BACKGROUND:The composite hydrogel scaffold formed by crosslinking of gelatin-methacryloyl(Gel-MA)and treated dentin matrix(TDM)under a certain proportion of ultraviolet light has good porosity,mechanical properties,swelling properties,and biodegradation rate,which provides a new idea and method for clinical pulp regeneration of young permanent teeth. OBJECTIVE:To explore the effect of Gel-MA/TDM composite hydrogel scaffold with 1:2 mass ratio on the proliferation ability and osteogenic/odontoblast differentiation ability of human dental pulp stem cells. METHODS:The passage 3 dental pulp stem cells were inoculated into the Gel-MA/TDM composite hydrogel scaffold with a mass ratio of 1:2.The proliferation ability of human dental pulp stem cells in the composite hydrogel scaffold was detected by CCK-8 assay.Dental pulp stem cells at passage 3 were cultured in Gel-MA/TDM composite hydrogel scaffold with a mass ratio of 1:2 for osteogenic induction.The formation of mineralized nodules was observed by alkaline phosphatase and alizarin red staining.The gene expression levels of odontogenic factors(dentin matrix protein 1,dentin sialophosphoprotein),and osteogenic factors(osteocalcin,Runt-related transcription factor 2)were detected by RT-PCR. RESULTS AND CONCLUSION:(1)The results of CCK-8 assay showed that the proliferation ability of dental pulp stem cells increased significantly in the first 7 days,and slowed down on day 10.(2)The results of alkaline phosphatase staining and alizarin red staining showed that the alkaline phosphatase activity and the formation of mineralized nodules of dental pulp stem cells in the Gel-MA/TDM composite hydrogel group were stronger than those in Gel-MA hydrogel group(P<0.05).(3)RT-PCR results showed that the gene expression levels of dentin matrix protein 1,dentin sialophosphoprotein,osteocalcin,and Runt-related transcription factor 2 in dental pulp stem cells in Gel-MA/TDM composite hydrogel group were significantly higher than those in Gel-MA hydrogel group(P<0.05).The gene expression level at 14 days was significantly higher than that at 7 days(P<0.05).The results conclude that the dental pulp stem cells cultured on Gel-MA/TDM composite hydrogel scaffolds with a mass ratio of 1:2 exhibit a good proliferation ability,which can strengthen the osteogenic and odontogenic differentiation abilities of dental pulp stem cells.

BACKGROUND:Pulp regeneration has been a hot and difficult research topic in recent years,and the construction of composite bio-scaffolding materials provides new ideas and methods for pulp regeneration. OBJECTIVE:To observe the effect of freeze-dried gelatin modified by methacrylic anhydride/treated dentin matrix bioactive scaffolds on proliferation,migration,and osteogenic differentiation of human dental pulp stem cells. METHODS:The mass ratios of gelatin modified by methacrylic anhydride and treated dentin matrix at 2:1,1:1 and 1:2 were obtained by dispersing different masses of treated dentin matrix into gelatin modified by methacrylic anhydride solution.The gelatin modified by methacrylic anhydride/treated dentin matrix bioactive scaffolds were prepared by vacuum freeze-drying.The microstructure,water absorption,and mechanical properties of the scaffolds were measured.Human dental pulp stem cells were cultured with different mass ratios of scaffold extract and DMEM(control group)to detect cell proliferation and migration.Human dental pulp stem cells were cultured with different mass ratios of scaffold extract + osteogenic induction solution and DMEM + osteogenic induction solution(control group),and their osteogenic ability was analyzed by alkaline phosphatase staining. RESULTS AND CONCLUSION:(1)Under scanning electron microscopy,the scaffolds of the three groups all had porous structures.The porosity of the scaffolds increased with the increase of treated dentin matrix quality,and there was significant difference between the two groups(P＜0.05).The water absorption of scaffolds increased with the increase of treated dentin matrix mass,and there was significant difference between groups(P＜0.05).The compressive strength and shear strength of the scaffold increased with the increase of the mass of treated dentin matrix.(2)CCK-8 assay showed that after 3,5,and 7 days of culture,the cell proliferation absorbance values in the 2:1,1:1,and 1:2 scaffold groups were higher than those in the control group(P＜0.05).The cell proliferation absorbance values increased with the increase of treated dentin matrix mass in the scaffold(P＜0.05).The cell scratch test showed that the cell migration rate in the 2:1,1:1,and 1:2 scaffold groups was higher than that in the control group(P＜0.05),and the cell migration rate increased with the increase of treated dentin matrix mass in the scaffold(P＜0.05).(3)Alkaline phosphatase staining showed that the osteogenic differentiation ability of cells in the 2:1,1:1,and 1:2 scaffold groups was stronger than that in the control group,and the osteogenic ability of cells was enhanced with the increase of treated dentin matrix mass in the scaffold.(4)The results showed that the scaffold with a mass ratio of 1:2 between gelatin modified by methacrylic anhydride and treated dentin matrix was the most suitable for the proliferation and differentiation of dental pulp stem cells.

Objective:To investigate the correlation between residual cholesterol (RC) and carotid intima-media thickness (cIMT) in non-diabetic population.Methods:Non-diabetes population received health examination in Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School from January 1, 2018 to December 31, 2021 were retrospectively included. According to the carotid ultrasound results, they were divided into cIMT thickening group (≥1 mm) and non-thickening group (<1 mm). The RC level was calculated according to total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Multivariate logistic regression analysis was used to determine the correlation between RC and cIMT. Results:A total of 1 803 non-diabetes subjects, aged 58.3±15.6 years, including 1 100 males (61.0%), were enrolled. There were 916 patients (50.8%) in the cIMT thickening group and 887 (49.2%) in the non-cIMT thickening group. Compared with the non-cIMT thickening group, cIMT thickening group had significantly higher proportion of male, hypertension, hyperlipidemia, atrial fibrillation, past stroke history, smoking and alcohol consumption, as well as significantly higher age, blood glucose, triglyceride, RC, TC and LDL-C, and significantly lower HDL-C (all P<0.01). Multivariate logistic regression analysis showed that RC was independently associated with cIMT thickening ( Ptrend<0.001). Conclusion:RC is associated with cIMT thickening in non-diabetes population.

Ischemic stroke is a common disease of the nervous system, which is characterized by high incidence, recurrence, disability and mortality rate. The pathological mechanism of ischemic brain injury is complex. Synaptic plasticity injury is considered to be the earliest pathological change after cerebral ischemia, and regulating synaptic plasticity is one of the important mechanisms to promote the recovery of neurological function after stroke. This article reviews the advances in synaptic plasticity after ischemic brain injury, which provides theoretical basis for the development of neuroprotective drugs in the future.

Objective:To investigate the predictors of intracranial hemorrhage in patients with cerebral venous sinus thrombosis (CVST).Methods:Patients with CVST treated in Drum Tower Hospital Affiliated to Medical School of Nanjing University from January 2008 to March 2021 were retrospectively enrolled. The risk factors, clinical manifestations, imaging examination and 90 d follow-up data were collected. The complicated intracranial hemorrhage group and non-intracranial hemorrhage group were compared. Multivariate logistic regression analysis was used to determine the independent predictors of intracranial hemorrhage in patients with CVST. Results:A total of 104 patients with CVST were enrolled, including 42 males and 62 females. Their age was 35.24 ± 10.92 years old (range 22-68 years). Thirty-eight patients (36.84%) were complicated with intracranial hemorrhage, including 34 hemorrhagic cerebral infarction and 4 complicated subarachnoid hemorrhage. Univariate analysis showed that compared with the non-intracerebral hemorrhage group, the intracranial hemorrhage group was more common in puerperal/pregnant patients (60.52% vs. 48.48%; P=0.012), with more acute onset (57.89% vs. 48.48%; P=0.004), focal neurological signs (47.37% vs. 19.70%; P=0.003) and seizure (39.47% vs. 18.18%; P=0.017), and the site of thrombosis was more common in the superior sagittal sinus (57.89% vs. 36.36%; P=0.033). Multivariate logistic regression analysis showed that puerperium/pregnancy (odds ratio 2.857, 95% confidence interval 1.095-7.453; P=0.031) and superior sagittal sinus thrombosis (odds ratio 2.847, 95% confidence interval 1.110-7.302; P=0.027) were the independent predictors of intracranial hemorrhage in patients with CVST. The analysis at 90 d after onset showed that there was no significant difference in the good outcome rate between the intracranial hemorrhage group and the non-intracranial hemorrhage group (86.84% vs. 89.39%; P=0.695). Conclusions:Puerperium/pregnancy and superior sagittalsinus thrombosis are the independent risk factors for intracranial hemorrhage in patients with CVST. However, complicated with intracranial hemorrhage is not associated with 90-day clinical outcomes.

Objective:To compare left and right ventricular Tei indexes and to determine the reference range in newborns of different gestational age (GA) and birth weight (BW).Methods:From February 2019 to June 2021, newborns admitted to the Neonatal Intensive Care Unit of our hospital were enrolled. Tei indexes were measured and calculated during 24 h~7 d after birth and reexamined 1~2 weeks later in some of the newborns. The newborns were assigned into <32 w group, 32~36 w group and ≥ 37 w group according to their GA, < 1 500 g group, 1 500~2 499 g group and ≥2 500 g group according to their BW, and early newborn group (1~7 d) and late newborn group (>7 d) according to their age of evaluation. The data were analyzed using t test, one-way analysis of variance (ANOVA) and correlation analysis with SPSS 20.0 statistical software. Results:A total of 128 cases were included. 42 cases in <32 w group, 43 in 32~36 w group and 43 in ≥37 w group. 42 cases in <1 500 g group, 42 in 1 500 ~ 2 499 g group and 44 in ≥2 500 g group. Tei indexes were reexamined after 7 d of age in 63 preterm infants and in 31 full-term infants. The left and right ventricular Tei indexes of the ≥37 w group were less than the 32~36 w group and the <32 w group in early newborns (left ventricular: 0.382±0.069 vs. 0.431±0.069 and 0.439±0.060, right ventricular: 0.373±0.038 vs. 0.431±0.035 and 0.452±0.064); the right ventricular Tei index of the 32~36 w group was significantly less than the <32 w group ( P<0.05). No significant differences existed in the left ventricular Tei index between the 32 ~ 36 w group and the < 32 w group ( P>0.05). The left and right ventricular Tei indexes of the ≥2 500 g group were significantly less than the 1 500~2 499 g group and the <1 500 g group (left ventricular: 0.385±0.069 vs. 0.434±0.067 and 0.434±0.064, right ventricular: 0.376±0.039 vs. 0.431±0.043 and 0.450±0.061) ( P<0.05).No significant differences existed between the 1 500~2 499 g group and the <1 500 g group ( P>0.05). No significant differences existed in the left and right ventricular Tei indexes between the late newborn group and early newborn group ( P>0.05). For early newborns (1~7 d of age), the reference range of Tei index gradually decreased along with the increase of GA and BW. Conclusions:The left and right ventricular Tei indexes of full-term infants and infants with BW ≥2 500 g are less than preterm and low birth weight infants. The reference range of Tei index in early newborns shows negative correlation with GA and BW.

OBJECTIVE：To inve stigate the effects of polymorphism of CYP3A4，CYP3A5 and other genes on the increasing of tacrolimus blood concentration by Wuzhi capsules in renal transplantation patients. METHODS ：Totally 194 patients in Han nationality underwent renal transplantation and rece ived outpatient follow-up after surgery were selected from the First Affiliated Hospital of Anhui Medical University during Aug. 2015-Nov. 2018，and then divided into single drug group （107 cases）and combination group （87 cases）according to using of Wuzhi capsules or not. Single drug group was given Tacrolimus capsules 0.1-0.15 mg/kg，po，bid+Mycophenolate mofetil capsule s 0.5-0.75 g，bid or Mycophenolate sodium enteric-coated tablets 360-540 mg，bid+Prednisone acetate tablets 10 mg，qd；combination group was additionally given two Wuzhi capsules ，bid，on the basis of single drug group. Two groups were treated at least 7 d. The clinical data （such as patients ’sex and age ）were collected ，and enzyme amplification immunoassay was used to detect steady-state valley concentration of tacrolimus ，and calculate valley concentration/daily dose （C/D）value. PCR was adopted to detect patient genotyping of CYP3A5 gene rs 4646457，rs15524 and rs776746 locus，CYP3A4 gene rs 4646437，rs2242480 and rs 35599367 locus，ABCB1 gene rs 1128503 locus，ABCC2 gene rs3740066 locus，NR1I2 gene rs 3814055 locus，POR gene rs 1057868 locus，PPARA gene rs 4253728 locus，IL-3 gene rs 181781 locus， IL-10 gene rs 1800896 locus，CTLA4 gene rs 4553808 locus，CYP2C19 gene rs 4244285 and rs 4986893 locus，respectively. The correlation of each factor and tacrolimus C/D value was analyzed by Kruskal-Wallis test or Spearman rank test. Multivariate linear regression was conducted. RESULTS ：In 194 renal transplanation patients ，only wild type at rs 3599367 locus of CYP3A4 gene and rs4253728 locus of PPARA gene were detected ，and each genotype distribution of other genes was consistent with the Hardy-Weinberg equilibrium （P＞0.05）. Single factor analysis and multiple linear regression analysis showed that the combination of Wuzhi capsules and rs 776746 polymorphism of CYP3A5 gene were related to tacrolimus C/D value（P＜0.05）. There was statistical significance in tacrolimus C/D values among different genotypes of CYP3A5 gene in single drug group and combnation group（P＜0.05）. C/D value of GG and AG genotype in single drug group were significantly lower than combination group （P＜ 0.05），while there was no statistical significance in tacrolimus C/D value of AA genotype between 2 groups （P＞0.05）. CONCLUSIONS：Combination of Wuzhi capsules or not and polymorphism of CYP3A5 gene rs 776746 locus are associated with blood concentration of tacrolimus in renal transplantation patients in Han nationality. Combined use of Wuzhi capsules can increase blood concentration of tacrolimus in GG and AG genotype ，but have no significant effect on AA genotype.

OBJECTIVE:To explore the clinical related factors of leucopenia induced by azathioprine in the treatment of inflam-matory bowel disease (IBD). METHODS:Clinical information of 114 IBD patients were collected from our hospital during Jan. 2013-Mar. 2015. Steady concentration of AZA metabolite 6-thioguanine(6-TGNs)in red blood cell was determined by HPLC. The correlation of patient’s gender,age,diseases,AZA daily dose and blood concentration of 6-TGNs with leucopenia induced by AZA were investigated. The optimal critical value of leucopenia could be predicted with ROC curves. RESULTS:Among 114 IBD patients,40 patients suffered from leucopenia(35.1%). There was no statistical significance in the proportion of leucopenia among patients with different age,gender,diseases and AZA daily dose(P>0.05). There was statistical significance in the proportion of leucopenia among patients with different concentrations of 6-TGNs(P<0.05). Mean blood concentration of 6-TGNs in leukopenia patients [(407.82±262.88)pmol/(8×108)RBC] was higher than patients with normal leukocyte level [(275.85±118.37)pmol/(8× 108)RBC],with statistical significance(P<0.05). ROC curve predicted that the optimal critical value of leucopenia was blood con-centration of 6-TGNs>291.04 pmol/(8 × 108)RBC. CONCLUSIONS:AZA induced leucopenia may be related to the concentration of 6-TGNs in red blood cell of IBD patients,and high concentration of 6-TGNs is risk factors of leucopenia. Clinicians can provide AZA individual treatment for IBD patient to reduce the occurrence of leucopenia according to routine blood test and the concentra-tion of 6-TGNs.

Objective To develop an HPLC method for the determination of mycophenolic acid(MPA), mycophenolic acid glucuronide(MPAG) in plasma.Methods The samples were precipitated with zinc sulphate-methanol solution before injection.Carbamazepine was selected as internal standard,ZORBAX XDB C18 (4.6 mm ×250 mm,5 μm) column was used and the flow rate was 1 mL/min.The mobile phase consisted of methanol-acetonitrile-potassium dihydrogen phosphate buffer solution(gradient elution) .The column temperature was 30℃ and the detective wave length was 254 nm.And then the MPA,MPAG concentration of 32 patients in 7-14 days after renal transplantion were determined.Results The assay was linear within 0.2-50μg/mL for MPA, 2.5-500 μg/mL for MPAG(r>0.999).Absolute recovery rates of MPA,MPAG were more than 80%, the recoveries were between 90%-110%.The intra-day and inter-day RSDs were both lower than 10%.Totally 32 cases of renal transplantion patiens were with mycophenolate mofetil at the dose of 1-1.5 g/d,and MPA in plasma was within the range of 0.32-6.19μg/mL,MPAG in plasma was within the range of 9.52-149.25μg/mL.Conclusion The method is accurate, convenient and rapid, which could be used in the quantitative determination of plasma concentration of MPA,MPAG in renal transplantion patients.

Objective To prepare a new thrombus model by fluid model method , which is suitable for evaluation of mechanical thrombectomy devices for acute stroke.Methods The fluid model adopted the Chandler loop theory , while the static model adopted conventional method.Mechanical properties of thrombi prepared by fluid model and static model were measured by manual elongation test ( 15 samples in each group) and catheter injection test (15 samples in each group ).Histological structure was compared among the above thrombus models ( 5 samples in each group ) and specimens of five patients with stroke by hematoxylin-eosin staining method.Then, thrombi in fluid model were utilized to establish embolization of vessels in swine model ( two pigs ) for evaluation of radiopacity.Independent-Sample test was performed to compare the maximum tensile length of two methods , and Fisher′s exact test was used to compare the rate of thrombus fragmentation after catheter injection test.Results The maximum tensile length of two models were (4.28 ±0.23) and (3.16 ±0.13) cm, respectively, and the difference was statistically significant (t=16.38, P<0.01);After catheter injection, the fracture rates of thrombus were 13% (2/15) and 60%(9/15), which were also statistically significant (P<0.05).As to histological structure, thrombi prepared by fluid model showed mixed thrombus structure , which similar to thrombi of stroke patients , But thrombi prepared by static model were replete with erythrocytes under the microscope .A total of eight vessels ( lingual arteries and superficial cervical arteries ) in two swines were successfully occlusive ( TIMI 0 or 1 ) , and sufficient radiopacity of each injected thrombus was observed.Conclusions The thrombi prepared by fluid model not only have good mechanical stability and sufficient radiopacity , but their histological structure is similar to thromboemboli retrieved from cerebral arteries of patients with acute stroke.