Objective To explore the analgesic initiation mechanism of"three-manipulations and three-acupoints"of tuina on minor chronic constriction injury(minor CCI)model rats.Methods According to the random number table method,35 SD rats were randomly divided into five groups:normal group,sham group,model group,tuina group,and tuina+MK-801 group.The model group,tuina group,and tuina+MK-801 group were subjected to ligation of the right sciatic nerve trunk to establish a minor CCI rat model.The sham group was only exposed to the right sciatic nerve without ligation,and the normal group was not subjected to any operation.The normal group was not subjected to any intervention measures.On the seventh day after modeling,the model group and the sham group underwent 9 minutes of grasping restraint,while the tuina group underwent one intervention of three-manipulations(point method,dialing method,and kneading method)and three-acupoints(right"Yinmen"(BL37),"Chengshan"(BL57),and"Yanglingquan"(GB34)acupoints)with each manipulation and acupoint intervention for 1 minute for a total of 9 minutes.The tuina+MK-801 group received intrathecal injection of MK-801 from the fifth to seventh days after modeling,with a dose of 6 μg(10 μL)per day,tuina intervention was performed 30 minutes after the last intrathecal injection,and the specific operation of tuina was the same as that of the tuina group.Before modeling,after modeling,and after intervention,each group of rats was subjected to cold sensitivity threshold(CST)and mechanical withdrawal threshold(MWT)testing.After intervention,immunohistochemistry was used to detect the positive expression of cyclic guanosine monophosphate(cGMP)in the spinal dorsal horn(SDH)at L4-6 segments;protein expressions of N-methyl-D-aspartate receptor 1(NMDAR1),neurogenic nitric oxide synthase(nNOS),soluble guanylyl cyclase β(sGCβ),and protein kinase G1(PKG1)in SDH at L4-6 segments were detected by Western blotting;mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 in SDH at L4-6 segments were detected by real-time PCR.Results Compared with the normal and sham groups,after modeling,CST increased and MWT decreased in the model group,tuina group and tuina+MK-801 group(P＜0.05);after intervention,the positive protein expression of cGMP was increased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were increased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were increased in SDH at L4-6 segments in the model group(P＜0.05).Compared with the model group,after intervention,CST decreased and MWT increased in the tuina group and tuina+MK-801 group(P＜0.05);the positive protein expression of cGMP was decreased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were decreased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were decreased in SDH at L4-6 segments in the tuina group and tuina+MK-801 group(P＜0.05).Conclusion One-time tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia induced by peripheral nerve injury,which may initiate analgesia through the NMDAR1/cGMP/protein kinase G signaling pathway,thereby exerting immediate analgesic effect.

Objective:To investigate the characteristics of early motor development in small for gestational age (SGA) infants at high risk of brain injury.Methods:This study retrospectively enrolled a total of 81 SGA infants and appropriate for gestational age (AGA) infants who were at high risk of brain injury and attended outpatient follow-up visits in Xi'an Children's Hospital from February to October 2022. Seventeen SGA infants (SGA group) and 24 AGA infants (AGA group) were assessed for motor development using the Test of Infant Motor Performance (TIMP) at 2-5 weeks of corrected age (CA) and 20 SGA infants (SGA group) and 20 AGA infants (AGA group) were assessed at 14-17 weeks of CA. Independent samples t-test, rank-sum test, and Chi-square test were used to compare the demographic characteristics, high-risk factors of brain injury, and TIMP scores between the two groups. Results:At 2-5 weeks and 14-17 weeks of CA, the birth weights of SGA group were both less than those of AGA group [(1 817.1±440.3) vs. (2 630.0±560.9) g, t=－4.98; (1 752.0±434.4) vs. (2 226.3±699.8) g, t=－2.58; both P<0.05], but there were no significant differences in gestational age at birth or high-risk factors of brain injury between the two groups (all P>0.05). (1) At 2-5 weeks of CA: SGA group had lower total TIMP score [(71.6±13.7) vs. (80.5±11.5) scores, t=－2.26, P=0.029], elicited item score [61.0 scores (41.0-85.0 scores) vs. 69.1 scores (49.0-96.0 scores), Z=－2.15, P=0.037], sitting position score [8.8 scores (3.0-19.0 scores) vs. 11.2 scores (5.0-22.0 scores), Z=－2.07, P=0.038], and prone position score [(9.8±3.1) vs. (12.3±3.1) scores, t=－2.19, P=0.034] when compared with AGA group. (2) At 14-17 weeks of CA: The standing position score of the SGA group was lower than that of the AGA group [6.5 scores (4.0-11.0 scores) vs. 7.7 scores (2.0-11.0 scores), Z=－2.05, P=0.040], but no statistical difference was observed in the total TIMP score or the scores of sitting, supine, prone, turning, and lateral positions between the two groups (all P>0.05). Conclusion:Early motor performance of SGA infants is inferior to AGA infants before five months of age, which is embodied in the poor head control at 2-5 weeks of CA that further affects the stability of standing posture in them at 14-17 weeks of CA.

Objective: To investigate the impact of tumor necrosis factor ligand superfamily member 13(TNFSF13) derived from M2 macrophages on temozolomide(TMZ) resistanceviaregulating interferon regulatory factor 8(IRF8) in glioblastoma(GBM) cells. Methods: Immunohistochemistry(IHC) was used to detect the expression of TNFSF13 in normal brain tissues and GBM tissues. ELISA was used to measure the expression of TNFSF13 in the conditioned media(CM) of M0-type macrophages and M2-type macrophages. M0-CM and M2-CM were used to culture U251 sensitive(U251/S) and resistant(U251/R) cells. The TMZ treatment group was also treated with 800 μmol/L TMZ. The U251/R cells were divided into the following groups: con group, M2vector-CM group, M2vector-CM+TMZ group, M2TNFSF13-CM group, M2TNFSF13-CM+TMZ group, si-IRF8 group, and si-IRF8+M2TNFSF13-CM group. CCK-8 assay was used to detect cell viability and calculate the IC50value. Transwell assay was used to detect cell invasion. Flow cytometry was used to detect apoptosis. Western blot was used to detect the expression of IRF8. Nude mouse xenograft models were constructed and the nude mice were divided into the following groups: U251+M2si-NCgroup, U251+M2si-TNFSF13group, U251+M2si-NC+TMZ group, U251+M2si-TNFSF13+TMZ group. The tumor volume and mass of each group were measured, and IHC was used to detect the expression of TNFSF13 and CD206 in tumor tissues of each group. Results: Compared with adjacent tissues and M0-CM, the expression of TNFSF13 was up-regulated in cancer tissues and M2-CM. Compared with the M0-CM group, the IC50value of TMZ and the number of cell invasions in U251/S and U251/R cells in the M2-CM group significantly increased(allP<0.05). Overexpression of TNFSF13 in M2 macrophages could promote the IC50value of TMZ in U251/R cells, promote cell invasion, and inhibit cell apoptosis(allP<0.05). Overexpression of TNFSF13 promoted the expression of IRF8, and knocking down IRF8 could attenuate the TMZ resistance of U251/R mediated by overexpression of TNFSF13.In vivostudies showed that knocking down TNFSF13 alone or combined with TMZ treatment significantly inhibited tumor growth and reduced the expression of TNFSF13 and CD206. Conclusion TNFSF13 derived from M2 macrophages promotes TMZ resistance in GBM cells by activating IRF8.

Objective:To investigate the effect of gracilis flap in repair of radiation-induced vesicovaginal fistula.Methods:The data of 18 patients with radiation-induced vesicovaginal fistula treated in the Sixth People’s Hospital Affiliated to Shanghai Jiaotong University School of Medicine from March 2021 to August 2022 were retrospectively reviewed. Their age was (57.3±10.4) years. All patients underwent radical surgery for cervical cancer, and received (24.6±2.8)(range from 20 to 30)times of radiotherapy after surger. The median time between the end of radiotherapy and the onset of vesicovaginal fistula was 14.0(7.8, 18.2)months. The median duration of fistula urine was 12.0(9.8, 18.0)months. All patients were required to use median 19.5(15.8, 27.5) pads per day before surgery. The life quality score(QOL)of 18 cases was median 5.0(5.0, 6.0) points. Three cases had performed laparoscopic vesicovaginal fistula repair, two cases had underwent transvaginal vesicovaginal fistula repair, one case had underwent transvaginal and laparoscopic vesicovaginal fistula repair successively, and the remaining 12 cases were new vesicovaginal fistulas. Two cases were combined with rectovaginal fistulas. All patients underwent the repair of vesicovaginal fistula with gracilis flap interposition in prone and folded knife position, by transvaginal route, the vesicovaginal fistula was mobilized and the two layers were closed, and the vascular pedicle gracilis flap of left inner leg was romoved under the skin tunnel to repair the vesicovaginal fistula. Meanwhile, two cases combined rectovaginal fistulas were repaired and closed the rectovaginal fistulas. The urinary catheters were removed at 3 weeks after the operation and urination was recorded.Results:All patients underwent smooth surgery in (96.6±13.2) minutes. The median follow-up was 13.0(9.8, 20.2)(range from 6 to 24)months. The median number of urine pads used per day in 18 patients was 2.0(1.0, 11.8), and significantly reduced ( P<0.01).QOL score was median 1.0(0, 4.2) point and significantly reduced ( P<0.01).Successful outcome was achieved in 12 patients with no leakage of urine in the vagina. Two cases developed urinary incontinence and required conservative treatment, but the curative effect was poor. Two cases still had vaginal urine leakage performed vesicovaginal fistula repair again. One case was successfully repaired without significant urine leakage.The other case still had significant urine leakage and the QOL score was 3 points. She refused further treatment for self-satisfied. Two cases still had vesicovaginal fistula and rectovaginal fistula after the surgery, and refused further surgery. Conclusions:Repair with gracilis flap interposition is a surgical method with few complications and reliable surgical effect for patients with radiation-induced vesicovaginal fistula.

Objective To summarize the occurrence of complications in renal graft biopsy,and to analyze the indications for puncture and types of pathological diagnosis.Methods The data of 703 samples of ultrasound-guided renal graft biopsy from 644 kidney transplant recipients from January 1,2017,to December 31,2022 was retrospectively analyzed.The puncture qualification rate,complications,indicative biopsy indications and pathological diagnosis types were analyzed.The application of surveillance biopsy and pathological diagnosis were also analyzed.Results The qualification rate of renal tissue puncture biopsy was 99.9％,and the complications of puncture bleeding included one sample of perinephric hematoma and one sample of hematuria.Increased serum creatinine(76.8％)and proteinuria(13.8％)were the main indications for puncture,and 48 samples(6.8％)were surveillance biopsy for the assessment of therapeutic effects.A total of 399 samples of pathological diagnosis of rejection,including 293 samples of cellular rejection reaction,60 samples of antibody rejection reaction,and 46 samples of mixed rejection reaction.One hundred and ninety-five samples of recurrence or new-onset kidney disease,mainly including 144 samples of IgA nephropathy and 42 samples of focal segmental glomerulosclerosis.Fifty-seven samples of infection related kidney disease,including 56 samples of BK virus-associated nephropathy(BKVAN).Thirty-one samples of calcineurin inhibitor(CN1)nephrotoxicity injury,including 15 samples of acute CNI nephrotoxicity injury and 16 samples of chronic CNI nephrotoxicity injury.Forty-five samples for other diagnoses.Conclusions The success rate and safety of renal graft biopsy are high,and at present,cellular rejection reaction is still the main pathological diagnosis of indicative biopsy for renal graft.

Objective By observing the effects of"three methods and three points"tuina technique on the expression of interleukin-17F(IL-17F),interleukin-17 receptor C(IL-17RC),activator 1 of nuclear transcription factor-κB(Act1),tumour necrosis factor receptor-associated factor 6(TRAF6)and M1 microglial cell expression in the spinal dorsal horn of rats with mild chronic compressive injury(minor CCI)model,we explored the immediate analgesic mechanism of tuina on peripheral neuropathic pain(pNP).Methods Thirty-six SD rats were divided into the sham group,the model group and the tuina group according to the random number method,twelve rats in each group,and the minor CCI model was replicated by ligating the right sciatic nerve.The rats in the tuina group were subjected to pointing,plucking and kneading at the BL37,BL57 and GB34 points on the affected side using a tuina simulator,while the sham group and the model group were only grasped and restrained,and were intervened for one time.The mechanical pain test and cold plate test were used to evaluate the response of rats to mechanical stimulation and cold stimulation after immediate intervention.The protein expression of IL-17F and TRAF6 in the spinal dorsal horn of rats in each group was detected by Western blotting.The mRNA expression of IL-17F,IL-17RC,Act1 and TRAF6 in the spinal dorsal horn of rats in each group was detected by real-time PCR.The average fluorescence intensity of M1 microglia in the spinal dorsal horn of rats in each group was detected by immunofluorescence.Results Behavioral results showed that before intervention,compared with the sham group,paw mechanical withdraw threshold(PMWT)decreased and cold sensitivity threshold(CST)increased in the model group and the tuina group;after tuina intervention,PMWT in the tuina group was increased,and CST was decreased compared with the model group;after intervention,PMWT in the tuina group was increased,while CST was decreased(P＜0.05).RT-PCR results showed that compared with the sham group,mRNA expression levels of IL-17F,IL-17RC,TRAF6 and Act1 in the spinal dorsal horn of the model group were increased;compared with model group,the mRNA expression levels of above indexes in the tuina group were decreased(P＜0.05).Western boltting results showed that compared with the sham group,the expression levels of IL-17F and TRAF6 protein in the spinal dorsal horn of the model group were increased;compared with the model group,the expression levels of IL-17F and TRAF6 protein in the tuina group decreased(P＜O.05).Immunofluorescence results showed that the mean fluorescence intensity of CD40 in the spinal dorsal horn of model group was enhanced compared with the sham group;compared with the model group,the mean fluorescence intensity of CD40 in the tuina group was decreased(P＜0.05).Conclusion The"three methods and three points"tuina technique can produce immediate analgesia by inhibiting the expression of IL-17F,IL-17RC,Act1,TRAF6 and the activation of M1 microglia in the dorsal horn of the spinal cord after one intervention.

Objective:To analyze the drug-resistant gene loci of Mycoplasma pneumoniae (MP) using metagenomic next-generation sequencing (mNGS). Methods:From November 2022 to October 2023, 697 clinical samples (including sputum, alveolar lavage fluid and blood) of 686 children with Mycoplasma pneumoniae positive detected by mNGS were retrospectively analyzed. Samples were divided into intensive care unit (ICU) group and non-ICU group, Chi-square test was used to compare groups, and Mann-Kendall trend test was used to analyze the change trend of the detection rate of drug resistance gene loci over time. Results:Of the 697 samples, 164 were from the ICU group and 533 were from the non-ICU group. The detection rate of Mycoplasma pneumoniae resistance gene was 44.3% (309/697), and all detected drug-resistant gene loci of MP were A2063G. The detection rate of Mycoplasma pneumoniae in ICU group was 50.0% (82/164), and the detection rates of Mycoplasma pneumoniae resistance gene loci in sputum, alveolus lavage fluid and blood samples were 75.0% (18/24) and 48.4% (62/128), respectively. The detection rate in sputum was higher than alveolus lavage fluid samples ( χ2=5.72, P=0.017). The detection rate of Mycoplasma pneumoniae in non-ICU group was 42.6% (227/533), the detection rate of Mycoplasma pneumoniae resistance gene loci in sputum and alveolar lavage fluid was 40.0% (16/40), 44.3% (201/454), and no detection rate in blood samples (0/12). There was no significant difference in the detection rate of alveolar lavage fluid and sputum ( χ2=0.27, P=0.602). From November 2022 to October 2023, the detection rate of submitted samples showed an increasing trend month by month (overall: Z=3.99, ICU inspection group: Z=2.93, non-ICU group: Z=3.01, all P<0.01). Among the bacteria commonly detected with Mycoplasma pneumoniae, Streptococcus pneumoniae accounted for the highest proportion, the detection rate was 15.5% (108/697), and Epstein-Barr virus accounted for the highest proportion of 17.6% (123/697). Conclusions:From November 2022 to October 2023, the detection rate of Mycoplasma pneumoniae drug resistance gene loci showed an increasing trend. The detection rate of drug resistance gene loci in sputum samples of ICU group was higher than alveolus lavage fluid. No new drug resistance site were detected.

Objective:To assess the effect of mild intraventricular hemorrhage（IVH）on the early motor development of infants at high risk of brain injury,and to guide the intervention according to its characteristics.Methods:A retrospective cohort study was conducted to select neonates discharged from the Neonatal Unit of Xi 'an Children 's Hospital from February 1，2022 to March 31，2023，with one or more high-risk factors of brain injury.The patients were assigned to low-grade IVH group and no IVH group according to ultrasound diagnosis.The research subjects exclucled other brain injury diseases besides mild IVH.Motor development was assessed using test of infant motor performance(TIMP)，reflecting performance in head control，auditory and visual responses，defensive movements，trunk movements，limb movements，and more.Both groups completed TIMP assessment between discharge and 16 weeks of the corrected age(CA).The differences of TIMP scores between two groups were compared . Results:A total of 329 neonates at high risk for brain injury were recruited，including 98 cases with grade Ⅰ-Ⅱ IVH（low-grade IVH group）diagnosed through brain ultrasonography and 231 controls(no IVH group).The Z scores of TIMP in the low-grade IVH group were lower than that in no IVH group（-0.25 ±0.87 vs.0.03 ±0.71， P=0.015）.The risk factors of brain injury were matched for further comparison.At CA2-5 weeks，the scores in low-grade IVH group of TIMP total scores（74.10 ±12.28 vs.84.24 ±7.71），observation items（10.57 ±1.47 vs.11.24 ±1.29），elicitation（63.17 ±12.13 vs.73.00 ±7.36），sitting（9.14 ±2.90 vs.11.65 ±3.26），supine（22.07 ±4.73 vs.24.79 ±3.55），prone position（10.35 ±3.74 vs.12.82 ±3.15）and lateral position（4.00 ±2.85 vs.5.48 ±2.13）were significantly lower than those in no IVH group（ P<0.05）.At CA6-9 weeks，the scores in low-grade IVH group of sitting position（10.44 ±4.01 vs.12.96 ±3.02），supine position（24.04 ±4.60 vs.26.83 ±3.53），lateral position（4.83 ±2.53 vs.6.25 ±2.6）were significantly lower than those in no IVH group（ P<0.05）.At CA12-15 weeks，the low-grade IVH group showed significant differences in TIMP total score（104.00 ±12.98 vs.114.10 ±13.16），elicitation（92.00 ±12.64 vs.102.00 ±13.10），sitting（17.00 ±3.50 vs.19.13 ±3.55）and lateral position（7.35 ±2.14 vs.9.00 ±2.37）compared with those from no IVH group（ P<0.05）. Conclusion:Mild intraventricular hemorrhage affected the early motor development of high-risk infants with brain injury，mainly manifested as a lag in the ability of head control at CA2-5 weeks，and the trend continued until CA12-15 weeks.Early monitoring of motor ability and intervention of head control ability should be carried out in high-risk children with mild intraventricular hemorrhage.

The structure and biological function of CXCL12/CXCR4 are the basis of physiological and pathological function.Combination of HIV-1 envelope protein and CXCR4 will promote the entry of virus into host cells.CXCL12 can reduce the number of CXCR4 through rapid endocytosis and inhibit the replication and transmission of HIV.The interaction of CXCL12/CXCR4 axis with in-flammation and autophagy plays an important role in HIV infection.Previous research has found that gene expression profiles of differ-ent TCM syndromes of acquired immunedeficiency syndrome(AIDS)are different.CXCR4 has different expression in AIDS,lung and spleen qi deficiency syndrome,Qi Yin deficiency syndrome and dampness heat syndrome,which is related to chemokine signaling pathway;the differential gene CXCR4 in peripheral blood of AIDS patients with lung and spleen qi deficiency syndrome is related to au-tophagy process.According to the intervention of Yiaikang Capsule,CXCR4 expression is increased,indicating that Yiaikang Capsule can regulate the expression of autophagy related genes.Research on the role of CXCL12/CXCR4 in TCM syndromes of AIDS is condu-cive to better play the therapeutic advantages of TCM and provide a new direction for gene targeted therapy.

Objective:To systematically evaluate the qualitative research on experience of fear of falling in the elderly, so as to provide theoretical basis for nursing practice.Methods:Qualitative researches on the fear of falling among the elderly were systematically searched in PubMed, Web of Science, Embase, Cochrane Library, China National Knowledge Infrastructure, WanFang Data, VIP, and China Biology Medicine disc. The search period was from database establishment to November 5, 2023. The quality of literature was evaluated using the quality evaluation criteria for qualitative research of the Australia Joanna Briggs Institute Evidence-Based Health Care Center, and the results were integrated using the Meta-synthesis.Results:A total of 13 articles were included, 44 themes were extracted, categorized into 12 categories, and refined into four integrated results, namely perception of fear of falling, influencing factors of fear of falling, coping strategies in attitude and behavior, and multidimensional needs for rehabilitation nursing.Conclusions:Medical and nursing staff should pay attention to and understand the fear of falling among the elderly, evaluate their fear of falling from multiple dimensions and dynamically, provide personalized guidance based on coping behaviors, create support systems to alleviate their fear of falling and meet their multidimensional needs, so as to improve their quality of life.