800,1 000 mg/L)of ALRM on the proliferation of CD146+hAD-PCs.Flow cytometry was used to detect the effect of ALRM on the cell cycle of CD146+hAD-PCs.The co-culture experiments were divided into blank group,ALRM group,CD146+hAD-PCs group,and ALRM+CD146+hAD-PCs group to analyze the in vitro supporting effect of ALRM combined with CD146+hAD-PCs on UCB CD34+HSPCs.The number of expanded cells and the number of colony-forming units were compared at 1,2,and 4 weeks of co-culture.The immunophenotype of cells was detected by flow cytometry.The level of cytokines was detected by enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:(1)The cell viability of CD146+hAD-PCs was highest at an ALRM concentration of 200 mg/L,the proportion of G0/G1 phase cells decreased and the proportion of S and G2/M phase cells increased in CD146+hAD-PCs(P<0.01).(2)The change in number of UCB CD34+HSPCs cells in the ALRM+CD146+hAD-PCs group was higher than that in the ALRM group at 1,2,and 4 weeks of co-culture(all P<0.05),and higher than that in CD146+hAD-PCs group at 2 and 4 weeks of co-culture(all P<0.05).The number of cells in the ALRM group and blank group decreased gradually with the extension of co-culture time.(3)Colony forming capacity and immunophenotype analysis:The number of colony-forming units in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group and ALRM group at 1 and 2 weeks of co-culture(P<0.05).The proportion of CD45+and CD34+CD33-cells in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group at 1 and 2 weeks of co-culture(all P<0.01).(4)Changes in cytokines:Interleukin-2 level in the ALRM+CD146+hAD-PCs group was higher than that in the ALRM and CD146+hAD-PCs groups(P<0.05).The interleukin-3 content of the ALRM+CD146+hAD-PCs group was higher than that of the CD146+hAD-PCs group at 2 and 4 weeks(P<0.05).The expression level of granulocyte colony-stimulating factor in the ALRM+CD146+hAD-PCs group was higher than that in the CD146+hAD-PCs group at 1 week,and higher than that in the ALRM group and CD146+hAD-PCs group at 2 weeks(P<0.01).Interferon-γ content in the ALRM group and ALRM+CD146+hAD-PCs group was lower than that in the CD146+hAD-PCs group at 1,2,and 4 weeks of co-culture(P<0.01).(5)Due to the absence of stromal cells in the blank group,UCB CD34+HSPCs could not be counted after 1 week of co-culture and were not subjected to immunophenotyping,colony analysis,or cytokine assays.(6)In summary,ALRM can promote the expansion of UCB CD34+HSPCs in vitro by promoting CD146+hAD-PCs proliferation and cell cycle transformation,which is of great value in hematopoietic stem cell transplantation.

Gene therapy, harnessing the power of CRISPR-Cas9 and/or DNAzyme systems, stands as a pivotal approach in cancer therapy, enabling the meticulous manipulation of genes pivotal to tumorigenesis and immunity. However, the pursuit of precise gene therapy encounters formidable hurdles. Herein, a near-infrared upconversion theranostic nanomachine is devised and tailors for CRISPR-Cas9/DNAzyme systems mediate precise gene therapy. An ingenious logic DNAzyme system consists of Chain 1 (C1)/Chain 2 (C2) and endogenous lncRNA is designed. We employ manganese modified upconversion nanoparticles for carrying ultraviolet-responsive C1-PC linker-C2 (C₂P) chain and Cas9 ribonucleoprotein (RNP), with outermost coats with hyaluronic acid. Upon reaching tumor microenvironment (TME), the released Mn²⁺ ions orchestrate a trifecta: facilitating endosomal escape, activating cGAS-STING signaling, and enabling T1-magnetic resonance imaging. Under near-infrared irradiation, Cas9 RNP/C₂P complex dissociates, releasing Cas9 RNP into the nucleus to perform gene editing of Ptpn2, while C1/C2 chains self-assemble with endogenous lncRNA to form a functional DNAzyme system, targeting PD-L1 mRNA for gene silencing. This strategy remodels the TME by activating cGAS-STING signaling and dual immune checkpoints blockade, thus realizing tumor elimination. Our theranostic nanomachine armed with the CRISPR-Cas9/DNAzyme logic systems, represents a resourceful and promising strategy for advancing cancer systemic immunotherapy and precise gene therapy.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

ObjectiveTo investigate the efficacy of Bushen Shengxue prescription and Yiqi Yangxue prescription in the treatment of chronic aplastic anemia and the effect on T cell subsets and the expression of T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA3）. MethodA total of 585 patients with chronic aplastic anemia who were treated in 19 hospitals in China from May 2018 to June 2021 were enrolled. With the prospective， double-blind and randomized control methods， the patients were randomized into three groups： kidney deficiency group， Qi and blood deficiency group， and control group. The three groups were respectively treated with Bushen Shengxue prescription granule， Yiqi Yangxue prescription granule， and Placebo （half the dose of Bushen Shengxue formula granules）. In addition， all of them were given oral cyclosporin and androgen. The treatment lasted 6 months， with 3 months as a course. The blood routine indexes， T cell subsets， and fusion genes T-bet and GATA3 before and after treatment were analyzed， and the safety indexes were monitored. ResultDuring the observation， a total of 75 cases dropped out and 18 were rejected. Finally， 161 cases in the kidney deficiency group， 164 in the Qi and blood deficiency group， and 167 in the control group were included. After 6 months of treatment， the total effective rate was 98.8% （159/161） in the kidney deficiency group， which was higher than the 79.9% （131/164） in the Qi and blood deficiency group （χ²=30.135， P<0.01） and the 61.7% （103/167） in the control group （χ²=70.126， P<0.01）. The total effective rate was higher in the Qi and blood deficiency group than in the control group （χ²=13.232， P<0.01）. After treatment， the hemoglobin （HGB） content increased significantly in three groups （P<0.05） as compared with that before treatment， particularly the kidney deficiency group （P<0.01）. After treatment， the white blood cell （WBC） count and platelet （PLT） count in the kidney deficiency group and the control group increased compared with those in the Qi and blood deficiency group （P<0.01）. There was no specific difference in neutrophils （ANC） after treatment among the three groups. At the same time point， the level of T helper type 1 （Th1） cells， Th1/Th2 ratio （P<0.05）， level of CD4⁺， and CD4⁺/CD8⁺ ratio （P<0.05） were significantly low in the kidney deficiency group among three groups. There was no significant difference in CD19^-， HLA/DR⁺， and CD25⁺ between the kidney deficiency group and the other two groups， but the T-bet of the kidney deficiency group and the control group was lower than that of the Qi and blood deficiency group （P<0.05）. ConclusionBushen Shengxue prescription exerts therapeutic effect on the aplastic anemia by improving the immunoregulatory mechanism， inhibiting the activity of immune system， modulating T cell subsets， suppressing Th1 and CD4⁺， and promoting bone marrow hematopoiesis. Moreover， it is safe with little side effects， which is worthy of further promotion.

Objective To evaluate the application value of the micro movement sensitive mattress sleep monitoring system(MSMSMS) in the diagnosis of children with obstructive sleep apnea syndrome (OSAS).Methods One hundred and twenty-nine children aged from 3 to 14 years who visited the sleep center of Beijing Children's Hospital Affiliated to Capital Medical University from June 2013 to June 2015 due to sleep snoring were enrolled.Children with acute respiratory infection,cranial facial abnormalities,chronic lung diseases and neuromuscular diseases were excluded.According to the criteria,36 children were diagnosed as OSAS with average age of (7.3 ± 2.5) years,including 28 males and 8 females.Ninety-three non-OSAS children were recruited with average age of (6.3 ± 2.3) years,including 61 males and 32 females.Subjects were monitored with polysomnography(PSG) and MSMSMS simultaneously.Apnea/hypopnea index (AHI) ＞ 5 or obstructive apnea index (OAI) ＞ 1 were used to define whether OSAS existed.The consistency between MSMSMS and PSG in the diagnosis of OSAS and the determination of sleep efficiency were compared.Results The Kappa consistency coefficient of MSMSMS and PSG in the diagnosis of OSAS was 0.70(95％ CI:0.57-0.84),Z =7.99,P ＜ 0.000 1,which indicated the consistency between PSG and MSMSMS was good.The consistency of sleep efficiency of MSMSMS and PSG were compared.Bland-Altman results showed that there were 3％ (5/129 cases)points out of 95％ consistency bound and the interclass correlation coefficient (ICC) was 0.69 which indicated the consistency of 2 methods was good in determination of sleep efficiency.MSMSMS was able to detect respiratory event that was associated with sub-cortical arousals with no electroencephalogram arousal or blood oxygen reduction.Conclusions There is an adequate consistency between MSMSMS and PSG in the diagnosis of children with OSAS and determination of sleep efficiency.The MSMSMS has an advantage in detection of sub-cortical arousals and respiratory event.

Objective To evaluate the effectiveness and safety of mouse nerve growth factor in treating dysphagia in patients with nasopharyngeal carcinoma after radiotherapy.Methods Fifty-eight post-radiotherapy nasopharyngeal carcinoma patients with dysphagia were randomly divided into an observation group and a control group.Both groups received routine treatment,but the observation group was additionally injected with mouse nerve growth factor intramuscularly every day for four weeks.Before and after the 4 weeks of treatment,both groups were evaluated using Kubota's water drinking test,videofluoroscopy and the brief version of the WHO's Quality of Life scale.Results After 4 weeks,the patients in the observation group displayed significantly greater improvement in swallowing compared with the control group.There was a significant difference in the groups' average scores on the drinking water test and in the videofluoroscopy results.Moreover,the patients in the observation group had significantly higher quality of life scores than those in the control group,on average.Conclusions Mouse nerve growth factor may have a rapid and safe therapeutic effect on dysphagia induced by radiation.No obvious adverse reactions were observed.

Objective To investigate the application of non-invasive ventilation in children with airway obstructive diseases,especially those who had obstructive sleep apnea syndrome(OSAS).Methods A case follow-up study was conducted between October 2005 and October 2013 in children who had airway obstruction that led to OSAS or chronic respiratory failure and had been given non-invasive ventilation therapy.Children received non-invasive ventilation support,and pressure titration was performed manually in the sleep center while the mode was chosen according to their disease condition.Pulse rate,oxygen saturation or polysomnography were monitored during the treatment.Some patients went on receiving ventilation support when discharged home depending on their disease status.Patients were followed up every 3,6,or 12 months.Results Thirty-seven patients received non-invasive ventilation treatment till October 2013.Thirty-two cases were boys,and 5 cases were girls.The age ranged from 1 year old and 2 months to 12 years old and 6 months.The underlying diseases included OSAS with adenotonsillar hypertrophy,OSAS with mucopolysaccharidosis,mental retardation,cerebral palsy,morbid obesity,and bronchiolitis obliterans.All the OSAS patients had their snoring and apneas relieved,and respiratory distress and daytime symptoms were improved.Regarding the sleep study parameter,the apnea hypopnea index (P ＜ 0.001),obstructive apnea index (P =0.001),oxygen desaturation index(P =0.001),minimum oxygen saturation (P ＜ 0.001) were improved.Till the end of the study,18children (49％)were still receiving non-invasive ventilation,9 children (24％)stopped ventilation after discharge home,4 children (11％)ceased treatment as their symptoms disappeared and polysomnography data was normal,4 children (11％) lost follow-up 3 months after treatment,and 2 children (5％) died of underlying disease.Conclusions Some children with airway obstruction need non-invasive ventilation support.Non-invasive ventilation therapy can be successfully performed in pediatric population.

Objective To investigate the role of individualized psychological intervention in gastroscopy.Methods 286 patients first underwent gastroscopy were randomly divided into the intervention and control groups,143 patients in each group. Individualized psychological intervention were provided for intervention group,and the patients of control group were just given routine nursing care. All the patients were measured by self rating anxiety scale (SAS) ,which were as measuring standards before and after intervention. At the same time,the blood pressure and heart rate were compared before and after intervention. Visual analogue scale (VAS) was observed to investigate the satisfaction of patients after operation. Results Compared with control group,anxiety rate and degree,blood pressure and heart rate in intervention group were lower( P ＜ 0. 05 ). The test of VAS revealed that the satisfaction of patients in intervention group was significantly higher than that in control group ( P ＜ 0. 05). Conclusion Individualized psychological intervention could improve negative emotion and relieve anxiety and affliction of patients underwent gastroscopy, and the satisfaction degree for gastroscopy was improved.

Objective To assess the value of multi-slice spiral CTA in the diagnosis of iliac vein compression syndrome(IVCS)and secondary thrombosis.Methods The CTA data of 80 controls and 31 patients with IVCS conformed by DSA were retrospectively studied.The inner diameters of the compressed iliac vein in two groups were measured and compression ratios were calculated.The data were compared using t test and Wilcoxon test.The findings of CTA were compared with that of DSA and the accuracy of CTA was analyzed.Results For the control group,the inner diameters of left iliac vein in female group(7.0±2.5)mm were significantly less than that in male group(8.1±2.5)mm(t=2.42,P＜0.05).For the control group,the inner diameters of left iliac vein was(7.6±2.0)mm,and the compression ratios were 0 to 65.41%(median 27.65%).The compression ratios were no statistical differences between male group(0 to 61.36%,median 26.82%)and female group(0 to 65.41%,median 28.75%)(Z=-0.59,P＞0.05).For the patients group,the inner diameters of compression iliac vein was(2.7±1.1)mm.The compression ratios were 55.18% to 100%(median 76.12%).Both inner diameters and compression ratios were statistically different between the control and patients group(t=12.78,P＜0.05;Z=-8.18,P＜0.05).Fifteen of 31 cases with IVCS were accompanied with deep vein thrombosis.The left iliac veins were compress in 28 cases,while right iliac veins were compressed in 2 cases.The left and right iliac common veins were compressed by the left and right jliac common arteries in one case.The results of CTA were consistent with that of DSA in all cases.Conclusions CTA can not only measure the inner diameters of iliae vein compression and calculate compression ratios,but also demonstrate secondary thrombosis clearly.CTA is an effective examination in diagnosis of IVCS and secondary thrombosis.

BACKGROUND:Severe acute respiratory syndrome (SARS),an acute infection with a mutant coronavirus,is characterized by acute pulmonary inflammatory injury accompanied by possible multiple organ injuries. Besides somatic symptoms,affective disorder constitutes an obvious neuropsychiatric symptom in SARS patients.OBJECTIVE:To analyze the clinical features of SARS with neuropsychiatric involvement.DESIGN:A retrospective case analysis.SETTING:Department of Neurology,Department of Respiratory Diseases and Department of Radiology of Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS:Totally 325 SARS patients treated in the Second Affiliated Hospital of Sun Yat-sen University and the 8th People's Hospital of Guangzhou from February 2003 to May 2003 were involved,including 173cases who developed neuropsychiatric symptoms.METHODS:The clinical data of 325 of SARS including 173 cases with neuropsychiatric symptoms were retrospectively analyzed.MAIN OUTCOME MEASURES:Clinical neuropsychiatric symptoms,ophthalmoscopic examination,and pathological findings in autopsy.RESULTS: ①The incidence of neuropsychiatric involvement was about 53.2% in SARS patients. The manifestations consisted mostly of headache (67.1%),dizziness(28.9%),conscious-ness disturbance(10.4%),mental disorder(4.6%), local orientational disorder(0.6%),seizures(1.2%),and affective disorders (30.6%);anxiety occurred in 20.2%,depression in 6.4%,suicidal tendency in 1.2%,and phobia in 7.5% of the patients with neuropsychiatric involvement. ②Among the 325 patients,56 were in critical condition of whom 47 had accompanying neuropsychiatric impairment,with significantly greater incidence than that in mild cases (P＜0.01). ③Ophthalmoscopic examination performed in 4 patients showed blurred margin of the optic disc with retinal venous congestion. ④Autopsy in 2 cases identified edema of the perivascular tissue and vascular wall of the small veins with mononuclear cell and lymphocyte infiltration,neuron degeneration and demyelination in focal brain tissues.CONCLUSION:The manifestations of neuropsychiatric involement can vary in association with the severity of SARS,and the typical pathological changes include edema,neuronal demyelination etc.in the brain.