ObjectiveBased on analytic hierarchy process（AHP）-criteria importance through intercriteria correlation（CRITIC） hybrid weighting method， grey relational analysis and backpropagation artificial neural network（BP-ANN）， to optimize the water extraction process of Qizhi prescription， so as to provide an experimental basis for optimization of the preparation process of this prescription and the establishment of quality standards. MethodsL₉（3⁴） orthogonal test was employed， and the AHP-CRITIC hybrid weighting method was utilized to determine the weight coefficients of the quality fractions of various components， including astragaloside Ⅳ， polygalaxanthone Ⅲ， calycosin-7-O-β-D-glucoside， tenuifolin， and 3，6′-disinapoylsucrose， as well as the dry extract yield. The comprehensive score of each factor level combination in the orthogonal test were calculated as evaluation indicator to select the optimal extraction process parameters. The effects of extraction times， extraction time， and solvent dosage on the aqueous extraction process of the formula were investigated through intuitive analysis， variance analysis， and grey relational analysis. Meanwhile， a BP-ANN model was established to reverse-predict the optimal extraction process parameters of Qizhi prescription， and the optimized process parameters were validated. ResultsThe weight coefficients of the five index components（astragaloside Ⅳ， tenuifolin， calycosin-7-O-β-D-glucoside， polygalaxanthone Ⅲ， and 3，6′-disinapoylsucrose） and dry extract yield were 25.7%， 20.82%， 16.41%， 12.45%， 15.96% and 8.67%， respectively. The optimized extraction process parameters were extracted 3 times with 8， 6， 6 times the amount of water， each time for 1 h. The network prediction results of BP-ANN test samples were consistent with the orthogonal test results， and the mean square error（MSE） of the predicted and measured values of the network was <1%. The water extraction process of Qizhi prescription analyzed and predicted by relevant mathematical models was stable and feasible， which could effectively improve the extraction efficiency of the active ingredients of Astragali Radix and Polygalae Radix， and the average comprehensive score of the validation test was 90.85 with the relative standard deviation（RSD） of 1.55%. ConclusionThis study establishes a water extraction process for compound Qizhi granules， and the optimized extraction process can effectively improve the extraction efficiency of active ingredients， which provides useful references for the optimization of preparation process and the establishment of quality standards for other clinical experience formulas.

BACKGROUND:With the increase of age,the function of bone marrow-derived mesenchymal stem cells is gradually reduced,and delaying the aging of bone marrow-derived mesenchymal stem cells itself has become an important topic.OBJECTIVE:To explore ways to delay the aging of bone marrow-derived mesenchymal stem cells by changing the expression of telomerase Cajal body protein 1(TCAB1)gene.METHODS:Mouse bone marrow-derived mesenchymal stem cells were cultured by cell adhesion method.TCAB1 gene in bone marrow-derived mesenchymal stem cells was overexpressed and interfered by recombinant lentivirus technique.The expression of aging related genes P16,P21,P53,and P27 was detected by qPCR.The relative length of telomeres was detected by qPCR.The expression of aging proteins P16,P21,P53,and P27 was detected by western blot assay.Cell proliferation was detected by CCK-8 assay.Annexin V-PE/7-AAD apoptosis kit was used to detect the degree of cell apoptosis.RESULTS AND CONCLUSION:Bone marrow-derived mesenchymal stem cell lines overexpressing TCAB1 gene had decreased expression of senescence related genes and proteins,increased Telomere relative length,stronger cell proliferation,less apoptosis,and a youthful state.The expression of age-related genes and proteins in bone marrow mesenchymal stem cells interfering with TCAB1 gene increased,and the relative telomere length decreased;cell proliferation ability was weak;cell apoptosis was more,and cells showed senescence.These results indicate that increasing the expression of TCAB1 in an appropriate range can delay the rate of cell senescence.

Objective To investigate the relationship between plasminogen activator inhibitor-1(PAI-1)and mean platelet volume/platelet count ratio(MPV/PLT)with infarct size and cerebral artery stenosis in pa-tients with cerebral infarction.Methods A total of 136 patients with cerebral infarction admitted the hospital from January 2022 to January 2024 were selected as observation group,and another 136 healthy individuals who underwent health check-ups were concurrently selected as control group.136 patients with cerebral in-farction were divided into small infarction group,medium infarction group,and large infarction group based on the infarct size.They were also divided into mild stenosis group,moderate stenosis group,severe stenosis group,and occlusion group based on cerebral artery stenosis degree.PAI-1 level was determined using enzyme-linked immunosorbent assay.Mean platelet volume and platelet count ratio were analyzed by fully automated hematology analyzer,and MPV/PLT was calculated.PAI-1 and MPV/PLT were compared among groups.Then receiver operating characteristic(ROC)curve was plotted to evaluate the value of PAI-1 and MPV/PLT in the diagnosis of cerebral infraction,and the correlation of PAI-1 and MPV/PLT with infarct size and cere-bral artery stenosis degree in patients with cerebral infarction was verified using Spearman correlation analy-sis.Results PAI-1 and MPV/PLT were elevated in observation group compared to control group(P＜0.05).ROC curve analysis yielded that the area under the curve(AUC)of single detection of PAI-1 and MPV/PLT in the diagnosis of cerebral infarction was 0.820(95％CI:0.768-0.873)and 0.754(95％CI:0.695-0.814),while the combined detection of the two reached an AUC of 0.896(95％CI:0.853-0.938).The ser-um PAI-1 level and MPV/PLT in the large infarction group were higher than those in the medium infarction group and small infarction group(P＜0.05),and the levels of the above indicators in the medium infarction group were higher than those in the small infarction group(P＜0.05).The serum PAI-1 level and MPV/PLT in the occlusion group were higher than those in the severe stenosis group,moderate stenosis group and mild stenosis group(P＜0.05),the levels of the above indicators in the severe stenosis group were higher than those in the moderate stenosis group and mild stenosis group(P＜0.05),and the levels of the above indicators in the moderate stenosis group were higher than those in the mild stenosis group(P＜0.05).Spearman corre-lation analysis yielded that PAI-1 and MPV/PLT were positively correlated with infarct size and cerebral ar-tery degree stenosis in patients with cerebral infarction(P＜0.05).Conclusion Patients with cerebral infarc-tion have abnormally elevated PAI-1 and MPV/PLT,and the two indicators are closely correlated with infarct size and cerebral artery stenosis,which have significant potential value in early diagnosis,treatment,and dis-ease assessment of diseases.

BACKGROUND:With the increase of age,the function of bone marrow-derived mesenchymal stem cells is gradually reduced,and delaying the aging of bone marrow-derived mesenchymal stem cells itself has become an important topic.OBJECTIVE:To explore ways to delay the aging of bone marrow-derived mesenchymal stem cells by changing the expression of telomerase Cajal body protein 1(TCAB1)gene.METHODS:Mouse bone marrow-derived mesenchymal stem cells were cultured by cell adhesion method.TCAB1 gene in bone marrow-derived mesenchymal stem cells was overexpressed and interfered by recombinant lentivirus technique.The expression of aging related genes P16,P21,P53,and P27 was detected by qPCR.The relative length of telomeres was detected by qPCR.The expression of aging proteins P16,P21,P53,and P27 was detected by western blot assay.Cell proliferation was detected by CCK-8 assay.Annexin V-PE/7-AAD apoptosis kit was used to detect the degree of cell apoptosis.RESULTS AND CONCLUSION:Bone marrow-derived mesenchymal stem cell lines overexpressing TCAB1 gene had decreased expression of senescence related genes and proteins,increased Telomere relative length,stronger cell proliferation,less apoptosis,and a youthful state.The expression of age-related genes and proteins in bone marrow mesenchymal stem cells interfering with TCAB1 gene increased,and the relative telomere length decreased;cell proliferation ability was weak;cell apoptosis was more,and cells showed senescence.These results indicate that increasing the expression of TCAB1 in an appropriate range can delay the rate of cell senescence.

Objective:To evaluate the accuracy of the domestic "Skywalker" surgical robot in implementing personalized lower limb alignment reconstruction scheme in total knee arthroplasty (TKA) and the short-term clinical outcome of robotic assisted TKA.Methods:From September 2020 to January 2021, the data of patients who received surgical robot assisted TKA in 5 clinical centers in China (Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Southwest Hospital affiliated to Third Military Medical University, The Affiliated Hospital of Qingdao University, The First Affiliated Hospital of Chongqing Medical University and Yantai Yuhuangding Hospital, and Shanghai Ninth People's Hospital affiliated to Shanghai Jiao Tong University School of Medicine was the group leader) were prospectively collected and retrospectively analyzed. There were 24 males and 82 females with an average age of 67.6±7.3 years, (range 45-80 years); Average body mass index 26.42±4.31 kg/m 2, all the operation were performed by "Skywalker" surgical robot system according to preoperative design based on CT. The operation time, intraoperative blood loss, hospitalization days and postoperative complications were recorded, and the imaging indexes including hip-knee-ankle (HKA), lateral distal angle of femur (LDFA) and medial proximal angle of tibia (MPTA) measured before and after the operation, implant model indexes (preoperative planning implant model and postoperative implant model) and short-term clinical efficacy indexes [Western Ontario and McMaster Universities (WOMAC) osteoarthritis index] pain score, stiffness score, joint function score, total score and SF-12 score before and 3 months after the operation) were compared. Results:The average follow-up period was 109.60±9.80 d, (range 95-143 d). The average operation time of 106 patients was 105.30±23.22 min; The average intraoperative blood loss was 141.70±58.33 ml; The average length of hospitalization was 5.82±2.80 d. One patient had ischemic stroke after operation, and one patient had abnormal liver function after operation. According to the judgment of the investigator, all of them were not related to the operation. The actual angle error is the difference between the preoperative planning angle and the postoperative measurement angle. The absolute error of 99.1% (105/106) of the HKA angle was within 3°, 90.8% (69/76) of LDFA, 98.7% (75/76) of the MPTA. In 45 patients in one center where data were available, the actual implant models used in all patients were consistent with the preoperative planning size, and there were only differences in version selection such as Asian condyle. WOMAC pain score, joint function score, total score was improved from 7.34±2.85, 25.10±9.85, 34.75±13.02 to 3.34±2.66, 14.68±9.64, 18.66±13.49 before and after operation, respectively, which were statistically significant ( P<0.001) and SF-12 physiological score and psychological score were improved form 27.24±6.42, 30.68±8.26 to 38.83±5.74, 39.36±7.85 before and after operation, respectively, which were statistically significant ( t=7.33, P<0.001; t=4.53, P=0.043). Conclusion:Domestic surgical robot system "Skywalker" can assist the surgeon to achieve accurate and personalized reconstruction of lower limb alignment and achieve satisfactory short-term clinical outcomes. The long-term clinical outcomes of personalized reconstruction and survival rate of implant still need to be further studied.

Objective:To investigate the influence of joint line alteration on the mid-term clinical effects of primary medial pivot total knee arthroplasty (MP-TKA).Methods:Data of a total 189 patients (233 knees) with 30 males (30 knees) and 159 females (203 knees) with age 66.42±6.12, diagnosed as knee osteoarthritis and underwent primary MP-TKA from January to December 2015, were retrospectively analyzed. All cases were followed up for at least 4 years. The patients were divided into four groups according to the extent of alteration, namely <-4 mm, -4- mm, 0- mm and >4 mm. Range of motion (ROM), Knee Society's Knee Scoring System (KSS), the Western Ontario and McMaster University (WOMAC) osteoarthritis index score were compared preoperatively and at 4 years postoperatively.Results:The postoperative follow-up duration was 52.20± 2.94 months (48-59 months). A total of 133 knee (57%) had joint line raised and 100 cases (43%) joint line distal shifted. The overall height of joint line before surgery was 16.56±3.54 mm (7.33-25.32 mm) and 17.34±2.92 mm (7.25-25.43 mm) postoperatively. The average alteration was 0.78±3.17 mm (-6.00-9.91 mm) with statistically significant difference ( t=-2.596, P=0.010). There was no statistical difference among the four groups in terms of gender, body mass index, pre-operative tibia slope and post condylar offset. The overall preoperative ROM of the all groups was 90.45°±20.44°, which increased from baseline to 108.95°±12.55° at 4 years postoperatively ( t=-22.381, P<0.001). There was statistical difference in term of ROM among each group ( F=7.101, P<0.001). The ROM in group 0- mm and >4 mm was significantly higher than that in group <-4 mm and -4- mm ( P<0.05). The KSS function score and clinic score at 4 years postoperatively were significantly higher that those before surgery, the WOMAC score was statistically lower at 4 years postoperatively ( F=0.723, P<0.001; F=1.138, P<0.001; F=2.502, P<0.001). There was no significant difference among the four groups in terms of KSS score and WOMAC ( P>0.05). One case in >4 mm group had gasket prolapse at 6 month after surgery and 3 cases with immediate anterior knee pain occurred in <-4 mm group. Conclusion:MP-TKA can lead to significant joint line alteration. The overall ROM after surgery is positively correlated with the quantity of joint line alteration. However, there was no correlation between quantity of joint line alteration and the joint function scores. Thus, MP-TKA can achieve good mid-term clinical effects.

Objective: To discuss the effects of sample storage time and temperature on lymphocyte subsets detected by flow cytometry. Methods: Use flow cytometry to detect lymphocyte subsets of a total of 53 blood samples from hospitalized and out-patient patients in Qinghai Provincial People′s Hospital from October 26, 2018 to March 30, 2019. The test was completed within 4 hours after sample collection, which is the control group. The treatment groups are as follows: pretreatment was completed within 4 hours and detected after saving samples at room temperature (group A) for 24 and 36 hours; the tests were performed after keeping samples at room temperature (group B) for 24, 48, 72 hours; completed detection after preserving samples in 4 ℃ condition (group C) for 24, 48, 72 hours. Results: In treatment group A, there was no significant difference in the results of lymphocyte subsets detected after 24 hours of storage compared with the control group(P>0.05); after 36 h of storage, CD3+and CD3+CD8+T lymphocytes percentages were significantly increased [(74.28±11.31)% vs (73.78±11.33)%, (32.15±14.82)% vs (31.00±14.79)%; all P<0.05], while CD19+cells percentage were markedly decreased [(15.60±12.23)% vs (16.11±12.38)%; P<0.05]. In group B, compared with the control group, there was no obvious change in lymphocyte subsets tested after 24 hours(P>0.05); after 48 hours of storage, CD19+cells percentage were observably reduced [(15.60±12.09)% vs (16.11±12.38)%; P<0.05], while other results showed no obvious change (P>0.05); after 72 hours of storage, CD3+and CD3+CD8+T cells percentages elevated significantly [(75.78±11.18)% vs (73.78±11.33)%, (32.57±14.90)% vs (31.00±14.79)%; all P<0.05], and CD19+cells percentage decreased markedly [(14.89±11.92)% vs (16.11±12.38)%; P<0.05]. In group C, there also was no significant change in the results between detecting after 24 hours and the control group(P>0.05); after 48 hours of storage, CD3+CD8+T cells percentage elevated obviously [(32.03±14.95)% vs (31.00±14.79)%; P<0.05] and CD19+cells percentage decreased markedly [(15.32±11.97)% vs (16.11±12.38)%; P<0.05]; after 72 hours of storage, CD3+and CD3+CD8+T cells percentages were significantly increased [(75.63±11.08)% vs (73.78±11.33)%, (32.62±14.98)% vs (31.00±14.79)%; all P<0.05], while CD56+and CD19+cells percentages were reduced observably [(8.21±6.52)% vs (9.02±6.80)%, (14.83±11.79)% vs (16.11±12.38)%; all P<0.05]. Conclusions The blood samples can be kept at room temperature and the testshould be completed within 48 h if the detection of lymphocyte subsets by flow cytometry cannot be performed in time; for the samples that have been pretreated, detection can be completed after saving at room temperature for 24 h, and the above treatments had no significant effect on the inspection results.

Objective To compare the effects of different blood purification methods on blood lipid metabo-lism and malnutrition in maintenance hemodialysis patients. Methods From January 2015 to May 2017,80 patients with maintenance hemodialysis admitted in Wuyi Traditional Chinese Medicine Hospital of Jiangmen were selected. The patients were divided into study group and control group according to the random number table method,with 40 cases in each group. The control group was treated with hemodialysis,the study group was treated with hemoperfusion combined with hemodialysis,all patients were treated for 12 weeks. The levels of total cholesterol (TC),triglyceride (TG),low density lipoprotein cholesterol(LDL - C),hemoglobin(Hb),total protein(TP),albumin(Alb),transferrin (TRF),interleukin 6(IL - 6) and tumor necrosis factor α(TNF - α) were compared before and after treatment. The incidence of complications at the same time were observed. Results After treatment,the levels of TC,TG,LDL - C in the study group were (4. 39 ± 0. 91) mmol/ L,(1. 41 ± 0. 20) mmol/ L,(2. 55 ± 0. 31) mmol/ L,respectively,which were lower than those in the control group [(6. 21 ± 0. 55)mmol/ L,(1. 83 ± 0. 50)mmol/ L,(3. 05 ± 0. 63)mmol/ L] (t = 10. 825,4. 933,4. 504,all P < 0. 05). After treatment,the levels of Hb,TP,Alb,TRF in the study group were (106. 83 ± 22. 05)g/ L,(62. 14 ± 22. 50)g/ L,(38. 30 ± 6. 48) g/ L,(19. 70 ± 2. 20) g/ L,respectively,which were higher than those in the control group [(94. 28 ± 13. 17)g/ L,(52. 38 ± 12. 37)g/ L,(33. 17 ± 6. 80)g/ L,(16. 24 ± 1. 54)g/ L] (t = 3. 090,2. 404,3. 454,8. 146,all P < 0. 05). After treatment,the levels of IL - 6,TNF - α in the study group were (124. 52 ± 107. 23)ng/ L,(72. 13 ± 12. 55)ng/ L,respectively,which were lower than those in the control group [(294. 14 ± 108. 92) ng/ L, (112. 45 ± 21. 29) ng/ L] ( t = 7. 019,10. 318, all P < 0. 05). The incidence rate of complications of the study group was 5. 00％ ,which was lower than 22. 50％ of the control group (χ2 = 5. 165,P < 0. 05). Conclusion Hemoperfusion combined with hemodialysis in maintenance hemodialysis patients can improve their blood lipid metabolism and malnutrition,reduce inflammation and the risk of complications.

BACKGROUND: Studies have shown that umbilical cord mesenchymal stem cells are ideal seed cells for tissueengineering research.OBJECTIVE: To isolate, culture and identify tree shrew umbilical cord mesenchymal stem cells, in order toestablish a standardized tree shrew umbilical cord mesenchymal stem cell lines.METHODS: Caesarean-isolated tree shrew umbilical cord samples were used to isolate and culture umbilical cordmesenchymal stem cells using tissue explant adherent method. Flow cytometry assay was used to detect cellsurface markers. Osteogenic and adipogenic induction media were used to induce umbilical cord mesenchymalstem cells to differentiate into osteoblasts and adipocytes.RESULTS AND CONCLUSION: The cultured umbilical cord mesenchymal stem cells expressed CD90 and CD105 with the positive rate of 99.9% and 99.8% respectively. Hematopoietic stem cell marker CD34 expression ratewas 0.0% and the endothelial cell marker CD31 expression rate was 0.7%, in line with the characteristics of umbilicalcord mesenchymal stem cell surface markers. Calcium nodules by alizarin red staining and lipid droplets by oil red Ostaining were observed in the induced cells. These experimental findings indicated that umbilical cord mesenchymalstem cells from tree shrews capable of osteogenic and adipogenic differentiation were successfully isolated and cultured.

Mercury is one of the common heavy-metal toxins,which can cause damage throughout the body in a variety of ways. Cases of renal toxicity of mercury poisoning are increasing clinically. However,little is known about nephrotoxicity mechanisms,and treatment remains unsatisfactory. The mechanism of mercury toxic nephropathy is reviewed in this paper,including the direct toxic effect on the kidney,the injury to the biomembrane system,generation of Hg-metallothionein,imbalance of intra?cellular calciumion,oxidative damage,induced apoptosis,and immune injury. Besides,the mechanism and limitation of common therapies,potential developments of the field are discussed. This review will facilitate further investigations therapies about both the mechanism and treatment of mercury toxic nephropathy.