Objective:To evaluate the role of spinal cord neuron Src-associated-in-mitosis-68-kDa (SAM68)-transient receptor potential vanilloid-1 channel (TRPV1) signaling pathway in diabetic neuropathic pain (DNP) in mice.Methods:Forty SPF male C57BL/6 mice, aged 8 weeks, weighing 18-22 g, were used in this study. Diabetes mellitus was induced by intraperitoneal streptozotocin 0.12 mg/g, and successful DNP model was defined as decrease in the mechanical paw withdrawal threshold (MWT) of the hind limb≥50% of the baseline value. Twenty-four mice with DNP at 4 weeks after developing the model were divided into 3 groups ( n=8 each) using a simple random sampling: DNP group, SAM68 knocked down group (KD group) and virus control group (VC group). Eight diabetic mice with decrease in MWT <50% were randomly selected as non-DNP group (ND group), and 8 normal mice were randomly selected as control group (NC group). At 4 weeks after developing the diabetes mellitus model, SAM68 gene silencing virus and empty virus were injected into the lumbar enlargement of the spinal cord in KD group and VC group, respectively. MWT was measured before developing the diabetes mellitus model and at 4 and 6 weeks after developing the diabetes mellitus model. The mice were sacrificed after the end of MWT measurement at week 6 after developing the model, spinal cord tissues were collected and the expression of SAM68 and TRPV1 was detected by Western blot, and their localization in the spinal cord was observed by immunofluorescence. Results:Compared with NC and ND groups, the MWT was significantly decreased at 4 and 6 weeks after developing the model, and the expression of SAM68 and TRPV1 in spinal cord tissues was up-regulated in DNP group ( P<0.05). Compared with DNP group, the MWT was significantly increased at 6 weeks after developing the model, the expression of SAM68 and TRPV1 in spinal cord tissues was down-regulated, and no significant change was found in the parameters mentioned above in VC group ( P>0.05). SAM68 and TRPV1 were expressed in neurons in the same region of the spinal cord. Conclusions:Activation of SAM68-TRPV1 signaling pathway in spinal cord neurons is involved in the pathophysiological mechanism of DNP in mice.

Objective:To evaluate the role of lactate-induced mitochondrial division of spinal cord neurons in diabetic neuropathic pain (DNP) in mice.Methods:Thirty-six SPF-grade healthy adult male C57BL/6 mice, aged 2 months, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (CON group), DNP group, and DNP+ sodium oxalate group (OXA group). The diabetic model was established by intraperitoneal injection of streptozotocin 130 mg/kg. After the diabetic model was successfully established, sodium oxalate 750 mg/kg was intraperitoneally injected once a day for 4 consecutive weeks to inhibit lactate production in OXA group, while the equal volume of normal saline was given instead at the same time in C group and DNP group. The mechanical paw withdrawal threshold (MWT) of the left hindpaw was measured before developing the model and at 1, 2, 3 and 4 weeks after developing the model. After completing the last behavioral testing, the spinal cord tissue of the lumbar segment (L 4-6) was taken for determination of the levels of lactate in serum and spinal cord tissues (by the colorimetric method), expression of the mitochondrial membrane potential, reactive oxygen species (ROS) content (using JC-1 or DHE probes), expression of mitochondrial dynamin-related protein 1 (Drp1) and mitochondrial protein mitofusin 2 (Mfn2) (by Western blot), and co-expression of Drp1 and neuronal neuronal marker neuronal nuclear protein (NeuN) (by immunofluorescence double labeling) and for examination of the structure and the number of mitochondria (with a transmission electron microscope). Results:Compared with C group, the MWT was significantly decreased after developing the model, the levels of lactate in serum and spinal cord tissues and ROS content in the spinal cord were increased, the mitochondrial membrane potential was decreased, the Drp1 expression was up-regulated, the Mfn2 expression was down-regulated, the number of mitochondria was increased, the area was reduced ( P<0.05), and the co-expression of Drp 1 and NeuN was increased in DNP group and OXA group. Compared with DNP group, the MWT was significantly increased after developing the model, the levels of lactate in serum and spinal cord tissues and ROS content in the spinal cord were decreased, the mitochondrial membrane potential was increased, the Drp1 expression was down-regulated, the Mfn2 expression was up-regulated, the number of mitochondria was decreased, the area was increased ( P<0.05), and the co-expression of Drp 1 and NeuN was decreased in OXA group. Conclusions:Lactate-induced excessive mitochondrial division of spinal cord neurons can lead to mitochondrial dysfunction, which may be involved in the maintenance mechanism of DNP in mice.

Objective To explore the clinical application effect of scenario-driven unscripted emergency drills.Methods A total of 120 nursing staff who underwent scripted drills from January 2019 to July 2021 were selected as convention group,and 120 nursing staff who underwent scenario-driven unscripted emergency drills at Ganzhou Women and Children's Health Care Hospital from August 2021 to July 2023 were selected as drill group.The two groups were compared in terms of availability of per-sonnel and materials,performance of duties,personnel protection,coordination and organization,division of labor in emergency teams,actual combat effectiveness,partial cooperation and assistance,handling of emergencies,and first aid awareness.Results The drill group demonstrated a higher proportion of personnel who arrived quickly,accurately,and basically on time compared to the convention group.The drill group showed a significantly lower proportion of individual personnel who were not in place and key position personnel who were not in place compared to the convention group(P＜0.05).The drill group performed signifi-cantly better than the convention group(P＜0.05).The drill group did better in on-site material preparation all as well as in per-sonnel protection compared to the convention group.The drill group had a more accurate and efficient overall organization than the conventional group,and the emergency team of the drill group had a more reasonable and efficient division of labor.The drill group demonstrated superior accuracy and efficiency in overall organization compared to the conventional group,with its emergen-cy team exhibiting a more effective and rational division of labor.The drill group achieved their expected outcomes in emergency drills,whereas the convention group achieved only part of their set goals,with most requiring enhancement.The drill group re-ported to superiors in a more timely and efficient manner compared to the convention group,and the group showed better coordina-tion between their departments was good,which could be achieved in a timely manner.The drill group demonstrated a significant-ly better overall response to emergencies and significantly stronger emergency awareness compared to the convention group(P＜0.05).Conclusion Scenario-driven unscripted emergency drills can enhance the emergency response capabilities of nursing staff,improve their decision-making and command skills,optimize organizational coordination,and handling abilities,and strengthen their first aid awareness.

Objective:To evaluate the role of lactate dehydrogenase in diabetic neuropathic pain (DNP) and the relationship with peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) in mice.Methods:SPF-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 18-22 g, were used to establish diabetes mellitus model by intraperitoneal injection of streptozotocin (STZ) 120 mg/kg. Twenty-four mice with diabetes mellitus were divided into 2 groups ( n=12 each) using a random number table method: DNP group and DNP + oxamate group (OXA group). Another 12 SPF-grade healthy male C57BL/6J mice were selected as control group (C group). In OXA group, oxamate 750 mg/kg was intraperitoneally injected once a day for 28 consecutive days. The equal volume of normal saline was given instead in C group and DNP group. The mechanical paw withdrawal threshold (MWT), blood glucose and body weight were measured at 3 days before STZ injection and at 1, 2, 3 and 4 weeks after STZ injection (T 0-4). After the last behavioural test was completed, blood samples were collected from the posterior orbits of anesthetized mice for determination of serum lactate concentrations. The animals were then sacrificed and the tissues from the prefrontal cortex of the brain were taken for determination of lactate content, mitochondrial membrane potential (by the JC-1), content of reactive oxygen species (ROS) (using dihydroethidium probes), and level of histone lactylation and expression of PGC-1α (by Western blot). Results:Compared with C group, the MWT was significantly decreased at T 2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were increased, the mitochondrial membrane potential was decreased, and the expression of PGC-1α was down-regulated in DNP and OXA groups ( P<0.05). Compared with DNP group, no significant change was found in blood glucose and body weight ( P>0.05), the MWT was significantly increased at T 2-4, the serum lactate concentrations, contents of lactate and ROS and level of histone lactylation were decreased, the mitochondrial membrane potential was increased, and the expression of PGC-1α was up-regulated in OXA group ( P<0.05). Conclusions:Lactate dehydrogenase promotes the development of DNP, and the mechanism is related to promotion of increase in histone lactfication and down-regulation of PGC-1α expression in the prefrontal cortex of mice.

Objective This study aimed to investigate the glycosyltransferase gene DsUGT11 involved in the biosynthesis of flavonoids in Desmodium styracifolia,and to analyze the function of its encoding protein by bioinformatic tools,gene cloning,prokaryotic expression,and other technologies.Methods The sequence characteristics and potential biological functions of DsUGT11 were analyzed and predicted by bioinformatics analysis,respectively.Total RNA was extracted from fresh leaves and reverse transcribed into cDNA,from which DsUGT11 gene was successfully amplified and cloned.Heterologous expressed protein was induced and purified,followed by functional characterization using enzymatic reaction in vitro.Results A candidate glycosyltransferase gene,designated as DsUGT11,was identified from the transcriptome data of D.styracifolia.The length of the open reading frame of DsUGT11 is 1426 bp,and the molecular weight of its encoding protein is expected to be 52.14 KDa.By bioinformatic analysis,DsUGT11 was found to harvest a conserved motif of"PSPG"that is unique to the UGT family.Moreover,DsUGT11 was successfully amplified and cloned using the prokaryotic expression vector pMALc5X.Recombinant protein was induced and purified subsequently.Next,the purified protein was used to perform the enzymatic reaction in vitro,the result of which suggested that DsUGT11 was able to catalyze the conversion of 2-OH-naringenin and UDP-glucose into three different compounds,one of which was authenticated as apigenin-7-O-β-D-glucoside(also known as Apigetrin),with two others unknown.Conclusion In this study,the DsUGT11 gene was identified and cloned,whose encoding protein is a flavone-oxyglycosyltransferase catalyzing the conversion of 2-OH-naringenin and UDP-glucose into three different compounds including Apigetrin.

Objective:To evaluate the effect of metformin preconditioning on adenosine monophosphate-activated protein kinase(AMPK)/PTEN-induced putative protein kinase(PINK1) signaling pathway during ischemia-reperfusion (I/R) injury in diabetic rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 6 weeks, weighing 120-160 g, were divided into 3 groups ( n=12 each) by the random number table method: diabetic sham operation group (DS group), diabetic myocardial I/R group (DI/R group) and diabetic myocardial I/R+ metformin preconditioning group(DI/R+ Met group). After 4 weeks of feeding a high-fat and high-glucose diet, the model of type 2 diabetes mellitus was induced by a single intraperitoneal injection of 1% streptozotocin 40 mg/kg. The myocardial I/R injury was induced by blocking the anterior descending branch of the left coronary artery for 30 min followed by 120-min reperfusion in anesthetized animals. In DI/R+ Met group, metformin 200 mg/kg was given by intragastric gavage once a day within 1 week before myocardial ischemia. Blood samples from the femoral vein were collected at 120 min of reperfusion for determination of the serum creatine kinase isoenzymes (CK-MB) and cardiac troponin I (cTnI) concentrations by enzyme-linked immunosorbent assay. Then the rats were sacrificed and myocardial tissues were obtained for examination of the pathological changes(by HE staining) and for determination of the percentage of myocardial infarct size (by the double staining of Ewan blue and TTC) and expression of myocardial autophagy-related protein Beclin-1, PTEN-induced putative kinase 1 (PINK1), phosphorylated 5′-adenosine monophosphate-activating protein kinase (p-AMPK), and ratio of microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ (LC3Ⅱ/Ⅰ) (by Western blot). Results:Compared with DS group, the percentage of myocardial infarct size and serum CK-MB and cTnI concentrations were significantly increased, the expression of Beclin-1, p-AMPK and PINK1 in myocardial tissues was up-regulated, the ratio of LC3II/I was increased( P<0.05), and the pathological changes were aggravated in DI/R group and DI/R+ Met group. Compared with DI/R group, the percentage of myocardial infarct size and serum CK-MB and cTnI concentrations were significantly decreased, the expression of Beclin-1, p-AMPK and PINK1 in myocardial tissues was up-regulated, the ratio of LC3Ⅱ/Ⅰ was increased ( P<0.05), and the pathological changes were significantly reduced in DI/R+ Met group. Conclusions:The mechanism by which metformin preconditioning reduces myocardial I/R injury is related to activation of AMPK/PINK1 signaling pathway and up-regulation of mitochondrial autophagy in diabetic rats.

Humans ; COVID-19 ; Quarantine ; Renal Dialysis ; SARS-CoV-2 ; Hospitals

Objective To investigate the clinical efficacies of free endoscopic nasobiliary drainage (ENBD) in primary duct closure (PDC) following laparoscopic common bile duct exploration (LCBDE) for choledocholithiasis.Methods The retrospective cohort study was conducted.The clinical data of 312 patients with extrahepatic bile duct stones accompanied with or without cholecystolithiasis who were admitted to the 11 medical centers [86 in the Affiliated Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,62 in the Second Affiliated Hospital of Zhejiang University School of Medicine,44 in the West China Hospital of Sichuan University,29 in the First Affiliated Hospital of Xi'an Jiaotong University,27 in the First Hospital Affiliated to Army Medical University (Third Military Medical University),25 in the Hunan Provincial People's Hospital,17 in the Beijing Friendship Hospital of Capital Medical University,10 in the First Affiliated Hospital of Hainan Medical University,5 in the Henan Provincial People's Hospital,4 in the Beijing Tian Tan Hospital of Capital Medical University,3 in the First Affiliated Hospital of Fujian Medical University] from January 2011 to June 2017 were collected.All patients underwent LCBDE+PDC,and 81 and 231 patients with and without ENBD were respectively allocated into the ENBD group and PDC group.Observation indicators:(1) comparisons of operation situations;(2) comparisons of postoperative recovery;(3) comparisons of postoperative complications;(4) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the postoperative complications up to June 2017.Measurement data with normal distribution were represented as x±s.Comparison between groups was analyzed by the t test.Measurement data with skewed distribution were represented M [interquartile range (IQR)],and comparison between groups was analyzed by the nonparametic test.Comparisons of count data were analyzed using the chi-square test and Fisher exact probability.Results (1) Comparisons of operation situations:all the 312 patients underwent successful laparoscopic LCBDE + PDC,without conversion to open surgery,including postoperative death of 1 patient in the PDC group.The common bile duct diameter,cases using interrupted sutures,continuous sutures,absorbable threads and nonabsorbable threads were respectively (1.2±0.4)cm,106,125,195,36 in the PDC group and (1.1±0.5)cm,76,5,79,2 in the ENBD group,with statistically significant differences between groups (t =2.497,x2 =56.706,8.457,P＜0.05).The numbers of stones,stone diameter,cases with common bile duct wall (≤ 3 mm and ＞3 mm),normal and abnormal Oddi sphincter contraction function,volume of intraoperative blood loss and operation time were respectively 2.1±1.7,(1.1-±0.6)cm,148,83,226,5,20 mL (10-45 mL),(116± 49)minutes in the PDC group and 1.9±1.6,(1.0±0.6)cm,49,32,75,6,20 mL (15-30 mL),(113± 23)minutes in the ENBD group,with no statistically significant difference between groups (t =1.021,0.329,x2 =0.329,3.428,Z=1.147,t=0.521,P＞0.05).The further analysis:of 312 patients,cases and time using interrupted sutures and continuous sutures were respectively 182,130 and (133±.49) minutes,(103±34) minutes,with a statistically significant difference between groups (t =-6.605,P＜0.05).The volume of intraoperative blood loss and cases with postoperative complications using interrupted sutures and continuous sutures were respectively 20 mL (15-31 mL),21 and 20 mL (10-45 mL),18,with no statistically significant difference between groups (Z =-0.285,x2 =0.369,P＞ 0.05).Of 312 patients,cases,operation time,volume of intraoperative blood loss and postoperative complications using absorbable threads and non-absorbable threads were respectively 274,(116±44)minutes,20 mL (15-40 mL),33 and 38,(115±35) minutes,18 mL (10-26 mL),6,with no statistically significant difference between groups (Z =0.971,t =0.023,x2 =0.154,P＞ 0.05).(2) Comparisons of postoperative recovery:recovery time of gastrointestinal function,time of abdominal drainage-tube removal,using time of antibiotics and duration of hospital stay were respectively (2.0± 1.5) days,(4.0 ± 2.4) days,(4.0±2.8) days,(5.5±3.0) days in the PDC group and (4.0±1.9) days,(6.9±3.5) days,(10.0± 3.9) days,(11.1±3.7)days in the ENBD group,with statistically significant differences between groups (t =-9.507,-8.258,-15.103,-13.575,P＜0.05).The total expenses of hospital stay in the Affiliated Tongji Hospital,Tongji Medical College of Huazhong University of Science and Technology were respectively (5.1 ±0.6)× 104 yuan in the PDC group and (6.5-±0.5)× 104 yuan in the ENBD group,with a statistically significant difference between groups (t =-9.516,P＜0.05).(3) Comparisons of postoperative complications:incidence of complications in the PDC group was 14.29％ (33/231),including 16 with biliary fistula,11 with biliary tract infection,3 with wound infection,1 with biliary tract bleeding,1 with residual stones of common bile duct and 1 with death;incidence of complications in the ENBD group was 6.17％ (5/81),including 2 with biliary fistula,2 with biliary tract infection and 1 with biliary tract bleeding,showing no statistically significant difference between groups (x2 =3.151,P＞0.05).(4) Follow-up situations:of 312 patients,252 were followed up for 2-67 month,with a median time of 15 months,including 175 in the PDC group and 77 in the ENBD group.During the follow up,there was no occurrence of jaundice,cholangitis and pancreatitis,and stone recurrence and postoperative cholangiostenosis were not detected by abdominal color Doppler ultrasound or CT or magnetic resonanced cholangio-pancreatography.Conclusion On the basis of grasping operative indication strictly,ENBD in PDC following LCBDE for choledocholithiasis is safe and effective.

Objective To investigate the autophagy and apoptosis in acute myelogenous leukemia U937 cell induced by Sirolimus.Methods U937 cells were subcultured, and blank control group(normal) and Sirolimus treated groups(12 h, 24 h,48 h) were established.The Sirolimus treated groups were treated by 2 μmol/L concentration of Sirolimus for 12 h,24 h and 48 h, respectively.The cell morphology of U937 cells treated by Sirolimus was observed after 12 h,24 h and 48 h.The survival rate of cells was detected by cell counting kit-8 method.Cell apoptosis was detected by flow cytometry using Annexin V-FITC/PI double labeled.Real-time PCR was used to detect the level of mRNA expression in autophagy specific protein maker mictotubule-associated protein light chain 3 (LC3)-Ⅱ in different treated times by Sirolimus.Sirolimus LC3 protein expression levels after treatment were detected by Western blot method.Results Under inverted microscope, the cell number of Sirolimus treatment group reduced gradually after 12 h ,24 h and 48 h culture, volume of cells became smaller, cells got ruptured, and the nucleus pycnosis and cellular debris increased.With the extension of time, U937 cells survival rate was falling, and there was statistical differences compared with those of the control group(P =0.031).With Sirolimus treatment, U937 cells after 12 h,24 h and 48 h, U937 cell apoptosis rate increased, and there were statistically significances, compared with those of the control group (P =0.027).With Sirolimus treatment U937 cells after 12 h,24 h and 48 h,LC3-Ⅱ mRNA expression and protein expression were down-regulated compared with those of the control group, and there were statistically significances (P =0.029).Conclusions Sirolimus can induce autophagy and apoptosis in U937 cells.Autophagy protein LC3-Ⅱ in gene and protein expression levels were lowered, and LC3-Ⅱ may play an important role in regulating the leukemia cell autophagy.

BACKGROUND:Cytokines play an important role in the occurrence and development of graft-versus-host disease, but there is a current lack of reports on the association between cytokines and graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation for treatment ofβ-thalassemia major.
OBJECTIVE:To investigate the association between cytokines and graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation forβ-thalassemia major.
METHODS:We observed the dynamic variation of interleukin 6, interleukin 8, interleukin 12, tumor necrosis factor-αand macrophage migration inhibitory factor in 11 children withβ-thalassemia major before onset of graft-versus-host disease, when graft-versus-host disease occurred, at days 4 and 7 after onset of graft-versus-host disease, and when graft-versus-host disease disappeared.
RESULTS AND CONCLUSION:There was a significant difference in serum levels of interleukin-6, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor in different time points, and the highest levels of different cytokines appeared when graft-versus-host disease occurred, fol owed by those at 7 days after
graft-versus-host disease. There was a significant difference in serum levels of interleukin-8 in different time points, and the highest level appeared at 4 days after graft-versus-host disease. The dynamic expression of interleukin-6, interleukin-8, interleukin-12, tumor necrosis factor-α, macrophage migration inhibitory factor can estimate the immune function ofβ-thalassemia major patients who develops graft-versus-host disease after al ogeneic hematopoietic stem cel transplantation, and can be used as the immunobiology indicators for the early diagnosis of graft-versus-host disease.