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 evaluate the efficacy and safety of Weiyan Tongluo Granules in treating chronic atrophic gastritis(CAG)and explore its potential mechanisms.METHODS From June 2020 to December 2022,100 CAG patients with spleen defi-ciency and blood stasis syndrome were enrolled and randomly divided into a trial group(n=50)and a control group(n=50)using a random number table.The trial group received Weiyan Tongluo Granules plus a folic acid placebo,while the control group received fo-lic acid tablets plus a traditional Chinese medicine granule placebo.The treatment course for both groups was 24 weeks,with 8 and 10 dropouts in the trial and control groups,respectively.Post-treatment comparisons included OLGA/OLGIM staging reversal rates,low-grade intraepithelial neoplasia regression rate,SSDPRO-CG(Patient-Reported Outcome Scale for Chronic Gastritis in Spleen-Stomach Diseases)scores,TCM syndrome scores,and safety indicators.Serum levels of PG I,PGⅡ,PGR,and G-17 were measured via ELISA before and after treatment.Gastric mucosal p-NF-κB and CDX2 protein expression levels were analyzed by Western blot,while mRNA levels of IL-1β,IL-6,VIL1,and MUC2 were quantified via qPCR.RESULTS After treatment,the trial group showed sig-nificantly higher OLGA and OLGIM stage reversal rates than the control group(P<0.05,P<0.01),though no significant difference was observed in low-grade intraepithelial neoplasia regression.Both groups exhibited significant improvements in physiological domain scores and total SSDPRO-CG scores(P<0.01),with the trial group outperforming the control group in physiological,independence,psychological domains,and total scores(P<0.05,P<0.01).TCM syndrome scores(total and sub-items:gastric distension,pain,poor appetite,bloating)decreased significantly in both groups(P<0.01),while the trial group showed greater reductions in loose stools and dull complexion(P<0.01).After-treatment,the trial group had significantly lower TCM syndrome scores than the control group(P<0.05,P<0.01).Serum PG I,PGⅡ,PGR,G-17,gastric mucosal p-NF-κB,CDX2,and IL-1β,IL-6,VIL1,MUC2 mRNA levels improved significantly in the trial group(P<0.05,P<0.01),while the control group improved only in PGⅡ(P<0.05,P<0.01).The trial group's improvements in these biomarkers surpassed the control group's(P<0.05,P<0.01).No treatment-related adverse events occurred in either group.CONCLUSION Weiyan Tongluo Granules ameliorate gastric mucosal pathology,clinical symptoms,psychological state,and quality of life in CAG patients without significant adverse effects.Its mechanism may involve sup-pressing the NF-κB pathway to reduce IL-1β and IL-6 expression,downregulating CDX2 to inhibit VIL1 and MUC2 transcription,thereby reversing the vicious cycle of inflammation-intestinal metaplasia.

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.

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.

Premature ovarian insufficiency(POI)is a disease that seriously affects the health and life of women,which is also one of the main causes of female infertility.The incidence of POI is increasing in recent years.The diagnostic criteria of POI are not uniform,and different diagnostic criteria have inconsistent thresholds for various indicators.Strict diagnostic criteria are more conducive to early detection and treatment of POI.At present,complementary and alternative therapies such as traditional Chinese medicine,acupuncture and moxibustion are widely used in the treatment of POI.This article reviewed the recent studies on the complementary and alternative therapy of traditional Chinese medicine in the treatment of POI,and finds that the dialectical treatment of traditional Chinese medicine is based on the perspective of traditional Chinese medicine theory and puts forward corresponding treatment plans for different dialectical types.For example,treatment from the kidney,tonifying kidney filling essence,regulating chongren;Treatment from the liver,soothing the liver,regulating qi,nourishing blood and softening the liver;Treatment from the heart and spleen,nourishing Yangming,tonifying the spleen and nourishing the heart.In terms of acupuncture treatment,regulating menstruation and promoting pregnancy and regulating the conception vessel and toning the governor vessel and toning the kidney are often used to treat POI.In addition,moxibustion regulates body function by stimulating the bladder meridian and conception vessel and governor vessel acupoints.Other methods,such as ear acupoint bean-pressing,acupoint thread-embedding and acupoint injection,have also shown potential in the treatment of POI.This article summarizes the role and characteristics of traditional Chinese medicine in the treatment of POI,in order to provide ideas and basis for clinical treatment of POI.

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.

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.