BACKGROUND:Nerve growth factor is a protein that induces nerve growth and regulates biological behaviors such as proliferation and differentiation of mesenchymal stem cells. OBJECTIVE:To investigate the promoting effect of nerve growth factor on chondrogenic differentiation of bone marrow mesenchymal stem cells. METHODS:Rabbit bone marrow mesenchymal stem cells were isolated and cultured,and nerve growth factor was transfected into bone marrow mesenchymal stem cells by lentiviral transfection.The effects of nerve growth factor on the proliferation,migration,hypertrophic differentiation,and chondrogenic differentiation of bone marrow mesenchymal stem cells were detected by CCK-8 assay,cell scratch assay,alizarin red staining,and western blot assay,using the transfected null-loaded virus as control.To further investigate the promoting effect of nerve growth factor on the chondrogenic differentiation of bone marrow mesenchymal stem cells,interleukin 1β was added in bone marrow mesenchymal stem cells transfected with empty virus and nerve growth factor for 14 days.The expression of proteins related to chondrogenic differentiation and hypertrophic differentiation was detected by western blot assay. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that nerve growth factor had no significant effect on the proliferation of bone marrow mesenchymal stem cells.(2)Compared with the control group,overexpression of nerve growth factor enhanced the migration ability of the cells,and the expression of cartilage-associated proteins type II collagen and SOX9 was up-regulated(P<0.05),while the expression of hypertrophic-associated proteins type X collagen and Runx2 was down-regulated(P<0.05).(3)Compared with the empty virus+interleukin 1β group,the expression of cartilage-associated proteins type II collagen and Sox9 was up-regulated(P<0.05),and the expression of hypertrophy-associated proteins type X collagen and Runx2 was down-regulated after overexpression of nerve growth factor(P<0.05).(4)The results indicated that nerve growth factor could promote the chondrogenic differentiation of bone marrow mesenchymal stem cells.

【Objective】 To investigate the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (M-TESE) in patients with non-obstructive azoospermia (NOA) caused by different causes. 【Methods】 A retrospective analysis was performed on 225 NOA patients during Jan.2020 and Dec.2022. The relation between SRR and patients’ age,body mass index (BMI),testicular volume,endocrine hormones and different etiological classifications were analyzed. 【Results】 According to whether sperm was obtained by surgery,the patients were divided into two groups,including 107 cases in the sperm group and 118 cases in the non-sperm group. There were no significant differences in patients’ age,testicular volume and levels of endocrine hormones between the two groups (P>0.05). According to the different causes,NOA patients with mumps history,cryptorchidism history,AZFc deletion or Klinefelter syndrome (KS) had higher SRR,while idiopathic NOA patients had the lowest SRR (P<0.05). 【Conclusion】 M-TESE is an effective treatment of NOA. There is no correlation between SRR and patients’ age,MBI,testicular volume and levels of endocrine hormones. NOA caused by different etiological classifications may have different SRR.

Objective:To investigate the biomechanical properties of anterior pelvic ring external fixators of two new configurations [iliac crest (IC)+anterior inferior iliac spine (AIIS), anterior superior iliac spine(ASIS)+AIIS] in the treatment of Tile type C1 pelvic fracture.Methods:A 3-dimensional finite element model of Tile type C1 pelvic ring injury (unilateral longitudinal sacral fracture and ipsilateral pubic fracture) was produced. The pelvis was fixed with external fixators of IC, AIIS, combination of IC and AIIS, combination of ASIS and AIIS, and S 1 sacroiliac screw in 5 types of models. In the simulated bipedal standing position and semi-recumbent position, the longitudinal displacement and back rotation angle displacement of the midpoint on the upper surface of S 1 were quantified and compared. Under the simulated left-right compression load state, the lateral displacements of the highest point of the lateral sacral fracture and the highest point of the lateral pubic fracture end were quantified and compared. Under the simulated anterior-posterior shear load state, the backward displacements of the highest point of the lateral sacral fracture end and the highest point of the lateral pubic fracture end were quantified and compared. Results:(1) In the simulated bipedal standing position under the vertical and longitudinal load state, the results of the longitudinal downward displacement of the midpoint on the upper surface of S 1 were consistent with the backward rotation angle displacement, and the order from largest to smallest was IC, AIIS, ASIS+AIIS, IC+AIIS and S 1 sacroiliac screw. The longitudinal downward displacement of IC was significantly larger than that of other models. The longitudinal downward displacement and backward rotation angle displacement of ASIS+AIIS and IC+AIIS were similar, and the latter was smaller. (2) In the simulated semi-recumbent position under the vertical and longitudinal load state, the results of the longitudinal downward displacement and backward rotation angle displacement of the midpoint on the upper surface of S 1 were also consistent, and the order from largest to smallest was IC, AIIS, ASIS+AIIS, IC+AIIS and S 1 sacroiliac screw. (3) Under the simulated left-right compression load state, the results of the lateral displacement of the highest point of the lateral sacral fracture end were consistent with that of the highest point of the lateral pubic fracture end, and the order from largest to smallest was S 1 sacroiliac screw, IC, AIIS, ASIS+AIIS and IC+AIIS. Among them, Among them, The lateral displacement of S 1 sacroiliac screw and IC was larger. The lateral displacement of ASIS+AIIS and IC+AIIS was similar, and the latter was smaller, significantly smaller than that of other models. (4) Under the simulated anterior-posterior shear load state, the results of the backward displacement of the highest point of the lateral sacral fracture end and the highest point of the lateral pubic fracture end were also consistent, and the order from largest to smallest was IC, AIIS, ASIS+AIIS, IC+AIIS and S 1 sacroiliac screw. Among them, the backward displacement of IC and AIIS was larger. The backward displacement of ASIS+AIIS and IC+AIIS was similar, and the latter was smaller. Conclusions:For type C1 pelvic fracture, the biomechanical stabilities of IC+AIIS and ASIS+AIIS are superior to those of IC or AIIS, with ASIS+AIIS being slightly inferior to IC+AIIS. Compared with S 1 sacroiliac screw, IC or AIIS, the lateral stabilities of IC+AIIS and ASIS+AIIS are particularly prominent. The two new external fixator configurations in this study are worthy of clinical application.

Objective:To investigate the prognostic value of baseline peripheral blood inflammatory biomarkers for prognosis in patients with advanced hepatocellular carcinoma (HCC) receiving immunotherapy combined with targeted therapy.Methods:The clinical data of a total of 120 patients with advanced HCC who received immunotherapy combined with targeted therapy at Cancer Center of Renmin Hospital of Wuhan University from December 2019 to March 2022 were analyzed retrospectively. Receiver operating characteristic (ROC) curve was used to calculate the optimal cut-off values of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), systemic immune inflammation index (SII) and prognostic nutritional index (PNI). According to the optimal cut-off values, the study objects were divided into high value group and low value group. The Kaplan-Meier method was used for survival analysis. Cox proportional hazard regression model was applied to analyze the factors associated with prognosis.Results:By the end of follow-up, 74 patients died and 46 survived. The median follow-up time was 23.0 months, the median overall survival (mOS) was 15.6 months, and the median progression-free survival (mPFS) was 13.1 months. ROC curve analysis showed that the optimal cut-off values of NLR, PLR, SII, LMR and PNI were 3.45, 131.87, 626.21, 2.12 and 43.30, respectively. The mPFS (18.3 months vs. 8.7 months) and mOS (26.6 months vs. 10.9 months) of patients in the low-NLR group ( n=75) were longer than those of the high-NLR group ( n=45), and there were statistically significant differences ( χ2=55.64, P<0.001; χ2=64.14, P<0.001). The mPFS (17.9 months vs. 10.9 months) and mOS (24.5 months vs. 13.5 months) of patients in the low-PLR group ( n=55) were longer than those of the high-PLR group ( n=65), and there were statistically significant differences ( χ2=5.27, P=0.023; χ2=11.84, P<0.001). The mPFS (18.0 months vs. 10.7 months) and mOS (25.7 months vs. 12.8 months) of patients in the low-SII group ( n=75) were longer than those of the high-SII group ( n=45), and there were statistically significant differences ( χ2=24.46, P<0.001; χ2=25.42, P<0.001). The mPFS (18.2 months vs. 10.9 months) and mOS (26.6 months vs. 13.2 months) of patients in the high-LMR group ( n=56) were longer than those of the low-LMR group ( n=64), and there were statistically significant differences ( χ2=19.25, P<0.001; χ2=19.92, P<0.001). The mPFS (17.9 months vs. 10.9 months) and mOS (25.4 months vs. 13.4 months) of patients in the high-PNI group ( n=62) were longer than those of the low-PNI group ( n=58), and there were statistically significant differences ( χ2=13.69, P<0.001; χ2=19.07, P<0.001). Univariate analysis showed that Barcelona clinic liver cancer (BCLC) stage ( HR=1.83, 95% CI: 1.17-2.87, P=0.008), Child-Pugh grade ( HR=2.21, 95% CI: 1.47-3.34, P<0.001), modified albumin-bilirubin (mALBI) grade ( HR=1.35, 95% CI: 1.01-1.81, P=0.045), extrahepatic metastases ( HR=2.18, 95% CI: 1.47-3.25, P<0.001), NLR ( HR=1.40, 95% CI: 1.28-1.54, P<0.001), PLR ( HR=1.00, 95% CI: 1.00-1.01, P=0.001), SII ( HR=1.00, 95% CI: 1.00-1.00, P<0.001), LMR ( HR=0.64, 95% CI: 0.51-0.79, P<0.001) and PNI ( HR=0.95, 95% CI: 0.93-0.98, P=0.001) were correlated with PFS; BCLC stage ( HR=2.18, 95% CI: 1.21-3.91, P=0.009), Child-Pugh grade ( HR=2.57, 95% CI: 1.61-4.09, P<0.001), Eastern Cooperative Oncology Group performance status score ( HR=1.59, 95% CI: 1.01-2.51, P=0.044), mALBI grade ( HR=1.60, 95% CI: 1.17-2.17, P=0.003), extrahepatic metastasis ( HR=2.51, 95% CI: 1.59-3.96, P<0.001), NLR ( HR=1.45, 95% CI: 1.32-1.60, P<0.001), PLR ( HR=1.01, 95% CI: 1.01-1.01, P<0.001), SII ( HR=1.01, 95% CI: 1.01-1.01, P<0.001), LMR ( HR=0.57, 95% CI: 0.40-0.72, P<0.001) and PNI ( HR=0.92, 95% CI: 0.89-0.96, P<0.001) were correlated with OS. Multivariate analysis showed that extrahepatic metastasis ( HR=1.78, 95% CI: 1.10-2.87, P=0.018) and NLR ( HR=1.46, 95% CI: 1.24-1.73, P<0.001) were independent influencing factors for PFS; extrahepatic metastasis ( HR=2.09, 95% CI: 1.21-3.61, P=0.009), NLR ( HR=1.56, 95% CI: 1.29-1.88, P<0.001), SII ( HR=1.00, 95% CI: 1.00-1.00, P=0.025), LMR ( HR=0.59, 95% CI: 0.45-0.78, P=0.008) and PNI ( HR=0.93, 95% CI: 0.88-0.99, P=0.013) were independent influencing factors for OS. Conclusion:NLR and extrahepatic metastasis can be regarded as important indicators to predict PFS in patients with advanced HCC receiving immunotherapy combined with targeted therapy, and NLR, SII, LMR, PNI and extrahepatic metastasis can be regarded as important indicators to predict OS in patients with advanced HCC receiving immunotherapy combined with targeted therapy. High NLR, high SII, low LMR, low PNI and extrahepatic metastasis indicate poor prognosis of HCC patients.

OBJECTIVE：To rapidly evaluate the effectiveness，safety and economics of rosuvastatin （RSV）in the treatment of dyslipidemia，so as to provide evidence-based support for clinical drug use. METHODS ：Retrieved from PubMed ，Embase， Cochrane Library ，CNKI，Wanfang database and CBM ，etc.，health technology assessment （HTA）related website and database were searched systematically to select HTA report ，Meta-analysis/systemic evaluation and pharmacoeconomics study about RSV versus placebo or other positive drugs in the treatment of dyslipidemia. According to the inclusion and exclusion criteria ，two researchers independently screened the literatures ，extracted and summarized the data ，then performed qualitative description of results. RESULTS ：Totally 11 Meta-analysis and 11 pharmacoeconomic studies were included ，and no relevant HTA report was retrieved. Results of the study showed that compared with the control group ，RSV could regulate dyslipidemia ，and reduce the levels of LDL-C ，TG，TC，C-reactive protein and sdLDL ；RSV could also reverse atherosclerotic plaque ，reduce all-cause mortality with good safety. In terms of economy ，compared with other statins or placebo ，RSV could prolong quality-adjusted life year，its incremental cost-effectiveness ratio is lower than the desired payment threshold ，which had more economic advantages. CONCLUSIONS：RSV is effective ，safe and economical in the treatment of hyperlipidemia.

Angiogenesis is an essential process in tumor growth, invasion and metastasis. VEGF receptor 2 (VEGFR2) inhibitors targeting tumor angiogenic pathway have been widely used in the clinical cancer treatment. However, most of currently used VEGFR2 kinase inhibitors are multi-target inhibitors which might result in target-associated side effects and therefore limited clinical toleration. Highly selective VEGFR inhibitors are still highly demanded from both basic research and clinical application point of view. Here we report the discovery and characterization of a novel VEGFR2 inhibitor (CHMFL-VEGFR2-002), which exhibited high selectivity among structurally closed kinases including PDGFRs, FGFRs, CSF1R, etc. CHMFL-VEGFR2-002 displayed potent inhibitory activity against VEGFR2 kinase in the biochemical assay (IC = 66 nmol/L) and VEGFR2 autophosphorylation in cells (ECs ∼100 nmol/L) as well as potent anti-proliferation effect against VEGFR2 transformed BaF3 cells (GI = 150 nmol/L). In addition, CHMFL-VEGFR2-002 also displayed good anti-angiogenesis efficacy and exhibited good PK (pharmacokinetics) profile with bioavailability over 49% and anti-angiogenesis efficacy in both zebrafish and mouse models without apparent toxicity. These results suggest that CHMFL-VEGFR2-002 might be a useful research tool for dissecting new functions of VEGFR2 kinase as well as a potential anti-angiogenetic agent for the cancer therapy.

Objective:To prepare a progressive gradient-aperture scaffold composed of silk fibroin(SF)-chitosan(CS)-nano-hydroxyapatite(nHAp)for osteochondral repair.Method:The SF solution, CS solution and nHA suspension were mixed in vitro at equal proportions.The progressive gradient osteochondral(OC)scaffold-1(2%), scaffold-2(3%)and scaffold-3(4%)was respectively prepared by using centrifugation, vacuum freeze-drying, chemical cross-linking and three shaping steps.General conditions, porosity, hot water dissolution rate, water swelling rate, compression water swelling rate, water swelling rate after dissolution, mechanical properties, internal structure observation and pore size were measured.Rat bone marrow mesenchymal stem cells(BMSCs)were cultured and the scaffold extract was prepared.The effect of scaffold extract on the proliferation of BMSCs was detected by the cell counting kit-8(CCK-8)method.BMSCs were co-cultured with the scaffold, and the distribution and morphology of the cells around the scaffold were observed.Results:The structure of scaffold was regular in each group and the porosity was more than 80%.Along with the increase of the material concentration, the water swelling rate of the scaffold was decreased gradually( P<0.05). Compared with before compression, the water swelling rate of scaffold-1 was decreased after compression( P<0.05). There was no significant difference in the hot water dissolution rate among all groups( all P>0.05), and the complete dissolution of the scaffold-1, scaffold-2 and scaffold-3 in vitro required 65.9, 60.9, and 73.9weeks, respectively.The elastic modulus of scaffolds in above three groups were 0.0955, 0.1762 and 0.3468 MPa, respectively.The examination results of scanning electron microscope(SEM)showed that the internal structure of scaffold was honeycomb in each group, the pore shape was regular, which showed an inter-connected pore network.The pore distribution was gradually dense and the pore diameter gradually decreased from the cartilage side to the osteogenic side( P<0.05), and the nHAp content increased gradually.The scaffold extract had no obvious toxicity to the growth and proliferation of BMSCs in each group.After BMSCs were seeded on scaffolds and co-cultured for 5 days, the cells grew well without obvious cell death or morphological abnormalities. Conclusions:In this study, a progressive gradient pore size OC scaffold is successfully prepared with good physical properties and biocompatibility, which is expected to be a new bio-mimetic composite scaffold material for repairing OC defects.

Objective To investigate the expressions of CD28 and CD117 in patients with newly diagnosed multiple myeloma (MM) and their clinical significances. Methods The clinical data of 115 newly diagnosed MM patients in the First Affiliated Hospital of Zhengzhou University from May 2015 to December 2017 were retrospectively analyzed. The expressions of CD28 and CD117 were detected by using multiparameter flow cytometry. The relationship between the expressions of CD28 and CD117 and MM staging and clinical parameters was analyzed. The staging was performed according to the International Staging System (ISS). Results Among these 115 patients, there were 15 patients with CD117 positive and 30 patients with CD28 positive. Erythrocyte sedimentation rate (r ＝ -0.481, P ＝ 0.039), Cˉreactive protein level (r ＝ -0.314, P＝0.015), the proportion of plasma cells detected by bone marrow cytology (r＝-0.027, P＝0.001) were negatively correlated with CD117 positive expressions. CD28 positive expression was positively correlated with lactate dehydrogenase level (r ＝ 0.249, P ＝ 0.033) and ISS stage (r ＝ 0.319, P ＝ 0.017), while it was negatively correlated with hemoglobin level (r ＝ -0.372, P ＝ 0.026). CD28 positive was associated with light chain type, and nonˉsecretory type mostly occurred (P ＝ 0.016). The incidence of osteolytic lesions in CD28 positive group and CD117 positive group was high, but there was no statistical difference between CD28 positive group, CD117 positive group and CD28 negative group, CD117 negative group (P ＝ 0.052, P＝0.479). Conclusions The positive expression of CD117 in the early stage of MM patients is higher than that in the advanced stage, and the expression of CD28 positive in the advanced stage of MM patients is higher than that in the early stage. CD28 and CD117 can be used as indicators of prognosis stratification in the patients with newly diagnosed MM.

Objective To explore the feasibility and methods of preparing silk fibroin(SF)/ chitosan(CS)/nano hydroxyapatite(nHA)composite osteochondral scaffolds with a gradient pore size structure.Methods We prepared an SF solution,a CS solution and an nHA suspension,all with a 2％ concentration,and mixed them with equal proportions.The mixture was used to prepare SF/CS/nHA composite osteochondral scaffolds with a pore size gradient through a centrifugal freeze drying and chemical cross linking method.The porosity,hot water loss rate,water swelling rate and mechanical properties of the scaffold were measured,and the dissolution curve and stress strain curve were drawn.The internal structure and morphology of the scaffold were observed by scanning electron microscopy(SEM)and the sizes of pores in the scaffold were measured.Results The porosity of the scaffold was(91.30± 3.35)％;The five week hot water loss rate was(16.57± 3.18)％;And the water swelling rate was (3218.53 ± 84.37)％.Mechanical test results demonstrated a good compression performance of the scaffold.SEM showed that the internal pores of the scaffolds were honeycomb structured with communicating passages;The density of pores gradually increased with decreasing pore sizes from the top to the bottom(pore sizes at four different levels:(141.11± 11.85)μm,(119.94± 9.05)μm,(93.10 ± 14.98) μm,and (79.95 ± 8.65)μm,respectively,F =22.973,P =0.000).Scaffold cytotoxicity test results indicated no significant difference between A values of the extract group and of the negative control group at any time point(t24 h =0.520,P =0.610;t48 h =0.665,P =0.515;t72 h =0.439,P =0.666),and all RGR values were greater than 100％.Conclusions SF/CS/nHA composite osteochondral scaffolds with a gradient pore size structure can be prepared with a centrifugal-freeze drying and chemical cross-linking method.Scaffolds prepared this way have a three-dimensional structure,a gradient pore size structure,high porosity,strong water absorption,suitable degradation rates and good compressive resistance.Besides,the good cell compatibility and low cytotoxicity of the scaffolds satisfy the requirements for osteochondral tissue engineering materials.

Objective: To investigate the shortage of emergency medicines in emergency department of 76 medical institutions in China, and find out the reasons and the measurements. Methods: The shortage of emergency medicines in 76 class-Ⅰ, class-Ⅱ and class-Ⅲmedical institutions from 22 provinces, autonomous regions and municipalities across the country was investigated by question-naires, and the data obtained from the survey were collected and analyzed by Excel 2007. Results: Totally 76 questionnaires were sent out, and all of them were recycled with the effective recovery of 100% . Pralidoxime chloride injection ranked the first in 59 investiga-ted medicines with the shortage rate of 63. 16% , which was at the top spot in the list of the different medical institutions as well. The main reasons of drug shortage were as follows: less clinical requirements, lost-bid in centralized bidding and price inversion. Conclu-sion: It is suggested that the relevant management departments should establish a reasonable pricing mechanism and carry out policy guarantee for the urgent need of emergency medicines with low dosage and low price.