Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Objective To explore the machine learning model and risk factor analysis for hospital infection caused by carbapenem-resistant enterobacteriaceae(CRE).Methods The clinical data of totally 451 patients infected with extended-spectrum β-lactamases(ESBL)producing Enterobacteriaceae treated in the hospital from 2018 to 2022 were retrospectively collected.The patients were divided into CRE group(115 cases)and sensitive group(336 cases)according to the susceptibility of carbapenem.Four machine learning methods in-cluding Logistic regression analysis,random forest,support vector machine,and neural network were used to build prediction models and receiver operating characteristic curve was used to evaluate.Based on the predic-tion model with the best performance,risk factors for CRE infection were analyzed.Results Random forest model had the best performance,with the area under the curve of 0.952 3.The risk factors for predicting CRE infection by the random forest model included 15 clinical data items,namely fever for more than 3 days,cere-bral injury,drainage fluid sample,trunk surgery,first-level or special-level nursing,ICU treatment,procalcito-nin,anti-anaerobic bacteria,the use of third-generation cephalosporins,age,pre-albumin,creatinine,white blood cell count,and albumin.Conclusion The CRE prediction model developed in this study has good predic-tive value and the risk factors have guiding significance for the early prevention and treatment of CRE infec-tion in clinical practice.

Objective To investigate the effect and mechanism of rhubarb Tangluo pill on liver injury in type 2 diabetic rats.Methods ZDF(fa/fa)rats were given high-fat diet to induce type 2 diabetes model,and were randomly divided into model group,positive control group(0.18 g·kg-1 metformin)and experimental-L,-M,-H groups(0.54,1.08 and 2.16 g·kg-1 rhubarb Tangluo pill),with 8 rats in each group.Eight ZDF(fa/+)rats were selected as control group.The control group and model group were given equal volume of pure water once a day for 12 weeks.An oral glucose tolerance test(OGTT)was performed after administration.Fasting blood glucose level,body mass and liver mass of rats were measured and liver index was calculated.The contents of glutamic-pyruvic transaminase(GPT),glutamic oxalacetic transaminase(GOT),triglyceride(TG)and total cholesterol(TC)in serum were detected.The histomorphologic changes of liver were observed by hematoxylin-eosin(HE)staining and Masson staining.The protein expression of phosphorylated insulin receptor substrate 1(p-IRS1),phosphorylated protein kinase B(p-Akt)and glucose transporter 4(GLUT4)were detected by Western blotting.Results After administration,the fasting blood glucose levels of control group,model group,positive control group and experimental-H group were(4.71±0.45),(29.9±2.97),(15.28±4.52)and(13.84±1.55)mmol·L-1,respectively;the liver index were 2.31±0.46,4.03±0.18,3.37±0.23 and 3.38±0.24;the relative expression level of p-IRS1 protein were 1.00±0.36,4.00±0.11,1.62±0.27 and 1.90±0.17,respectively;the relative expression levels of p-Akt protein were 1.00±0.25,0.21±0.04,0.73±0.15 and 0.54±0.04,respectively;GLUT4 protein relative expression levels were 1.00±0.11,0.40±0.08,0.86±0.04 and 0.70±0.06,respectively.Compared with the model group,the above indexes in the experimental-H group were statistically significant(P＜0.01,P＜0.05).Conclusion Rhubarb Tanglu pill can effectively improve glycolipid metabolism and liver injury in type 2 diabetes mellitus,and its mechanism may be related to the activation of IRS1/Akt signaling pathway.

Objective To investigate the clinical significance of the chorda arteriae umbilicalis in laparoscopic transabdominal preperi-toneal(TAPP)hernia repair.Methods The clinical data of 60 patients with inguinal hernia admitted to Xinrui Hospital in Xinwu District of Wuxi City from June 2019 to June 2022 were analyzed,and the patients were randomly divided into two groups according to whether the chorda arteriae umbilicalis was used as a marker during operation.Both the control group and the observation group were operated according to the routine procedure.The observation group exposed the chorda arteriae umbilicalis,which was used as a reference to precisely free the surgical plane and gap to complete the parietalization of spermatic cord,meanwhile,the angle formed by the intersection of the deferens and umbilical artery cord was used to assist in fixing the patch.The operation time,time of parietalization of spermatic cord,hospital stay,bladder surface bleeding volume,removal rate of hernial sac,the occurrence of postoperative complications and recurrence of patients were compared between the two groups.Results There was no significant difference in the operation time,removal rate of hernial sac,hospital stay,recurrence rate or the incidence of postoperative complications such as chronic pain,uroschesis of patients between the two groups(P>0.05).The time of parietalization of spermatic cord,bladder surface bleeding volume,and incidence of seroma of patients in the observation group were shorter/lower than those in the control group,the differences were statistically significant(P<0.05).Conclusion The chorda arteriae umbilicalis has a constant morphology and relatively fixed anatomical position and alignment.The chorda arteriae umbilicalis can be used as a reference and guiding mark,especially when the anatomical layer is dense and unclear or strayed into the layer in TAPP hernia repair,whihc can guide to operate at the correct layer,standardize the parietalization of spermatic cord,reduce bleeding and vice-damage,and also assist the fixation of the patch and prevent the displacement of the patch.

【Objective】 To explore the effectiveness of using donor plasma reinfusion to flush the leukoreduction system (LRS) chamber during the final reinfusion phase with the Trima Accel automated blood collection system in preventing the reduction of CD4+ and CD8+ T lymphocytes. 【Methods】 A longitudinal and cross-sectional study was designed. CD4+ count<200 cells/μL and CD8+ count<125 cells/μL were considered as the criteria for deficiency. Eighteen first-time platelet donors were followed up. The lymphocyte count was measured at 0, 3-6 and 7-14 times of blood donation in the last 300 days. 170 healthy blood donors who have not donated blood were selected as the control group. According to the cut-off point(October 2021), 88 blood donors who mainly used automatic blood collection system to donate platelet apheresis in the last 365 days(median blood donation times ≥17.5)were divided into three groups(A, B and C)and blood samples were obtained. The time for Groups A, B and C started donating platelet apheresis were as follows: Group A: before October 2019, Group B: from October 2019 to September 2021, Group C: after October 2021. Blood samples were analyzed to obtain blood counts including CD4 + and CD8 + T lymphocytes. Blood samples were analyzed to obtain blood cell counts including CD4+ and CD8+ T lymphocytes. Through a comparative analysis, this study aimed to determine if there are any statistical differences in the detection indices between the follow-up groups with varying frequencies of blood donation, the control group, and groups A, B, and C. This approach was employed to infer the efficacy of donor plasma reinfusion in flushing the leukoreduction system (LRS) chamber for preventing the decline of CD4+ and CD8+ T lymphocytes. 【Results】 Eighteen first-time blood donors who were converted to regular platelet apheresis donors did not show a decrease of CD4 + and CD8 + T lymphocytes in the 5 th and 11 th blood donation (median number of blood donation), and there was no significant difference between the above indexes and those in the 0 th blood donation. Among the previous frequent blood donors, the CD4+ and CD8+ T lymphocyte counts in Group B and Group C are both higher than the standard value, showing no statistical difference from the control group. Among regular blood donors, the CD4+ and CD8+ T lymphocyte counts in groups B and C were higher than the criteria values, and had no statistical difference compared to the control group.The CD4+ T lymphocyte count in Group A was normal, with only one donor in Group A having a CD8+ T lymphocyte count below 125 cells/μL. This donor has donated 281 times of platelet apheresis, and the group he belongs to has started blood donation 2-21 years(median of 5 years) before the adjustment of reinfusion mode. The CD4+ and CD8+ T lymphocyte counts in Group A showed significant differences compared to the control group, with median counts (Group A/Control Group) of 359/521 and 257/372, respectively, P<0.001. In Group A, 0%(0/35) had a CD4+ count below 200 cells/μL, and 2.85%(1/35) of donors had a CD8+ count below 125 cells/μL, which was far lower than the proportion of CD4+ and CD8+ T cell deficiency found in regular apheresis donors by John M. Gansner and Mahboubeh Rahmani. The study showed that the adjustment of the plasma reinfusion mode did not further reduce the T lymphocyte counts in blood donors, but instead further restored the T lymphocyte counts in regular blood donors. This indicated that after the adjustment of plasma reinfusion mode, blood donors might not have lost CD4+ and CD8+ T lymphocytes during blood donation, or only lost a small amount, and can recover even if they donate platelet apheresis frequently. 【Conclusion】 Trima Accel automated blood collection system has a good effect on preventing CD4 + and CD8 + T lymphocytes from being reduced by flushing the LRS chamber with donor plasma.

Proteolysis targeting chimera (PROTAC) is a drug discovery strategy using ubiquitin proteasome system (UPS) to degrade the target protein. Unlike traditional small molecule drugs utilizing occupancy-driven pharmacology as the mode of action (MOA) to regulate protein activity, PROTACs function through forming stable target protein-PROTAC-E3 ubiquitin ligase ternary complex and use ubiquitin proteasome system to degrade the target protein. However, only a few E3 ubiquitin ligases have been used in PROTAC drug design now, and the space of target proteins that PROTAC can target needs to be further expanded. On the other hand, the complicated system of ternary crystal structures is difficult to capture and identify, computational simulation provides modeling of PROTAC-mediated ternary complex formation with effective approaches. In view of this, this review describes the recent progress of bioinformatics on expanding the landscape of E3 ubiquitin ligases and target proteins, and summarizes the methods of computation simulation in modeling PROTAC ternary complex. Finally, the trend of development about PROTAC is prospected.

Pharmaceutical cocrystals is an advanced technology to improve the physicochemical and biological properties of drugs. However, there are few studies on the in vivo metabolism of pharmaceutical cocrystals. In this study, the pharmacokinetics of wogonin cocrystal in normal rats was further studied on the basis of the previous preparation of wogonin-aloperine cocrystal. Firstly, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for simultaneous determination of wogonin and its metabolite wogonoside in rat plasma, and to investigate the methodology. The method was applied to the pharmacokinetic study of wogonin-aloperine cocrystal (Wog-Alop) in rats. The results showed that wogonin and its metabolite wogonoside had a good linear relationship in the range of 1-800 ng·mL^-1, and the precision, accuracy, matrix effect and stability in this range met the requirements of biological analysis. Compared with direct administration of wogonin, the C_max of wogonin and its metabolite in rats increased to 7.44-fold and 9.15-fold, AUC_0-t increased to 1.67-fold and 3.72-fold, and oral bioavailability of wogonin increased to 187.66% after cocrystal administration. Wog-Alop cocrystal can significantly improve the C_max, AUC, and oral bioavailability of wogonin and its metabolite, which provides a new perspective for the clinical application of wogonin. This study was approved by the Experimental Animal Ethics Review Committee of Beijing University of Chinese Medicine (approval number: BUCM-2023032307-1148).

Objective To investigate the clinical efficacy of oblique lumbar interbody fusion(OLIF)combined with poste-rior percutaneous internal fixation in patients with lumbar spinal stenosis with or without redundant nerve roots(RNRs).Meth-ods A retrospective analysis of 92 patients with lumbar spinal stenosis treated by oblique lateral lumbar interbody fusion com-bined with posterior percutaneous internal fixation from June 2019 to June 2022 was performed.There were 32 males and 60 females,aged from 44 to 82 years old with an average of(63.67±9.93)years old.All patients were divided into RNRs positive group and RNRs negative group according to redundancy or not before operation.There were 38 patients in RNRs positive group,including 15 males and 23 females.The age ranged from 45 to 82 years old with an average of(65.45±10.37)years old.The disease duration was 24.00(12.00,72.00)months.There were 54 patients in RNRs negative group,including 17 males and 37 females.The age ranged from 44 to 77 years old with an average of(62.42±9.51)years old.The disease duration was 13.50(9.00,36.00)months.The general data of patients were recorded,including operation time,intraoperative blood loss and complications.The imaging parameters before and after operation were observed,including the number of stenosis segments,intervertebral space height,lumbar lordosis angle and dural sac area.The visual analogue scale(VAS)was used to evaluate the back and lower extremity pain,and the Oswestry disability index(ODI)was used to evaluate the activities of daily living.Results All patients were followed up for 8 to 18 months with an average of(11.04±3.61)months,and no complications were found during the follow-up period.The number of stenosis segments in RNRs positive group(1.71±0.46)was more than that in RNRs negative group(1.17±0.38).In RNRs positive group,intervertebral space height,dural sac area,low back pain VAS,lower extremity pain VAS,ODI score were(1.11±0.19)cm,(0.46±0.17)cm2,(5.39±1.00)scores,(5.05±1.01)points,(55.74±4.05)points,respectively.RNRs negative groups respectively(0.97±0.23)cm,(0.69±0.26)cm2,(4.50±0.77)scores,(4.00±0.58)scores,(47.33±3.43)％.In RNRs positive group,intervertebral space height,dural sac area,low back pain VAS,leg pain VAS,ODI score were(1.60±0.19)cm,(0.74±0.36)cm2,(3.39±0.72)scores,(3.05±1.01)scores,(46.74±4.82)scores,respectively.RNRs negative groups respectively(1.48±0.25)cm,(1.12±0.35)cm2,(3.00±0.82)scores,(3.00±0.82)scores,(37.67±3.58)％.The postoperative intervertebral space height,dural sac area,low back pain VAS score,lower extremity pain VAS and ODI score of the patients in the RNRs positive group and the negative group were signifi-cantly improved compared with those before operation,and the differences were statistically significant(P＜0.05).There were statistically significant differences in the number of stenosed segments,preoperative intervertebral space height,dural sac area,low back pain VAS,lower extremity pain VAS,and ODI between the two groups(P＜0.05).There were significant differences in postoperative intervertebral space height and postoperative ODI between the two groups(P＜0.05),but there was no significant difference in intervertebral space height before and after operation and ODI score before and after operation(P＞0.05).There were significant differences in operation time,intraoperative blood loss,postoperative dural sac area,difference of dural sac area before and after operation,postoperative low back pain VAS,difference of low back pain VAS score before and after oper-ation,difference of lower extremity pain VAS before and after operation between the two groups(P＜0.05).Conclusion OLIF combined with posterior percutaneous internal fixation has a good effect on patients with or without RNRs.Multi-segmental lumbar spinal stenosis and decreased dural sac area may lead to the occurrence of RNRs,and LSS patients with RNRs have more severe symptoms.LSS patients with RNRs have worse surgical outcomes than those without RNRs.

Objective To analyze the relational factors influencing the formation of cauda equina redundant nerve roots(RNRs)of the lumbar spinal stenosis.Methods Clinical data of 116 patients with lumbar spinal stenosis treated from January 2016 to June 2019 were retrospectively analyzed.The patients were divided into redundant nerve roots(RNRs)group and non-RNRs group based on the presence or absence of RNRs on sagittal T2-weighted MRI.In the non-RNRs group,there were 74 patients,including 38 males and 36 females with an average age of(62.00±10.41)years old,the body mass index(BMI)was(23.09±2.22)kg.m-2;the maximum stenosis segment was L2-L3 in 12 cases,L3-L4 in 38,L4-L5 in 20,and L5S1 in 4,respec-tively.In the RNRs group,there were 42 patients,including 18 males and 24 females with an average age of(63.36±8.73)years old,the BMI was(22.63±2.60)kg·m-2;the maximum stenosis segment was L2-L3 in 3 cases,L3-L4 in 9,L4-L5 in 27 and L5S1 in 3,respectively.MRI was performed in the supine position to observe the conshape and morphology of the redundant nerve in the sagittal position.The preoperative low back and leg pain visual analogue scale(VAS),and preoperative Oswestry disability index(ODI)were analyzed,and the rate of spondylolisthesis and ligamentum flavum hypertrophy were compared.Si-multaneously,the inter-vertebral height,intervertebral foramen height,inter-vertebral height+vertebral height,median sagittal diameter at the inter-vertebral space level(DIW-MSD),median sagittal diameter at the pedicel level(DV-MSD),range of mo-tion(ROM)of the stenotic segment were measured and analyzed.Results Among the 116 patients with lumbar spinal stenosis,42 patients developed RNRs,with an incidence of 36.2％.There were no significant differences in gender,age,BMI,preopera-tive VAS for lumbar and leg pain and ODI between two groups(P＞0.05).There were statistically significant differences regard to the duration of symptoms and the rate of spondylolisthesis and ligamentum flavum hypertrophy(P＜0.05);the inter-vertebral height,intervertebral foramen height,inter-vertebral height+vertebral height,DIW-MSD,ROM of the stenotic segment were al-so significantly different between two groups(P＜0.05).However,there was no significant difference in DV-MSD between two groups(P＞0.05).Conclusion The inter-vertebral height,inter-vertebral foramen height,inter-vertebral height+vertebral height,DIW-MSD and ROM of the stenotic segment were the crucial factors related to RNRs in lumbar spinal stenosis.

BackgroundThe guardianship ability of guardians of patients with severe mental disorders plays an important role in supporting the patients' recovery and reintegration into society. It is necessary to develop a scientific tool since there is a lack of tools to quantitatively assess the guardianship ability. ObjectiveTo explore and develop an assessment scale for the guardianship ability of guardians of patients with severe mental disorders， so as to provide references for the construction of scientific and reasonable guardianship ability evaluation tools. MethodsA pool of scale items was constructed through a literature review and interviews， followed by two rounds of expert consultation with 15 specialists. 364 guardians of patients with severe mental disorders in Guangzhou were investigated. The scale items were screened and optimized using item analysis and exploratory factor analysis， and the structural validity of the scale was further verified through confirmatory factor analysis. The content validity of the scale was evaluated by item-level content validity index （I-CVI） the average scale-level content validity index （S-CVI/Ave）. The reliability of the scale was tested by Cronbach's α coefficient and split-half reliability. ResultsThe guardianship ability scale for guardians of patients with severe mental disorders consists of 25 items， including three dimensions of guardianship willingness， guardianship knowledge and behavior and guardianship self-efficacy. The results of the item analysis showed that all items met the corresponding criteria and were retained. Validity test： the I-CVI ranged from 0.800 to 1.000， and the S-CVI/Ave was 0.964. Factor load of each item on the corresponding factors ranged from 0.596 to 0.976， and the model demonstrated good fit： chi-square degree of freedom ratio （χ²/df） was 2.444， Tucker-Lewis index （TLI） was 0.908， comparative goodness of fit index （CFI） was 0.917， standardized root mean square residual （SRMR） was 0.049， and root mean square residual （RMSEA） was 0.089. Reliability test showed that the total scale had a Cronbach's α coefficient of 0.966， and the split half reliability coefficient was 0.915. ConclusionThe guardianship ability scale for patients with severe mental disorders developed in this study has good reliability and validity， and has certain application value for the assessment of guardianship ability for patients with severe mental disorders. ［Funded by Health Science and Technology Project of Guangzhou （number， 20221A011049）］