Biocompatible chitosan/polyethylene glycol/multi-walled carbon nanotube composite scaffolds for neural tissue engineering / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Carbon nanotube (CNT) composite materials are very attractive for use in neural tissue engineering and biosensor coatings. CNT scaffolds are excellent mimics of extracellular matrix due to their hydrophilicity, viscosity, and biocompatibility. CNTs can also impart conductivity to other insulating materials, improve mechanical stability, guide neuronal cell behavior, and trigger axon regeneration. The performance of chitosan (CS)/polyethylene glycol (PEG) composite scaffolds could be optimized by introducing multi-walled CNTs (MWCNTs). CS/PEG/CNT composite scaffolds with CNT content of 1%, 3%, and 5% (1%=0.01 g/mL) were prepared by freeze-drying. Their physical and chemical properties and biocompatibility were evaluated. Scanning electron microscopy (SEM) showed that the composite scaffolds had a highly connected porous structure. Transmission electron microscope (TEM) and Raman spectroscopy proved that the CNTs were well dispersed in the CS/PEG matrix and combined with the CS/PEG nanofiber bundles. MWCNTs enhanced the elastic modulus of the scaffold. The porosity of the scaffolds ranged from 83% to 96%. They reached a stable water swelling state within 24 h, and swelling decreased with increasing MWCNT concentration. The electrical conductivity and cell adhesion rate of the scaffolds increased with increasing MWCNT content. Immunofluorescence showed that rat pheochromocytoma (PC12) cells grown in the scaffolds had characteristics similar to nerve cells. We measured changes in the expression of nerve cell markers by quantitative real-time polymerase chain reaction (qRT-PCR), and found that PC12 cells cultured in the scaffolds expressed growth-associated protein 43 (GAP43), nerve growth factor receptor (NGFR), and class III β‍-tubulin (TUBB3) proteins. Preliminary research showed that the prepared CS/PEG/CNT scaffold has good biocompatibility and can be further applied to neural tissue engineering research.

Added value of 18F-FDG total-body PET/CT dynamic imaging in the diagnosis of liver malignant tumors / 中华核医学与分子影像杂志

Objective:To explore the added value of time-activity curve (TAC) and target-to-background ratio (TBR) obtained by 18F-FDG total-body PET/CT dynamic imaging in the diagnosis of liver malignant tumors. Methods:From December 2019 to October 2021, 109 patients (65 males, 44 females; age (59.3±9.3) years) with hepatocellular carcinoma (HCC; n=27), intrahepatic cholangiocarcinoma (ICC; n=61) and colorectal cancer with liver metastasis (CRLM; n=21) who underwent 60 min 18F-FDG total-body PET/CT dynamic imaging in Zhongshan Hospital, Fudan University were retrospectively enrolled. Dynamic PET/CT images were divided into perfusion-weighted (PW) phase and metabolism-weighted (MW) phase. The arterial phase was defined as the 15 s after the abdominal aorta peak frame at PW. TACs at MW were divided into three types as Graph A, Graph B and Graph C. One-way analysis of variance was used to compare difference of TBR 30/60 among groups. ROC curve analysis was used to evaluate diagnostic effectiveness. Results:With hypervascularity as the diagnostic standard of HCC, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 66.7%(18/27), 75.6%(59/78), 48.6%(18/37) and 86.8%(59/68), respectively. With Graph B as the diagnostic standard of HCC, the sensitivity, specificity, PPV and NPV were 44.4%(12/27), 85.4%(70/82), 50.0%(12/24) and 82.4%(70/85), respectively. The TBR 30/60 of HCC, ICC and CRLM was 0.38±0.19, 0.49±0.18 and 0.64±0.20 respectively ( F=10.89, P<0.001). When the cut-off value of TBR 30/60 was 0.43, the AUC of distinguishing HCC from ICC and CRLM was 0.72, with the sensitivity and specificity of 70.5%(55/78) and 65.2%(15/23). When the cut-off value of TBR 30/60 was 0.64, the AUC of distinguishing ICC from CRLM was 0.71, with the sensitivity and specificity of 61.9%(13/21) and 82.5%(47/57). Conclusion:TAC graph types and TBR 30/60 obtained by total-body PET/CT dynamic imaging display potential value for differentiation between hepatic tumor types.

Preparation and In Vitro Characterization of Gelatin Methacrylate for Corneal Tissue Engineering

BACKGROUND@#Corneal disease is second only to cataract considered as the leading cause of blindness in the world, with high morbidity. Construction of corneal substitutes In Vitro by tissue engineering technology to achieve corneal regeneration has become a research hotspot in recent years. We conducted in-depth research on the biocompatibility, physicochemical and mechanical properties of rat bone marrow mesenchymal stem cells (rBM-MSCs)-seeded gelatin methacrylate (GelMA) as a bioengineered cornea. @*METHODS@#Four kinds of GelMA with different concentrations (7, 10, 15 and 30%) were prepared, and their physicchemical, optical properties, and biocompatibility with rBM-MSCs were characterized. MTT, live/dead staining, cell morphology, immunofluorescence staining and gene expression of keratocyte markers were performed. @*RESULTS@#7%GelMA hydrogel had higher equilibrium water content and porosity, better optical properties and hydrophilicity. In addition, it is more beneficial to the growth and proliferation of rBM-MSCs. However, the 30%GelMA hydrogel had the best mechanical properties, and could be more conducive to promote the differentiation of rBM-MSCs into keratocyte-like cells. @*CONCLUSION@#As a natural biological scaffold, GelMA hydrogel has good biocompatibility. And it has the ability to promote the differentiation of rBM-MSCs into keratocyte-like cells, which laid a theoretical and experimental foundation for further tissue-engineered corneal stromal transplantation, and provided a new idea for the source of seeded cells in corneal tissue engineering.

Investigation and analysis of genetic testing in a SMA cohort based on the national rare diseases registry system of China / 中华检验医学杂志

Objective:To explore application status and development trend of spinal muscular atrophy (SMA) genetic diagnosis technology based on the national rare diseases registry system of China.Method:A total of 200 SMA children registered at the Capital Institute of Pediatrics from July 2016 to December 2018 were included in this retrospective cross-sectional survey. The basic data, clinical subtypes, genotypes, and related genetic testing information of SMA children were obtained by checking SMA registration information, genetic testing reports, and also by telephone follow-up. The patient number and the composition of different genetic diagnosis technologies were analyzed by the stratification of genetic testing at various time. The correlation between the proportion of genetic diagnosis technology and genetic testing time was analyzed with Pearson correlation analysis.Result:There were 3 SMA cases with incomplete data, the remaining 197 SMA cases were included in this study. There were 37 (18.8%), 115 (58.4%) and 45 (22.8%) patients with type Ⅰ, Ⅱ and Ⅲ SMA, respectively. There were 185 cases of SMN1 homozygous deletion (93.9%), and 12 cases with compound heterozygotes (6.1%). Seven SMA-related genetic technologies were used from 2004 to 2017. MLPA accounted for 54.1% (100/185) used approach, followed by PCR-RFLP and first-generation sequencing, which accounted for 22.7% (42/185) and 10.3% (19/185), respectively. Nine, 6, 5 and 4 cases were tested with AS-PCR, qPCR, WES and DHPLC, respectively (2.2%-4.9%). The proportion of MLPA increased gradually since 2010 ( r=0.95, P<0.05), while PCR-RFLP declined gradually since 2004 ( r=-0.99, P<0.05). No correlation was found between technology and testing time for other genetic testing technologies ( P>0.05). The proportion of quantitative genetic technologies (MLPA, qPCR and DHPLC) increased gradually since 2010 ( r=0.94, P<0.05), and qualitative genetic technologies (PCR-RFLP, first-generation sequencing, AS-PCR and WES) decreased gradually since 2004 ( r=-0.94, P<0.05). The duplication detection rates of homozygous deletion and compound heterozygous mutation were 12.4% (23/185) and 41.7% (5/12), respectively (χ 2=5.86, P<0.05). During 2008-2015, the proportion of "the reports of both copy numbers of SMN1 gene and SMN2 gene" increased from 56.8% (21/37) in 2008-2015 to 69.1% (56/81) in 2016-2017. Conclusion:Genetic diagnosis of SMA has gradually developed from qualitative detection technology to quantitative detection technology, such as MLPA and qPCR, in China. In more and more SMA quantitative test reports, quantitative results of SMN2 gene are also provided in addition to quantitative results of SMN1 gene.

Advances in immune checkpoint inhibitor combined with other treatment methods for hepatocellular carcinoma / 临床肝胆病杂志

In recent years, immune checkpoint inhibitor (ICI) in the treatment of unresectable liver cancer has attracted great attention in China and foreign countries and great progress has been achieved, but ICI monotherapy cannot bring benefits to most patients with liver cancer. Therefore, it is a new trend to explore the combination of ICI with other treatment methods. This article summarizes the advances in ICI combination therapy for unresectable liver cancer in China and foreign countries, including ICI combined with molecular targeted therapy, PD-1/PD-L1 inhibitor combined with CTLA-4 inhibitor, and ICI combined with local therapy. The results show that for patients with unresectable liver cancer, ICI combined with other treatment methods has a significantly better effect than ICI monotherapy; however, further studies are needed to explore which treatment method combined with ICI can bring the greatest benefits to patients.

Clinical research of vasospasm and vagus reflex during radial artery puncture treated with auricular point sticking / 中国针灸

<p><b>OBJECTIVE</b>To evaluate the effect and clinical value of auricular point sticking for the diagnosis and treatment of vasospasm and vagus reflex during radial artery puncture, including radial artery spasm (RAS) and coronary artery spasm (CAS).</p><p><b>METHODS</b>A total of 480 patients were randomized into an observation group (224 cases) and a control group (256 cases). Percutaneous coronary intervention and usual care in perioperative period were used in the control group. Auricular point sticking was began to apply 12 h before percutaneous coronary intervention in the observation group at Jiaogan (AH), Shenmen (TF), Pizhixia (AT), Neifenmi (CO), Xin (CO), Shen (CO), Shenshangxian (TG), 1 min a time every point, once every 2 h, 12 h before and after operation. The incidences of vasospasm and vagus reflex during piercing process were compared, and the usage ratios of vasoactive agent were recorded, including glyceryl trinitrate, dopamine and atropine injections.</p><p><b>RESULTS</b>The incidence of angiospasm was 4.9% (11/224) in the observation group, which was lower than 13.3% (34/256) in the control group (<0.01). The incidence of vagal reflex of the observation group was 7.1% (16/224), which was lower than 19.5% (50/256) of the control group (<0.01). The usage ratios of glyceryl trinitrate, atropine and dopamine injections were 3.6% (8/224), 7.1% (16/224), 6.3% (14/224) respectively in the observation group, which were lower than 14.8% (38/256), 15.6% (40/256), 15.2% (39/256) in the control group (all<0.01). .</p><p><b>CONCLUSION</b>Auricular point sticking achieves effect for the diagnosis and treatment of vasospasm and vagus reflex during radial artery puncture.</p>

Distribution of common chromosomal karyotypes in patients with Turner syndrome and correlation between the mean age and height standard deviation scores on diagnosis / 中华实用儿科临床杂志

Objective To analyze the distribution of common chromosomal karyotypes of patients with Turner syndrome (TS), and to explore the correlation between the age and height standard deviation scores (HSDS) on diagnosis.Methods Retrospective investigation was performed for the data of age and HSDS on diagnosis in 273 TS girls(≤ 18.0 years old)diagnosed by chromosomal karyotypes.The main statistical methods were analyzed with t-test and Pearson correlation test by using the SPSS 18.0 statistical software.Results (1) There were 4 kinds of common chromosomal karyotypes in the TS :45, X (87/273 cases,31.9％),46, X, i (Xq) (43/273 cases, 15.7％) ,45, X/46, X, i (Xq) (36/273 cases, 13.2％) and 45, X/46, XX (23/273 cases, 8.4％), respectively, the adolescent TS all had delayed puberty.For the cases with 45, X karyotypes ,3 cases presented mental retardation and 2 cases with organs deformity.(2)The patients with 45 ,X/46,X,i(Xq) karyotypes or with 46,X,i(Xq) karyotypes had the maximum(12.56 age) or the minimum(9.70 age) mean age on diagnosis, respectively, there was a significant difference between 2 groups (t =3.019, P =0.004).The maximum deviation from normal height was found in the patients with karyotypes of 46, X,i (Xq) (HSDS =-4.04), and the minimum deviation was in the patients with karyotypes of 45,X/46, XX (HSDS =-3.16), and there was a significant difference between 2 groups (t =-2.95, P =0.004).(3) More than 75.7％ of TS patients was diagnosed when their heights deviated above 3 SD,and their mean age on diagnosis was 12.10 age,which was 3 years later than those patients within 2 SD.(4) There was a significant negative correlation between the age and HSDS on diagnosis in the groups of common chromosomal karyotypes[45,X、46,X,i(Xq) and 45,X/46,XX] (r =-0.551,-0.560,-0.622,all P ＜ 0.01), except for the group with the 45, X/46, X, i (Xq).Conclusions (1) In this study, the consti-tuent ratios of these 4 common chromosomal karyotypes were different from those in Europe and America's.(2)Patients with 45 ,X may have more severe symptoms than others.(3)The mean age on diagnosis was at least 3.0 years earlier when considered HSDS below-2.00 as an indicator for chromosomal karyotype screening,which would facilitate earlier diagnosis.

Screening for genetic mutations in hyperphenylalaninemia using Ion Torrent PGM sequencing / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a hyperphenylalaninemia related genes screening method using Ion Torrent Personal Genome Machine (PGM) for early detection and differential diagnosis of hyperphenylalaninemia (HPA).</p><p><b>METHODS</b>Three children with known HPA mutations and a healthy control were used for setting up the method. Ten children with HPA with known mutations were recruited for validating the method. Ion Ampliseq PCR was used to amplify the 5' and 3' untranslated region, coding sequence, and flanking introns of PAH, GCH1, PTS, QDPR, and PCBD1 genes. After the enrichment with the Ion OneTouch system, the products were sequenced by PGM. Data from the PGM were processed with Torrent Suite v2.2 software package. All variations were confirmed by Sanger sequencing.</p><p><b>RESULTS</b>For the 4 samples, the PGM output was 94.22 Mb, with approximately 99.5% of reads mapping to the target regions. Among these samples, we detected 74 variations (28 positions) including 6 known mutations. Compared with database and results of Sanger sequencing, 55 (18 positions) polymorphisms and 13 (4 positions) false positive calls were confirmed. For the 10 samples, all the known mutations were successfully identified.</p><p><b>CONCLUSION</b>Ion Torrent PGM sequencing is suitable for screening genetic mutation underlying HPA from the perspective of metabolic pathways, which can meet the clinical demand for individualized diagnosis and treatment.</p>

Effect of intrathecal nerve growth factor on lidocaine-induced neurotoxicity to spinal cord in rats / 中华麻醉学杂志

Objective To investigate the effect of intrathecal nerve growth factor (NGF) on lidocaineinduced neurotoxicity to the spinal cord in rats.Methods Thirty healthy male Sprague-Dawley rats,aged 8-10 weeks,weighing 250-300 g,were randomly divided into 5 groups (n =6 each) using a random number table:control group (group C),sham operation group (group S),lidocaine-induced neurotoxicity group (group L),normal saline group (group NS) and NGF group (group NGF).A catheter was inserted at L4.5 interspace into the epidural space in S,L,NS,and NGF groups.One day after surgery,20％ lidocaine 20 μl was injected intrathecally.At 24 h after lidocaine injection,normal saline 20 μl and NGF 10 μg (20 μd) were injected intrathecally in NS and NGF groups,respectively,once a day for 7 consecutive days.The tail flick latency (TFL) to a thermal nociceptive stimulus was measured on 1,3,5 and 7 days after lidocaine injection.The animals were sacrificed after the behavioral test was completed at 7 days after lidocaine injection,and the lumbar segments of the spinal cord were removed to detect the neuronal apoptosis (using flow cytometry) and caspase-3 mRNA expression (by RT-PCR).Results Compared with group C,the TFL was significantly prolonged at 1-7 days after lidocaine injection in L and NS groups and at 1-5 days after lidocaine injection in group NGF,the apoptosis rate was increased in L,NS and NGF groups,and caspase-3 mRNA expression was up-regulated in L and NS groups,and no significant change was found in the parameters mentioned above in group S.Compared with group L,the TFL was significantly shortened at 5 and 7 days after lidocaine injection,the apoptosis rate was decreased,and caspase3 mRNA expression was down-regulated in NGF group,and no significant change was found in the parameters mentioned above in group NS.Conclusion Intrathecal NGF can reduce lidocaine-induced neurotoxicity to the spinal cord in rats and inhibition of caspase-3 mRNA expression is involved in the mechanism.

Identification of two survival motor neuron gene 1 gene mutations and evaluation of their effects on full-length survival motor neuron gene 1 transcripts / 中华神经科杂志

Objective To perform mutation analysis of survival motor neuron gene 1 (SMN1 in two spinal muscular atrophy (SMA) patients and their parents to evaluate the effects of the two SMN1 gene mutations on the transcript levels of the gene and preliminarily predict their effects on the structure and function of SMN protein.Methods Mutation analysis of SMN1 gene was carried out by multiplex ligationdependent probe amplification,reverse transcript-polymerase chain reaction (RT-PCR) and cloning sequencing.Transmission of the mutations was confirmed by the mutation analysis in patients' parents.The full-length SMN1 (SMN1-fl) transcript levels of the patients carrying these subtle mutations were detected using quantitative RT-PCR.Results The two patients were diagnosed as SMA Ⅱ and SMA Ⅲ.They carried p.Val19GlyfsX21 and p.Ala2Gly SMN1 mutations in SMN1 gene,respectively.Both of the two mutations were originated from their fathers.Compared with the healthy individuals (23.5 ± 4.9),the two patients had a significant reduction in the level of SMN1-fl transcripts (t =3.322,P =0.011 (p.Ala2Gly) ;t =6.964,P =0.000 (p.Val19GlyfsX21)).However,compared with the healthy carriers (14.1 ±4.5),the patient with p.Ala2Gly mutation had no significant reduction in the level of SMN1-fl transcripts (13.9 ±3.6,t =0.058,P =0.955) ; however,the patient with p.Val19GlyfsX21 mutation had a significant reduction (4.9± 2.4,t =3.725,P =0.004).Conclusions Two SMN1 gene mutations are identified in our study.The mutation p.Val19GlyfsX21 is a novel mutation and p.Ala2Gly is firstly reported in Chinese SMA patients.p.Val19GlyfsX21 may possibly lead to decreased SMN1-fl mRNA by nonsense-mediated messenger RNA decay,however,p.Ala2Gly has no obvious effects on the amount of the SMN1-fl transcripts,indicating that its deleterious effect may be occurring at SMN protein level or the function of SMN protein.

Progress in molecular Biology of MELAS / 基础医学与临床

MELAS syndrome(mitochondrial myopathy encephalophathy with lactic acidosis and stroke-like episodes),as one of the most common diseases in mitochondrial encephalomyopathies,is characterized by highly variable manifestations.So,more and more people come to realize the importance of molecular basis of MELAS.This review took the commonest mtDNA point mutation(A3243G) for example to overview its molecular biological mechanism,test strategy and recent progress of study on MELAS syndrome.

Study on Preparation and Quality Standard of Ethacridine Lactate Solution / 中国药房

OBJECTIVE: To prepare ethacridine lactate solution and to evaluate its quality.METHODS: The solution was prepared with ethacridine lactate as principal components and sterilized by steam sterilization.HPLC method was used for the determination of ethacridine.Calesil ODS column was used with mobile phase consisted of methanol-acetonitrile-0.05% sodium laurysulfonate (pH=3.0,20 ∶ 20 ∶ 60) at detection wavelength of 270 nm.The column temperature was set at 30 ℃ and injection volume was 10 ?L.The effect of sterilization on the content of preparation was determined.RESULTS: The preparation assumed as yellow transparent solution.The linear range of ethacridine was 5～50 ?g?mL-1(r=0.999 9) with an average recovery of 101.47% (RSD=1.32%,n=9).The content of ethacridine in the solution was not changed after sterilization.CONCLUSION: The quality of prepared ethacridine lactate solution is up to the standard.