Objective To observe the clinical efficacy of Dan Qi Yishen Prescription in treating diabetic nephropathy(DN)and explore its renal protective mechanism.Methods A total of 150 patients with DN of qi-yin deficiency complicated with blood stasis syndrome who admitted to Sanya Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Chinese Medicine from October 2021 to April 2023 were divided into a control group and an observation group according to the random number table method,with 75 patients in each group.Both groups were given conventional therapies such as dietary treatment,blood glucose control,blood pressure control,lipid-lowering treatment and treatment of complications.Additionally,the control group was given intravenous injection of reduced glutathione,while the observation group was given oral use of the decoction of Dan Qi Yishen Prescription and ionic introduction of Dan Qi Yishen Prescription in renal region.Thirty days constituted one course of treatment,and both groups were treated for three courses.Before and after treatment,the scores of traditional Chinese medicine(TCM)syndrome,and the levels of renal function indicators,renal hemodynamic indicators,coagulation and fibrinolysis indicators,and vascular endothelial function indicators in the two groups were observed.Results(1)After treatment,the scores of TCM symptoms such as fatigue and weakness,palpitations and shortness of breath,dizziness and tinnitus,spontaneous sweating and night sweating,restlessness and insomnia,and thirst with preference for drinks in the two groups were decreased compared with those before treatment(P<0.01),and the decrease in the observation group was significantly superior to that in the control group(P<0.01).(2)After treatment,the levels of renal function indicators such as blood urea nitrogen(BUN),serum creatinine(SCr),collagen Ⅳ(CⅣ),and plasma laminin(LN)of patients in the two groups were improved compared with those before treatment(P<0.01),and the improvement in the observation group was significantly superior to that in the control group(P<0.01).(3)After treatment,the levels of renal hemodynamic indicators such as systolic maximum blood flow velocity(Vsmax)and diastolic minimum blood flow velocity(Vdmin)of the main renal artery(MRA)and interlobar artery(IRA)in the two groups were increased compared with those before treatment(P<0.05 or P<0.01),while the resistance index(RI)and pulsatility index(PI)of MRA and RI of IRA in the two group as well as PI of IRA in the observation group were decreased compared with those before treatment(P<0.05 or P<0.01).The increase of Vsmax and Vdmin of MRA and IRA as well as the decrease of RI and PI of MRA and IRA in the observation group was significantly superior to that in the control group(P<0.05 or P<0.01).(4)After treatment,the levels of coagulation and fibrinolysis indicators such as prothrombin time(PT),activated partial thromboplastin time(APTT),D-dimer(D-D),and fibrinogen(FIB)in the two groups were improved compared with those before treatment(P<0.01),and the improvement in the observation group was significantly superior to that in the control group(P<0.01).(5)The analysis of vascular endothelial function indicators showed that after treatment,the serum vasohibin 1(VASH-1)level in the two groups was increased compared with that before treatment(P<0.01),and the serum vascular endothelial growth factor(VEGF)and endothelin 1(ET-1)levels were decreased compared with those before treatment(P<0.01),and the increase of serum VASH-1 level as well as the decrease of serum VEGF and ET-1 levels in the observation group was significantly superior to that in the control group(P<0.01).Conclusion Dan Qi Yishen Prescription exerts good clinical efficacy in treating patients with DN of qi-yin deficiency complicated with blood stasis syndrome,and it has protective effect on renal function probably by improving the coagulation and fibrinolysis system and vascular endothelial function of the patients.

Objective:To investigate the effects of microRNA-325-3p(miR-325-3p)over-expression on the invasion and migration of colon cancer cells,and to clarify their mechanisms.Methods:The tumor tissue and the corresponding adjacent normal tissue of 25 patients clearly diagnosed with colon cancer were collected.The expression levels of miR-325-3p and relevant evolutionary and lymphoid interest domain containing protein 1(PRELID1)mRNA were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.The expression levels of miR-325-3p and PRELID1 mRNA and the expression levels of PRELID1 protein in human normal colon cells NCM460 and colon cancer cells SW480,HCT116 and HT-29 were detected by RT-qPCR and Western blotting methods.The SW480 cells were divided into control group,NC-mimics group,miR-325-3p mimics group,miR-325-3p mimics+oe-NC group and miR-325-3p mimics+oe-PRELID1 group.The invasion and migration abilities of cells in various groups were detected by Transwell chamber assay and scratch healing assay,respectively.The expression levels of E-Cadherin,N-Cadherin and Vimentin in the epithelial-mesenchymal transition(EMT)of cells in various groups were detected by Western blotting method.The downstream target genes of miR-325-3p were predicted using the Targetscan bioinformatics website,and the targeted regulatory relationship between miR-325-3p and PRELID1 was verified by the dual-luciferase assay.Results:Compared with adjacent normal tissue,the expression level of miR-325-3p in cancer tissue was significantly decreased(P＜0.05),while the expression level of PRELID1 mRNA was significantly increased(P＜0.05).The expression levels of miR-325-3p and PRELID1 mRNA was negatively correlated in colon cancer tissue(r＜0,R2=0.392,P＜0.001).Compared with NCM460 cells,the expression levels of miR-325-3p in SW480,HCT116 and HT-29 cells were significantly decreased(P＜0.05),and the expression levels of PRELID1 mRNA were significantly increased(P＜0.05).Compared with control group and NC-mimics group,the number of invasive SW480 cells in miR-325-3p mimics group was significantly reduced(P＜0.05),the rate of scratch healing was decreased(P＜0.05),and the expression level of E-Cadherin protein was increased,while the expression levels of N-Cadherin and Vimentin proteins were decreased(P＜0.05).The dual-luciferase assay confirmed that PRELID1 was the direct target of miR-325-3p.Compared with miR-325-3p mimics+oe-NC group,the number of invasive SW480 cells in miR-325-3p mimics+oe-PRELID1 group was significantly increased(P＜0.05),the rate of scratch healing was increased(P＜0.05),the expression level of E-Cadherin protein was decreased,and the expression levels of N-Cadherin and Vimentin proteins were increased(P＜0.05).Conclusion:Over-expression of miR-325-3p can inhibit EMT process by down-regulating PRELID1 expression,thereby inhibiting SW480 cell invasion and migration.

Objective To discuss the perioperative care and wound protection of xenotransplantation from genetically edited pigs to monkeys, with the goal of improving the success rate of such experimental procedures. Methods From October 2022 to October 2023, perioperative care and wound protection were performed on 7 recipient rhesus monkeys undergoing xenotransplantation of genetically edited pig tissues and organs. Customized wound protective garments were designed based on monkeys' size and surgical area to protect the wounds, alongside meticulous perioperative care. This included preoperative preparation and medication, intraoperative monitoring of physiological indicators and anesthesia management, and postoperative care comprising wound protection, observation and monitoring, and nutritional support. Results All seven monkeys successfully underwent xenotransplantation. With the aid of protective garments and detailed care, all surgical wounds healed by first intention, and postoperative recovery was satisfactory. Conclusion Proper care and wound protection during xenotransplantation from genetically edited pigs to monkeys not only promote wound healing, but also alleviate pain and harm to animals. This has significant implications for advancing experimental research in pig-monkey xenotransplantation and enhancing animal welfare.

Objective:To investigate the optimal serum antibody titer in acute stage for the diagnosis of Mycoplasma pneumoniae (MP) infection by passive agglutination, and to evaluate the clinical diagnostic value of different antibody titers.Methods:A cross-sectional study.Eighty-eight pairs of clinical serum samples were collected from children with MP infection treated at the Department of Pediatrics in Shengjing Hospital of China Medical University from December 2016 to February 2017 and Children′s Hospital of Baotou in November 2019.The four-fold change of the double serum specific antibody titer was used as the gold standard, and the receiver operating characteristic (ROC) curve was plotted.When detecting the single serum in acute stage, different antibody titers were used as positive criteria to evaluate their clinical application value in the diagnosis of MP infection and find the most appropriate serum antibody titer as the diagnostic cut-off value.Results:(1)When the serum specific antibody titer ≥1∶40 was used as the positive criterion, the sensitivity was 72.9%, the area under the ROC curve was 0.817, and the specificity was 87.5%, which might cause overdiagnosis.When the serum specific antibody titer ≥1∶160 was used as the positive criterion, the specificity was 97.5%, the area under the ROC curve was 0.775, and the sensitivity was 52.1%, which might cause missed diagnosis.When the serum specific antibody titer ≥1∶80 was used as the positive criterion, the sensitivity was 60.4%, the specificity was 97.5%, and the area under the ROC curve was 0.823, overall performing better compared with the said two criteria.(2)After the disease lasted at least 5 days, blood samples were collected.About 72.5% of the children had antibodies, and 60.0% of the children had antibody titers ≥1∶80.Conclusions:(1)When the passive agglutination method is used to detect MP infection, antibody titer ≥1∶80 is recommended as the diagnostic standard.However, in clinical practice, the diagnosis of MP infection depends on clinical and other laboratory test results.(2) It is appropriate to collect blood samples on 5-7 days of illness.If MP infection is clinically suspected, and an antibody titer of 1∶40 is also suggestive, it can perform cooperative diagnosis based on molecular biology lab results or retest at a shorter interval.

Accumulating evidence has demonstrated that apoptotic vesicles(apoVs)derived from mesenchymal stem cells(MSCs;MSC-apoVs)are vital for bone regeneration,and possess superior capabilities compared to MSCs and other extracellular vesicles derived from MSCs(such as exosomes).The osteoinductive effect of MSC-apoVs is attributed to their diverse contents,especially enriched proteins or microRNAs(miRNAs).To optimize their osteoinduction activity,it is necessary to determine the unique cargo profiles of MSC-apoVs.We previously established the protein landscape and identified proteins specific to MSC-apoVs.However,the features and functions of miRNAs enriched in MSC-apoVs are unclear.In this study,we compared MSCs,MSC-apoVs,and MSC-exosomes from two types of MSC.We generated a map of miRNAs specific to MSC-apoVs and identified seven miRNAs specifically enriched in MSC-apoVs compared to MSCs and MSC-exosomes,which we classified as apoV-specific miRNAs.Among these seven specific miRNAs,hsa-miR-4485-3p was the most abundant and stable.Next,we explored its function in apoV-mediated osteoinduction.Unexpectedly,hsa-miR-4485-3p enriched in MSC-apoVs inhibited osteogenesis and promoted adipogenesis by targeting the AKT pathway.Tailored apoVs with downregulated hsa-miR-4485-3p exhibited a greater effect on bone regeneration than control apoVs.Like releasing the brake,we acquired more powerful osteoinductive apoVs.In summary,we identified the miRNA cargos,including miRNAs specific to MSC-apoVs,and generated tailored apoVs with high osteoinduction activity,which is promising in apoV-based therapies for bone regeneration.

Accumulating evidence has demonstrated that apoptotic vesicles(apoVs)derived from mesenchymal stem cells(MSCs;MSC-apoVs)are vital for bone regeneration,and possess superior capabilities compared to MSCs and other extracellular vesicles derived from MSCs(such as exosomes).The osteoinductive effect of MSC-apoVs is attributed to their diverse contents,especially enriched proteins or microRNAs(miRNAs).To optimize their osteoinduction activity,it is necessary to determine the unique cargo profiles of MSC-apoVs.We previously established the protein landscape and identified proteins specific to MSC-apoVs.However,the features and functions of miRNAs enriched in MSC-apoVs are unclear.In this study,we compared MSCs,MSC-apoVs,and MSC-exosomes from two types of MSC.We generated a map of miRNAs specific to MSC-apoVs and identified seven miRNAs specifically enriched in MSC-apoVs compared to MSCs and MSC-exosomes,which we classified as apoV-specific miRNAs.Among these seven specific miRNAs,hsa-miR-4485-3p was the most abundant and stable.Next,we explored its function in apoV-mediated osteoinduction.Unexpectedly,hsa-miR-4485-3p enriched in MSC-apoVs inhibited osteogenesis and promoted adipogenesis by targeting the AKT pathway.Tailored apoVs with downregulated hsa-miR-4485-3p exhibited a greater effect on bone regeneration than control apoVs.Like releasing the brake,we acquired more powerful osteoinductive apoVs.In summary,we identified the miRNA cargos,including miRNAs specific to MSC-apoVs,and generated tailored apoVs with high osteoinduction activity,which is promising in apoV-based therapies for bone regeneration.

Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.

Objective: To compare behavioral and emotional health among first-born children and the only-child in Harbin, as well as associated factors including parents, family background, parenting and family environment. Methods: A questionnaire survey was conducted for 156 parents of first-born children and the only-children, matched in age (<3 months), class and gender. Achenbach Child behavior scale (CBCL), the Self-evaluation of Anxiety Scale (SAS), the Self-rating Depression Scale(SDS), the Parents Rearing Behavior Questionnaire (CRPR) and the Family Assessment Device Scale (FAD) were used. Results: There was no statistically significant difference in the scores of each dimension of children's emotional and behavioral health between the two groups(17.88±5.93)(19.13±6.01),total score(t=-0.74，P>0.05). There was no statistical difference in anxiety and depression between the two groups of parents(χ2=0.51，0.40，P>0.05); In terms of parenting style, the acceptance and encouragement achievement score for first-born children was significantly higher than that of the only child (t=2.10,2.12, P<0.05). In terms of family functions, there was no statistical difference in total function (t=-0.43, P>0.05). Behavioral problems associated with parents' anxiety, depression, parental rearing style and family function. Regression analysis showed that behavioral problems were mainly affected by sibling relationship for first-born children(B=8.74) and family role function for the only child (B=1.27). Conclusion No significant differences in behavioral and emotional health between first-born child and the only child are observed. However, harmonious sibling relationship, emotionally supportive parents and home environment could help improving behavioral and emotional health.

OBJECTIVETo investigate the expression of interleukin-9 (IL-9) in colon cancer tissues and its clinical significance.

METHODSImmunohistochenmistry and qRT-PCR were used to detect the expressions of IL-9 protein and mRNA in 92 colon cancer tissues and paired adjacent normal tissues. The correlation of IL-9 expressions with the clinicopathological features and prognosis of the patients was analyzed.

RESULTSIL-9 protein and mRNA expressions were significantly higher in adjacent normal tissues than in the colon cancer tissues ( < 0.001). In colon cancer patients, IL-9 expression was significantly correlated with TNM stage (=0.013), Ducks stage (=0.025) and lymph node metastasis (=0.004) but not with gender, age, tumor size, differentiation or hepatic metastasis ( > 0.05). The survival time of colon cancer patients with positive IL-9 expression was significantly longer than that of patients negative for IL-9 expression (=0.015).

CONCLUSIONSIL-9 expression is lowered in colon cancer tissues compoved with in the adjacent normal tissues. IL-9 expression is negatively correlated with TNM staging, Ducks staging and lymph node metastasis but positively with good prognosis, suggesting its important role in the tumor microenvironment of colon cancer.

Our previous studies found that mitochondrial uncouplers CCCP and niclosamide inhibited artery constriction and the mechanism involved AMPK activation in vascular smooth muscle cells. BAM15 is a novel type of mitochondrial uncoupler. The aim of the present study is to identify the vasoactivity of BAM15 and characterize the BAM15-induced AMPK activation in vascular smooth muscle cells (A10 cells). BAM15 relaxed phenylephrine (PE)-induced constricted rat mesenteric arteries with intact and denuded endothelium. Pretreatment with BAM15 inhibited PE-induced constriction of rat mesenteric arteries with intact and denuded endothelium. BAM15, CCCP, and niclosamide had the comparable IC value of vasorelaxation in PE-induced constriction of rat mesenteric arteries. BAM15 was less cytotoxic in A10 cells compared with CCCP and niclosamide. BAM15 depolarized mitochondrial membrane potential, induced mitochondrial fission, increased mitochondrial ROS production, and increased mitochondrial oxygen consumption rate in A10 cells. BAM15 potently activated AMPK in A10 cells and the efficacy of BAM15 was stronger than that of CCCP, niclosamide, and AMPK positive activators metformin and AICAR. In conclusion, BAM15 activates AMPK in vascular smooth muscle cells with higher potency than that of CCCP, niclosamide and the known AMPK activators metformin and AICAR. The present work indicates that BAM15 is a potent AMPK activator.