Objective:To evaluate value of CT perfusion in evaluating the efficacy of different vascular reconstruction methods in the treatment of adult ischemic moyamoya disease.Methods:The clinical data of 80 adult patients with moyamoya disease with ischemic blood type who received revascularization in the Affiliated Hospital of Jining Medical University from February 2019 to February 2023 were retrospectively analyzed, including 43 patients who received superficial temporal arterio-middle cerebral artery anastomosis (direct group) and 37 patients who received brain-dural arterio-artery vascularization (indirect group). The improvement of clinical symptoms, Modified Rankin Scale (mRS) Score, collateral vessels and CTP parameters were compared between the two groups.Results:The improvement of clinical symptoms in the direct group was better than that in the indirect group ( P<0.05), and the significant remission rate was significantly higher than that in the indirect group [86.0%(37/43) vs 59.5%(22/37), χ 2=7.262, P=0.007]. The postoperative mRS score in the direct group was significantly lower than that before surgery [0(0, 1) vs 1(1, 2), Z=5.550, P<0.001]. The score of mRS After surgery in the indirect group was significantly lower than that before surgery [0(0, 1) vs 1(1, 2), Z=4.281, P<0.001], but there was no statistically significant difference between the two groups ( P>0.05). There was no significant difference in digital subtraction angiography (DSA) collateral vessel formation between the two groups ( P>0.05). The CTP parameters of the two groups before and after surgery were consistent between observers and within observers [intraclass correlation coefficient (ICC) range 0.88-0.94]. In the operation area, the relative cerebral blood flow (rCBF) after surgery was significantly increased, and the relative time to peak (rTTP) and relative mean transit time (rMTT) were significantly decreased in both groups, with statistical significance ( P<0.05). In the ipsilateral basal ganglia and frontal lobe, rCBF in the direct group was significantly increased, rTTP and rMTT were significantly decreased, and the differences were statistically significant (all P<0.05), while there were no statistically significant differences in parameters in the indirect group (all P>0.05). The changes of rCBF, rTTP and rMTT in the operative area in the direct group were better than those in the indirect group, and the changes of rCBF, rTTP and rCBF in the basal ganglia and frontal lobe were better than those in the indirect group, with statistical significance (all P<0.05), while there were no statistically significant differences in the remaining parameters (all P>0.05). Conclusions:CT perfusion is of great value in evaluating the difference of clinical symptom improvement in the treatment of adult ischemic moyamoya disease with different vascular reconstruction methods.

Objective To construct an injectable hydrogel(IH)entrapment system of bone marrow mesenchymal stem cells(BM-MSCs)and to explore its effect of promoting articular cartilage repair and underlying mechanism.Methods The primary BM-MSCs of mice were cultured by whole bone marrow adherent method.The differentiation potential was identified by alizarin red(ARS),oil red O(ORO)and Alcian blue(AB)staining,and the expres-sion of CD44,CD90,CD105,CD34 and CD45 on the surface was detected by flow cytometry.BM-MSCs loaded with IH(BM-MSC-IH)was prepared,and the rheological properties of BM-MSC-IH were tested by rheological test.The model of full-thickness injury of knee cartilage in mice was established,and the mice were randomly divided into normal saline group,BM-MSC group and BM-MSC-IH group with 8 in each.The mice were injected with BM-MSC-IH 0.2 mL,cell suspension 0.2 mL and 0.9％NaCl solidion,independently.After 6 and 12 weeks of in-jection,the repair of knee cartilage in mice was observed by safranin-fast green staining,and the expression of typeⅡ collagen in knee joint was examined by immunohistochemical staining microscopy.Results The primary cultured cells showed positive expression of CD44,CD90 and CD105,but negative expression of CD34 and CD45,and had the potential of osteogenic,adipogenic and chondrogenic differentiation.The constructed BM-MSC-IH sys-tem was solid colloid,and the rheological properties were consistent with the basic characteristics of hydrogel.BM-MSC-IH effectively promoted the regeneration of mouse knee cartilage and increase the local expression of typeⅡ collagen.Conclusions Bone marrow-mesenchymal stem cells embedded with injectable hydrogel effectively pro-mote the repair of injured cartilage and the underlying mechanism is potentially promoting the differentiation of chondrocytes and the expression of type Ⅱ collagen.Its effect is better than application of BM-MSCs alone.

Objective:To evaluate value of CT perfusion in evaluating the efficacy of different vascular reconstruction methods in the treatment of adult ischemic moyamoya disease.Methods:The clinical data of 80 adult patients with moyamoya disease with ischemic blood type who received revascularization in the Affiliated Hospital of Jining Medical University from February 2019 to February 2023 were retrospectively analyzed, including 43 patients who received superficial temporal arterio-middle cerebral artery anastomosis (direct group) and 37 patients who received brain-dural arterio-artery vascularization (indirect group). The improvement of clinical symptoms, Modified Rankin Scale (mRS) Score, collateral vessels and CTP parameters were compared between the two groups.Results:The improvement of clinical symptoms in the direct group was better than that in the indirect group ( P<0.05), and the significant remission rate was significantly higher than that in the indirect group [86.0%(37/43) vs 59.5%(22/37), χ 2=7.262, P=0.007]. The postoperative mRS score in the direct group was significantly lower than that before surgery [0(0, 1) vs 1(1, 2), Z=5.550, P<0.001]. The score of mRS After surgery in the indirect group was significantly lower than that before surgery [0(0, 1) vs 1(1, 2), Z=4.281, P<0.001], but there was no statistically significant difference between the two groups ( P>0.05). There was no significant difference in digital subtraction angiography (DSA) collateral vessel formation between the two groups ( P>0.05). The CTP parameters of the two groups before and after surgery were consistent between observers and within observers [intraclass correlation coefficient (ICC) range 0.88-0.94]. In the operation area, the relative cerebral blood flow (rCBF) after surgery was significantly increased, and the relative time to peak (rTTP) and relative mean transit time (rMTT) were significantly decreased in both groups, with statistical significance ( P<0.05). In the ipsilateral basal ganglia and frontal lobe, rCBF in the direct group was significantly increased, rTTP and rMTT were significantly decreased, and the differences were statistically significant (all P<0.05), while there were no statistically significant differences in parameters in the indirect group (all P>0.05). The changes of rCBF, rTTP and rMTT in the operative area in the direct group were better than those in the indirect group, and the changes of rCBF, rTTP and rCBF in the basal ganglia and frontal lobe were better than those in the indirect group, with statistical significance (all P<0.05), while there were no statistically significant differences in the remaining parameters (all P>0.05). Conclusions:CT perfusion is of great value in evaluating the difference of clinical symptom improvement in the treatment of adult ischemic moyamoya disease with different vascular reconstruction methods.

Objective:To assess the clinical value of nomogram based on high resolution magnetic resonance vessel wall imaging (HR-VWI) features in differentiating moyamoya disease (MMD) from atherosclerotic moyamoya syndrome (A-MMS).Methods:Eighty-four patients with digital subtraction angiography (DSA)-confirmed MMD and 73 patients with DSA-confirmed A-MMS were enrolled from Department of Medical Imaging, Affiliated Hospital of Jining Medical University from June 2020 to November 2023. All patients underwent HR-VWI. A retrospective analysis was performed on their imaging data. Univariate analysis was used to compare the differences in imaging characteristics between the two groups. Multivariate Logistic regression analysis was used to screen independent influencing factors for differentiating MMD from A-MMS and a nomogram was constructed accordingly. Receiver operating characteristic (ROC) curve and calibration curve were used to evaluate the validity and calibration of the nomogram.Results:Univariate analysis showed that age, pattern of vessel wall thickening, maximum vessel wall thickness, enhancement degree of vessel wall, vessel external diameter, ipsilateral anterior cerebral artery involvement and dot sign were statistically different between the two groups ( P<0.05). Multivariate Logistic regression analysis showed that age ( OR=6.990, 95% CI: 2.340-20.360, P<0.001), pattern of vessel wall thickening ( OR=0.066, 95% CI: 0.014-0.307, P<0.001), vessel external diameter ( OR=5.224, 95% CI: 1.672-16.324, P=0.005), ipsilateral anterior cerebral artery involvement ( OR=0.160, 95% CI: 0.038-0.679, P=0.013) and dot sign ( OR=0.081, 95% CI: 0.018-0.364, P=0.001) were independent influencing factors for differentiating MMD from A-MMS. ROC curve showed that area under the curve (AUC) of this nomogram was 0.884 (95% CI: 0.821-0.947, P<0.001), and the calibration curve showed a good fit between the predicted probability and actual probability. Conclusion:Nomogram based on HR-VWI features can effectively differentiate MMD from A-MMS.

【Objective】 To investigate the mechanism of Yishen Tonglong Decoction (益肾通癃汤, YSTLD) inhibiting the toll-like receptor 4/p38 mitogen activated protein kinases/nuclear factor kappa-B (TLR4/p38 MAPK/NF-κB) signaling pathway against prostate cancer by up-regulating miR-145-5p. 【Methods】 miRNA microarray technology was used to detect the changes of miRNA expression profile in prostate cancer PC-3 cells treated with YSTLD, and miRNAs with marked differences in miRNA microarray results were screened and validated by real-time polymerase chain reaction (qRT-PCR). Lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells, Cell Counting Kit-8 (CCK8) assay, and scratch assay were adopted to detect the effects of miR-145-5p on prostate cancer PC-3 cell proliferation and migration. qRT-PCR and Western blot were employed to detect the effects of miR-145-5p on TLR4/p38 MAPK/NF-κB signaling pathway and the expression levels of apoptosis-related genes caspase3, tumor necrosis factor-α (TNF-α), Bax, and Bcl-2. qRT-PCR and Western blot were used to detect the effects of serum containing YSTLD on miR-145-5p, TLR4/p38 MAPK/NF-κB signaling pathway, and the expression levels of apoptosis-related genes caspase3, TNF-α, Bax, and Bcl-2. 【Results】 The expression levels of 35 miRNAs in prostate cancer PC-3 cells treated with YSTLD were significantly different from those in the control group, with miR-145-5p being the most significantly different; qRT-PCR validation revealed that the miR-145-5p levels in prostate cancer PC-3 cells treated with YSTLD were significantly higher than those in the DMSO control group (P < 0.05). After lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells, miR-145-5p was found to inhibit the proliferation and migration of prostate cancer PC-3 cells. Overexpression of miR-145-5p up-regulated expression levels of caspase3, TNF-α, and Bax mRNA, and down-regulated expression levels of p38 MAPK, p65 NF-κB, and Bcl-2 mRNA in prostate cancer PC-3 cells (P < 0.05), while up-regulated caspase3 protein expression levels in prostate cancer PC-3 cells and down-regulated expression levels of TLR4, p38 MAPK, and p65 NF-κB protein (P < 0.05). Serum containing YSTLD could up-regulate the expression levels of caspase3, TNF-α, and Bax mRNA, and down-regulate the mRNA expression levels of p38 MAPK, p65 NF-κB, Bcl-2, and TNF receptor-associated factor 1 (TRAF1) in prostate cancer PC-3 cells after intervening prostate cancer PC-3 cells (P < 0.05). Simultaneously, it up-regulated the expression levels of caspase3 protein and down-regulated the protein expression levels of TLR4, p38 MARK, p65 NF-κB, and TRAF1 in prostate cancer PC-3 cells (P < 0.05). 【Conclusion】 YSTLD can promote apoptosis of prostate cancer PC-3 cells by up-regulating the expression level of miR-145-5p and inhibiting TLR4/p38 MAPK/NF-κB signaling pathway, which may be an important mechanism of YSTLD against prostate cancer.

Objective:To explore the feasibility of mismatch of amide proton transfer weighted (APTw) imaging with diffusion weighted imaging (DWI) in evaluating ischemic penumbra (IP) in patients with wake-up stroke.Methods:A prospective study was performed; 96 patients with wake-up stroke and unilateral middle cerebral artery territory infarction admitted to Emergency Stroke Department, Affiliated Hospital of Jining Medical University from September 2020 to January 2023 were chosen. All patients underwent routine MRI, DWI, APTw imaging and 3D arterial spin labeling (3D-ASL) before treatment and 90 d after treatment. IP presence was defined as changes of abnormal signal on T2-fluid-attenuated inversion recovery (FLAIR) 90 d after treatment greater than 20% of high signal range on DWI before treatment, and it was used as the gold standard to compare the efficacy in evaluating whether the patients had IP based on mismatch of 3D-ASLwith DWI and mismatch of APTw imaging with DWI before treatment. The infarct core (IC) region, mismatch region of APTw imaging with DWI, mismatch region of 3D-ASL with APTw imaging were delineated on the fusion images in patients with IP based on mismatch of 3D-ASLwith DWI and mismatch of APTw with DWI, and the differences of APTw values in different regions were compared.Results:According to the 90-d follow-up results, 50 patients had IP and 46 patients did not have IP. Specificity, accuracy and sensitivity evaluating whether the patients had IP based on mismatch of 3D-ASL with DWI were 86.9%, 93.7% and 100.0%, respectively; specificity, accuracy and sensitivity evaluating whether the patients had IP based on mismatch of APTw imaging with DWI were 100.0%, 95.8% and 92.0%, respectively. The APTw max, APTw min and APTw ave values of the IC region were significantly lower than those of mismatch region of APTw with DWI, and the APTw max-min values of mismatch region of APTw imaging with DWI were significantly higher than those of mismatch region of 3D-ASL with APTw imaging ( P<0.05). Conclusion:APTw imaging can reflect the acidosis status of different brain regions in patients with wake-up stroke; specificity and accuracy evaluating whether the patients have IP based on mismatch of APTw imaging with DWI are higher than those based on mismatch of 3D-ASL with DWI.

Objective:To analyze the costs and effectiveness of five common screening modes and genetic screening for thalassemia in China in order to find the optimal way and provide evidence for the implementation of thalassemia prevention and control projects in Hunan Province.Methods:From June 2020 to April 2021, 12 971 couples from 14 cities and autonomous prefectures in Hunan Province were selected as the study population. The diagnosis of thalassemia was based on the results of genetic testing. Results of routine blood test and hemoglobin electrophoresis were collected and analyzed. The efficacy of five screening modes, at the cut-off value of <80 fl or 82 fl for the mean corpuscular volume (MCV), was analyzed by positive predictive value, negative predictive value, Jorden index and cost-effectiveness ratio. Sensitivity analysis was used to assess the feasibility of genetic screening at different costs after fixing the costs of routine blood and hemoglobin electrophoresis. The five thalassemia screening models are as follows: Mode 1: The woman had a blood routine test first. If the result was positive, the spouse required a blood routine test. If both results were positive, a thalassemia gene test should be offered to the couple. Mode 2: Both husband and wife were screened by blood routine and hemoglobin electrophoresis. If one or both of them were positive, both would be tested for thalassemia gene. Mode 3: The couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing. Mode 4: The woman was screened by blood routine and hemoglobin electrophoresis. If any one of them was positive, the woman would be tested for thalassemia gene. If the gene test result was positive, the spouse should receive thalassemia gene. Mode 5: Both spouses conducted a blood routine test. If either was positive, both would conduct hemoglobin electrophoresis test. If both were positive, both spouses should receive thalassemia gene testing. Gene testing mode: The woman would be tested for thalassemia, and her spouse would have thalassemia test too if her result was positive.Results:When using MCV<80 fl as the cut-off for diagnosing thalassemia, the Youden indices of the five prenatal screening modes in Hunan Province were 0.551, 0.639, 0.898, 0.555 and 0.356, while when using MCV<82 fl as the cut-off, the Youden indices were 0.549, 0.629, 0.851, 0.548 and 0.356. When the MCV cut-off value was <80 fl, the missed diagnosis rates of the five screening modes were 44.44%, 0.00, 0.00, 18.52% and 62.96%, and the cost-effectiveness ratios were 21 709, 250 939, 76 870, 138 463 and 92 860 yuan (RMB)/couple, respectively. When the price of genetic testing was lower than 55 yuan (RMB), the cost-effectiveness ratio of genetic screening was lower than that of Mode 3.Conclusions:MCV<80 fl can be considered as the positive criteria in blood routine screening for thalassemia in Hunan Province, and the cost-effectiveness ratio of Mode 3 (the couple received blood routine tests initially. If either was positive, both should receive hemoglobin electrophoresis testing. If either was positive, both parties will conduct thalassemia gene testing) is the best. Genetic screening has certain advantages with the decreasing price.

Objective To compare the preparation efficiency of mouse pox and mouse hepatitis antibodies between two substrains of BALB/c and Big-BALB/c (B-BALB/c) mice, and to provide a theoretical basis and reference for the selection of laboratory animals in the preparation of monoclonal antibodies inducedin vivo through hybridoma.Methods Individuals weighing more than 5% of the weight of normal animals at 4 weeks of age (the criterion for late selection is more than 10%) were selected from a population of conventionally bred BALB/c mice and bred individually, and a subline of B-BALB/c mice was prepared after 10 generations of selection. A total of 40 BALB/c mice and 40 B-BALB/c mice aged 10 to 11 weeks, half male and half female, were selected and inoculated with the mousepox monoclonal antibody hybridoma cell line G23 or the murine hepatitis monoclonal antibody hybridoma cell line Y15 pre-treated with liquid paraffin, respectively. Mice ascites containing monoclonal antibodies were obtained by in vivo induction. The antibody titer was tested by indirect ELISA. The mice were grouped based on the sub-strains, gender and inoculation type of hybridoma to analyze the ascites production, antibody titer and antibody production, and to evaluate the antibody preparation efficiency of the two BALB/c mouse sub-strains.ResultsAfter 10 generations of breeding, the body weight of 10-week-old male and female B-BALB/c mice increased by 22.3% and 12.8%, respectively, compared with BALB/c mice of the same age. Compared with BALB/c mice, B-BALB/c mice had better tolerance and adaptation to secondary ascites collection. Compared with BALB/c mice, the ascites production and antibody titer during the preparation of antibodies in B-BALB/c mice were significantly increased, especially in the hybridoma cell line G23 vaccination group (both P＜0.000 1) . After inoculation with the hybridoma cell lines G23 or Y15, the average antibody production of B-BALB/c mice (14.99×10⁴ U and 33.22×10⁴ U) was higher than that of BALB/c mice (5.33×10⁴ U and 19.31×10⁴ U) (both P＜0.01). After inoculation with hybridoma cell line G23, the average antibody production per unit body weight of B-BALB/c mice (0.55×10⁴ U/g) was higher than that of BALB/c mice (0.23×10⁴ U/g) (P＜0.000 1). And the antibody production per unit body weight of female B-BALB/c or BALB/c mice was higher than that of male B-BALB/c or BALB/c mice (bothP＜0.01).Conclusion B-BALB/c mice can be used as an alternative to BALB/c mice in the in vivo induction of monoclonal antibody preparation, which can achieve the purpose of reducing the number of experimental animals used, lowering the labor cost, and improving the efficiency of antibody preparation.

Objective:To explore the feasibility of CT angiography (CTA) in investigating vascular dilatation of anterior choroidal artery (AChA) and posterior communicating artery (PComA) in patients with Moyamoya syndrome (MMS).Methods:From July 2017 to July 2018, the clinical and imaging data of MMS patients with brain CTA and DSA performed were analyzed retrospectively. According to DSA results, 71 MMS patients were divided into unilateral MMS group (20 cases, 20 hemispheres) and bilateral MMS group (51 cases, 102 hemispheres). There were 20 cases in unilateral MMS group, 10 males and 10 females, with an average age of (45±9) years; 51 cases in bilateral MMS group, 24 males and 27 females, with an average age of (44±12) years. The hemispheres were divided into dilated group and non-dilated group according to the dilatation of AChA or PComA. Kappa analysis was used to evaluate the consistency of two inspection methods to judge the expansion of AChA. The lumen diameters of PComA, P1 and P2 segments of posterior cerebral artery were measured on CTA images, and the ratio of PComA/P1 and PComA/P2 were calculated. The repeatability of CTA measures was evaluated by intra-group correlation coefficient. Independent sample t-test was used to compare CTA measurement results between PComA dilated group and non-dilated group, and ROC curve was drawn to calculate the best threshold for diagnosis of PComA expansion. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of CTA measures were calculated. Results:The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of CTA diagnosis of AChA expansion inunilateral MMS were all 100.00%. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of CTA diagnosis of AChA expansionin bilateral MMS were 90.00%, 93.90%, 93.14%, 78.26% and 97.47%. Compared with DSA, there was no significant difference in the diagnostic performance of AChA expansion between single and bilateral MMS diagnosed by CTA ( P>0.05). The two methods had strong consistency (Kappa value was 1.00 and 0.79 respectively, P<0.01). A total of 46 patients (69 cerebral hemispheres) were included in the evaluation of PComA. PComA/P1 (1.09±0.41) and PComA/P2 (0.86±0.13) in the dilated group were significantly higher than those in the non-dilated group (0.71±0.21 for PComA/P1 and 0.75±0.23 for PComA/P2). The differences were statistically significant ( t=-4.59, -2.50, P<0.05). The best threshold in diagnosing PComA expansion was 0.87 (PComA/P1) and 0.76 (PComA/P2), and the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 84.62%, 83.33%, 84.06%, 86.84%, 80.65% and 79.49%, 60.00%, 71.01%, 72.09% and 69.23%, respectively. Compared with DSA, the Kappa value of CTA measures in diagnosis of PComA expansion was 0.68 (PComA/P1) and 0.40 (PComA/P2), respectively, and the difference was statistically significant ( P<0.05). Conclusions:CTA has a strong consistency with DSA in evaluating the AChA expansion in MMS. When the PComA/P1 ratio on CTA is greater than 0.87, it can be used as the diagnosis criterion for PComA expansion.

Objective:To evaluate the utility of CT perfusion (CTP) for assessment of effect of encephalo-duro-arterio-synangiosis (EDAS) on moyamoya disease (MMD).Methods:Thirty-eight adult MMD patients, who underwent EDAS in our hospital from March 2014 to October 2019, were chosen in our study. All patients received CTP and digital subtraction angiography (DSA) before and after surgery; cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT) and time to peak (TTP) in the operative cerebral cortices and their contralateral mirror areas, as well as brainstems were measured respectively; their relative parameter values (rCBF, rCBV, rMTT and rTTP) were calculated with the brainstem as the reference; perfusion changes of cerebral tissues were observed before and after surgery, and efficacies of these parameters in evaluating the surgical treatment effect were compared. According to the degrees of collateral formation detected by postoperative DSA, the 42 hemispheres of these patients were divided into group of good collateral formation and group of poor collateral formation, and the differences of perfusion changes in the two groups were further discussed.Results:CTP revealed that cerebral perfusion at surgical side after EDAS was significantly improved in all the 42 hemispheres; significantly increased rCBF, and significantly decreased rMTT and rTTP were noted as compared with those before surgery ( P<0.05). Receiver operating characteristic (ROC) curve showed that the area under the curve of rTTP (0.897) was the largest among all parameters, with diagnostic sensitivity of 87.5% and specificity of 80.8%. Twenty-six patients (61.9%) were into the group of good collateral formation and 16 (38.1%) were into the group of poor collateral formation; changed values of rCBF and rMTT in the group of good collateral formation were significantly better than those in the group of poor collateral formation ( P<0.05). Conclusion:CTP can quantitatively evaluate cerebral hemodynamic changes after EDAS in moyamoya disease patients, and postoperative rTTP changes can reflect the degrees of collateral vessel formation.