Objective To explore the application effect of bedside critical ultrasound in volume management of acute kidney injury(AKI)donors.Methods Clinical data of 56 AKI donors and 106 recipients from the Transplantation Center of People's Hospital of Guangxi Zhuang Autonomous Region from October 1,2020 to May 31,2022 were collected.They were divided into the critical ultrasound group(34 donors,66 recipients)and the traditional central venous pressure(CVP)group(22 donors,40 recipients)according to the volume management methods.The AKI stage and recovery time,renal function indicators(serum creatinine(Scr),cystatin C,estimated glomerular filtration rate),donor Remuzzi score,cold ischemia time,biventricular inotrope usage rate and time,delayed graft function(DGF)incidence and recovery time,and renal function indicators at 6 and 12 months after surgery were compared and analyzed between the two groups.Results There were no statistically significant differences in gender,age,body weight,AKI stage,pre-acquisition renal function indicators,biventricular inotrope usage rate,renal function indicators at 6 and 12 months after surgery,DGF recovery time,donor Remuzzi score and cold ischemia time of the donors and recipients between two groups(all P>0.05).The AKI recovery time,continuous renal replacement therapy rate and biventricular inotrope usage time of donors in the critical ultrasound group were shorter or lower than those in the traditional CVP group(all P<0.05).The incidence of DGF in recipients of the critical ultrasound group was lower than that of the traditional CVP group(P<0.05).Subgroup analysis showed that there was no statistically significant difference in Scr at 6 and 12 months after surgery in recipients of the critical ultrasound group(P>0.05),while the Scr at 12 months after surgery was higher than that at 6 months in recipients of the traditional CVP group(P<0.05).Conclusions AKI kidneys may be used for kidney transplantation after active maintenance.Bedside critical ultrasound has unique advantages in volume management of AKI donors and may improve the function of AKI kidneys to a certain extent.

Objective To explore the biological functions and mechanisms of PIWI-interacting RNA(piRNA)in cervical cancer(CC).Methods RT-qPCR was used to detect the expression level of piRNA-2732 in CC tissue,CaSki cells and End1/E6E7 cells.EpiQuik m6A RNA methylation quantification kit was used to detect the methylation level of m6A RNA in CaSki cells.The expression levels of methyltransferases(METTL3,METTL14 and WTAP)and demethylases(FTO,ALKBH5)mRNA in CaSki cells were detected by RT-qPCR.After culturing CaSki cells to logarithmic growth stage,they were divided into six groups:piRNA-2732 mimic negative control group(mi-NC group),piRNA-2732 mimic group(mi-2732 group),piRNA-2732 inhibitor negative control group(in-NC group),piRNA-2732 inhibitor group(in-2732 group),piRNA-2732 mimic+METTL3 knockdown control group(mi-2732+si-NC group),and piRNA-2732 mimic+METTL3 knockdown group(mi-2732+si-METTL3 group).The viability of CaSki cells was detected by CCK8 assay.Colony formation assay was used to detect the proliferation ability of CaSki cells.Transwell experiment was used to detect the migration and invasion ability of CaSki cells.RT-qPCR and Western blot were used to detect the expression of methyltransferase like protein 3(METTL3).Transfected METTL3 wild-type(METTL3-WT)and METTL3 mutant(METTL3-MUT)in the mi-NC group,mi-2732 group,in-NC group,and in-2732 group respectively,and detected the effect of piRNA-2732 on METTL3 through dual luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of piRNA-2732(3.84±1.08 vs 1.32±0.53)was significantly higher in the cancer tissues of CC patients,and the difference was statistically significant(t=5.115,P<0.001).Compared with end1/E6E7 cells,the expression of piRNA-2732(1.00±0.13 vs 1.67±0.16)in CaSki cells was significantly higher,and the difference was statistically significant(t=5.632,P<0.01).Compared with mi-NC group,mi-2732 group promoted the viability,proliferation,migration and invasion of CaSki cells,and the differences were statistically significant(t=4.410～11.040,all P<0.01).Compared with mi-NC group,mi-2732 group increased m6A RNA methylation level and METTL3 mRNA and protein,the differences were statistically significant(t=6.176,9.211,12.550,all P<0.05).The results of dual luciferase reporter gene testing showed that compared with the mi-NC+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-WT group was significantly increased(t=11.850).Compared with mi-2732+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-MUT group was significantly lower(t=12.740),and the difference was statistically significant(all P<0.000 1).Compared with in NC+METTL3-WT group,the relative luciferase activity of in-2732+METTL3-WT group was significantly lower(t=7.828),compared with in-2732+METTL3-WT group,the relative luciferase activity of CaSki cells in in-2732+METTL3-MUT group was significantly increased(t=8.146),and the difference was statistically significant(all P<0.001).Compared with mi-2732+si-NC group,the expression level of m6A in mi-2732+si-METTL3 group was significantly lower,and the difference was statistically significant(t=7.630,P<0.01).Compared with mi-2732+si-NC group,the proliferation ability,colony number,cell migration and invasion ability of CaSki cells in mi-2732+si-METTL3 group were significantly decreased,and the differences were statistically significant(t=3.695～4.891,all P<0.001).Conclusion piRNA-2732 is overexpressed in CC tissues and cells,and piRNA-2732 promotes tumor development in CC through METTL3 mediated m6A methylation.

Objective:To explore the effect of EAERI (E: environment; A: accompany; E: emotion; R: respect; I: information) support strategies in the application of nurse second victims in restoring the passage of a patient safety incident, and to provide support to nurse second victims experiencing a safety incident in restoring the incident and mitigating secondary victimization from the incident.Methods:The method of historical comparison was adopted. A convenience sampling method was used to select nurses who experienced patient safety events in Shanxi Bethune Hospital (Shanxi Academy of Medical Sciences) from January 2023 to February 2024 as the study subjects, which were divided into observation group and control group by the historical control grouping method. The observation group used the EAERI support strategy for patient safety incident reduction, and the control group used the conventional reduction method. The second victims of the nurses in both groups were assessed for their level of harm and support in the patient safety incident using the Second Victim Experience and Support Tool before and after the event reduction, respectively.Results:A total of 124 second victims completed the study. There were 63 cases in the control group, including 13 males and 50 females, with an age of (30.24 ± 4.91) years old. There were 20 males and 41 females in the observation group, with an age of (30.85 ± 5.50) years old. Before the restoration event, there was no significant differences in the total scores of victim experience and support, the degree of injury and the degree of support between the two groups (all P>0.05). The total scores of victim experience and support, the degree of injury and the degree of support were (89.35 ± 10.69), (44.57 ± 6.19), (44.78 ± 9.40) points in the control group, and (64.13 ± 9.36), (33.56 ± 8.84), (30.56 ± 3.17) points in the observation group. The differences were significant between the two groups ( t=14.02, 8.07, 11.29, all P<0.01). Conclusions:The EAERI support strategy is feasible and effective. The implementation of the EAERI support strategy can reduce the degree of injury when the second victim of the nurse restores the safety incident and improve the degree of support.

Recent rapid developments in molecular biological techniques have allowed the use of gene editing,as a means of genome modification,for the establishment of experimental animal models,with high efficiency,accuracy,and flexibility.This article mainly summarizes the construction and application of the latest gene-editing techniques in animal models,including pigs,non-human primates and dogs.It provides a theoretical reference for the application and in-depth study of gene-editing techniques in large experimental animals,which may better simulate human diseases,and for further studies of the potential pathogenesis of biomedical and human complex diseases.

Objective:To observe the clinical efficacy of Tuina(Chinese therapeutic massage)plus Liu Zi Jue for adenoid hypertrophy(AH)due to Qi deficiency of lung and spleen and its effects on the adenoid-nasopharyngeal cavity ratio(A/N value)and traditional Chinese medicine(TCM)constitution scale score.Methods:A total of 64 children with AH due to Qi deficiency of the lung and spleen were randomized into a trial group and a control group,with 32 cases in each group.The trial group was treated with Tuina plus Liu Zi Jue,and the control group was treated with Tuina alone.The Tuina manipulations were the same in the two groups.Tuina treatment was performed 3 times a week,with an interval of at least 1 d.Liu Zi Jue was performed 2 sets each time and 5 times a week.After 8 weeks of treatment and at the 2-month follow-up,the clinical efficacy was compared between the two groups,and the changes in the main clinical symptom score,A/N value,and TCM constitution scale score were observed.Results:After treatment,the total effective rate of the trial group and the control group was 96.9%and 93.8%,respectively.The difference between the two groups was not statistically significant(P>0.05).At the follow-up,the total effective rate of the trial group was 93.8%,higher than 90.6%in the control group(P<0.05).After treatment and at the follow-up,the main clinical symptom score in both groups was lower than that before treatment(P<0.05);at the follow-up,the main clinical symptom score in the trial group was lower than that in the control group(P<0.05).After treatment,the A/N value in both groups was lower than that before treatment(P<0.05),and the value in the trial group was lower than that in the control group;however,the difference between the two groups was not statistically significant(P>0.05).After treatment and at the follow-up,the TCM constitution scale score in both groups was lower than that before treatment(P<0.05);the score after treatment in the trial group was lower than that in the control group(P<0.05).Conclusion:Tuina plus Liu Zi Jue can effectively improve the main clinical symptoms in children with AH due to Qi deficiency of lung and spleen,reduce the A/N value and TCM constitution scale score,with stable efficacy.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

Diabetic kidney disease(DKD)is one of the common microvascular complications in diabetes mellitus patients,with oedema,proteinuria,and continuous decline in renal function as the main clinical manifestations,and traditional Chinese medicine(TCM)believes that diabetic kidney disease is caused by a deficiency of Qi and Yin,which runs through the whole process of the disease.In recent years,academician Tong Xiaolin's theory of"state-target theory"provides new ideas for the treatment of diabetic nephropathy.The"state"is a general term for the overall manifestation of the patient's condition.The overall manifestation of the condition of diabetic kidney disease is a"state of deficiency and impairment"with Qi and Yin deficiency as the key mechanism of the disease.The"target"refers to the specific symptoms of the disease or abnormal examination results,which are the clinical symptoms and abnormal indicators caused by diabetic kidney disease.Ginseng astragali dihuang soup is effective in the treatment of diabetic nephropathy,and the application of this formula based on the theory of state-target can make the therapeutic effect more significant,achieve the therapeutic effect of supporting positive Qi,replenishing the deficiency and benefiting the loss,and improve the clinical symptoms and the abnormal physicochemical indexes of diabetic nephropathy,and the state-target co-adjustment,which can provide a new way of thinking for the diagnosis and treatment of diabetic nephropathy for the clinicians.

Objective To investigate the clinical efficacy of the traditional Chinese medicine(TCM)throat opening and brightening method in treating unilateral vocal cord paralysis(UVCP).Methods Sixty patients with UVCP were prospectively collected and randomly assigned to two groups:the Chinese herbal medicine group(trea-ted with Buyang Huanwu Decoction,n=30)and the throat opening and brightening method group(treated with TCM throat opening and brightening method,n=30).The clinical studies that had utilized injection laryngoplasty for the treatment of UVCP(historical control group).Evaluation indicators included the voice handicap index-10(VHI-10),GRBAS-G,objective acoustic measurements of voice(vocal intensity,F0,shimmer,jitter,HNR),and aerodynamic measurements(maximum phonation time,MPT).Results Before treatment,no significant differences were observed between the two groups in all the evaluation indicators(P＞0.05).Post-treatment,the throat open-ing and brightening method group demonstrated significant improvements in VHI-10,GRBAS-G,shimmer,jitter,HNR,and MPT compared to pre-treatment values(P＜0.01),and these improvements were superior to those in the Chinese herbal medicine group.Pre-treatment VHI-10,GRBAS-G,and shimmer scores in the throat opening and brightening method group were significantly higher than those in the historical literature group(P＜0.01).Af-ter treatment,no significant differences were noted in any assessed parameters between the two groups(P＞0.05).Conclusion The TCM throat opening and brightening method significantly enhances phonatory function and quality of life in patients with UVCP,showing comparable efficacy to injection laryngoplasty.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

Recent rapid developments in molecular biological techniques have allowed the use of gene editing,as a means of genome modification,for the establishment of experimental animal models,with high efficiency,accuracy,and flexibility.This article mainly summarizes the construction and application of the latest gene-editing techniques in animal models,including pigs,non-human primates and dogs.It provides a theoretical reference for the application and in-depth study of gene-editing techniques in large experimental animals,which may better simulate human diseases,and for further studies of the potential pathogenesis of biomedical and human complex diseases.