Effectiveness and Safety of Apatinib Plus Programmed Cell Death Protein 1 Blockades for Patients with Treatment-refractory Metastatic Colorectal Cancer:A Retrospective Exploratory Study

This study aimed to investigate the efficacy and safety of apatinib plus programmed cell death protein 1 (PD-1) blockades for patients with metastatic colorectal cancer (CRC) who were refractory to the standard regimens. In this retrospective study, patients with metastatic CRC who received apatinib plus PD-1 blockades in clinical practice were included. The initial dosage of apatinib was 250 mg or 500 mg, and PD-1 blockades were comprised of camrelizumab, sintilimab and pembrolizumab. Efficacy and safety data were collected through the hospital’s electronic medical record system. From October 2018 to March 2022, a total of 43 patients with metastatic CRC were evaluated for efficacy and safety. The results showed an objective response rate of 25.6% (95% CI, 13.5%-41.2%) and a disease control rate of 72.1% (95% CI, 56.3%-84.7%). The median progression-free survival (PFS) of the cohort was 5.8 months (95% CI, 3.81-7.79), and the median overall survival (OS) was 10.3 months (95% CI, 5.75-14.85). The most common adverse reactions were fatigue (76.7%), hypertension (72.1%), diarrhea (62.8%), and hand-foot syndrome (51.2%).Multivariate Cox regression analysis revealed that Eastern Cooperative Oncology Group (ECOG) performance status and location of CRC (left or right-side) were independent factors to predict PFS of patients with metastatic CRC treated with the combination regimen. Consequently, the combination of apatinib and PD-1 blockades demonstrated potential efficacy and acceptable safety for patients with treatment-refractory metastatic CRC. This conclusion should be confirmed in prospective clinical trials subsequently.

Neoadjuvant PD-1 Inhibitor Plus Apatinib and Chemotherapy Versus Apatinib Plus Chemotherapy in Treating Patients With Locally Advanced Gastric Cancer: A Prospective, Cohort Study

Purpose@#This study aimed to evaluate the efficacy and safety of neoadjuvant programmed cell death-1 (PD-1) inhibitors plus apatinib and chemotherapy (PAC) in patients with locally advanced gastric cancer (LAGC). @*Materials and Methods@#Seventy-three patients with resectable LAGC were enrolled and named the PAC group (n=39) or apatinib plus chemotherapy (AC) group (n=34) based on the treatment they chose. Neoadjuvant therapy was administered in a 21-day cycle for 3 consecutive cycles, after which surgery was performed. @*Results@#The PAC group exhibited a higher objective response rate than the AC group (74.4% vs. 58.8%, P=0.159). Moreover, the PAC group showed a numerically better response profile than the AC group (P=0.081). Strikingly, progression-free survival (PFS) (P=0.019) and overall survival (OS) (P=0.049) were prolonged, whereas disease-free survival (DFS) tended to be longer in the PAC group than in the AC group (P=0.056). Briefly, the 3-year PFS, DFS, and OS rates were 76.1%, 76.1%, and 86.7% in the PAC group and 46.9%, 49.9%, and 70.3% in the AC group, respectively. Furthermore, PAC (vs. AC) treatment (hazard ratio=0.286, P=0.034) was independently associated with prolonged PFS in multivariate Cox regression analyses. The incidence of adverse events did not differ between the two groups (all P>0.05), where leukopenia, anemia, hypertension, and other adverse events were commonly observed in the PAC group. @*Conclusions@#Neoadjuvant PAC therapy may achieve a preferable pathological response, delayed progression, and prolonged survival compared to AC therapy with a similar safety profile in patients with LAGC; however, further validation is warranted.

Establishment and characterization of bone marrow mesenchymal stem cell lines stably synthesizing high-level dopamine / 生物工程学报

A triple-transgenic (tyrosine hydroxylase/dopamine decarboxylase/GTP cyclohydrolase 1, TH/DDC/GCH1) bone marrow mesenchymal stem cell line (BMSCs) capable of stably synthesizing dopamine (DA) transmitters were established to provide experimental evidence for the clinical treatment of Parkinson's disease (PD) by using this cell line. The DA-BMSCs cell line that could stably synthesize and secrete DA transmitters was established by using the triple transgenic recombinant lentivirus. The triple transgenes (TH/DDC/GCH1) expression in DA-BMSCs was detected using reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immunofluorescence. Moreover, the secretion of DA was tested by enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC). Chromosome G-banding analysis was used to detect the genetic stability of DA-BMSCs. Subsequently, the DA-BMSCs were stereotactically transplanted into the right medial forebrain bundle (MFB) of Parkinson's rat models to detect their survival and differentiation in the intracerebral microenvironment of PD rats. Apomorphine (APO)-induced rotation test was used to detect the improvement of motor dysfunction in PD rat models with cell transplantation. The TH, DDC and GCH1 were expressed stably and efficiently in the DA-BMSCs cell line, but not expressed in the normal rat BMSCs. The concentration of DA in the cell culture supernatant of the triple transgenic group (DA-BMSCs) and the LV-TH group was extremely significantly higher than that of the standard BMSCs control group (P < 0.000 1). After passage, DA-BMSCs stably produced DA. Karyotype G-banding analysis showed that the vast majority of DA-BMSCs maintained normal diploid karyotypes (94.5%). Moreover, after 4 weeks of transplantation into the brain of PD rats, DA-BMSCs significantly improved the movement disorder of PD rat models, survived in a large amount in the brain microenvironment, differentiated into TH-positive and GFAP-positive cells, and upregulated the DA level in the injured area of the brain. The triple-transgenic DA-BMSCs cell line that stably produced DA, survived in large numbers, and differentiated in the rat brain was successfully established, laying a foundation for the treatment of PD using engineered culture and transplantation of DA-BMSCs.

Risk factors and prognosis of varicella zoster virus infection following hematopoietic stem cell transplantation in children with thalassemia / 中华实用儿科临床杂志

Objective:To investigate the risk factors and prognosis of varicella zoster virus (VZV) infection in children with thalassemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The clinical data of 446 children with thalassemia who underwent allo-HSCT from January 2012 to December 2020 in the Department of Hematology and Oncology, Shenzhen Children′s Hospital were retrospectively collected.The clinical features of the patients with VZV infection were analyzed.The patients were divided into different groups according to whether they had VZV infection.Categorical variables between groups were compared using the chi- square tests to investigate the risk factors that were associated with the development of VZV.Survival time was analyzed by the Kaplan-Meier method. Results:VZV incidence was 4.3% (19/446 cases), and the median onset time was 5 months (1.5-11.0 months) after allo-HSCT.Of the 19 cases with VZV infection, 5 cases were complicated with VZV encephalitis.All cases were treated with antiviral agents (Acyclovir alone, or both Acyclovir and Foscarnet), intravenous immunoglobulin and external use of Acyclovir ointment.After 7-28 days of treatment (median treatment time: 14 days), all of their herpes subsided, and the neurological symptoms of patients with VZV encephalitis disappeared.One of the 19 children died.The death was not directly caused by VZV infection, but by secondary graft dysfunction and severe pneumonia 5 months after VZV infection.The incidence of VZV infection following allo-HSCT in children with thalassemia was related to the age of the donor ( P=0.010), but not to the age of the patient ( P=0.378), gender ( P=0.653), disease grade of thalassemia ( P=0.912), type of the donor ( P=0.205), source of stem cells ( P=0.624) and acute graft versus host disease ( P=0.277). VZV infection had no significant effect on the prognosis of thalassemia children after allo-HSCT ( P=0.241). Conclusions:Thalassemia children with VZV infection after allo-HSCT are prone to be complicated with VZV encephalitis.Cord blood transplantation is a high risk factor.VZV infection may not have an impact on survival of children with thalassemia after allo-HSCT.

Risk factors and prognosis of varicella zoster virus infection following hematopoietic stem cell transplantation in children with thalassemia / 中华实用儿科临床杂志

Objective:To investigate the risk factors and prognosis of varicella zoster virus (VZV) infection in children with thalassemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The clinical data of 446 children with thalassemia who underwent allo-HSCT from January 2012 to December 2020 in the Department of Hematology and Oncology, Shenzhen Children′s Hospital were retrospectively collected.The clinical features of the patients with VZV infection were analyzed.The patients were divided into different groups according to whether they had VZV infection.Categorical variables between groups were compared using the chi- square tests to investigate the risk factors that were associated with the development of VZV.Survival time was analyzed by the Kaplan-Meier method. Results:VZV incidence was 4.3% (19/446 cases), and the median onset time was 5 months (1.5-11.0 months) after allo-HSCT.Of the 19 cases with VZV infection, 5 cases were complicated with VZV encephalitis.All cases were treated with antiviral agents (Acyclovir alone, or both Acyclovir and Foscarnet), intravenous immunoglobulin and external use of Acyclovir ointment.After 7-28 days of treatment (median treatment time: 14 days), all of their herpes subsided, and the neurological symptoms of patients with VZV encephalitis disappeared.One of the 19 children died.The death was not directly caused by VZV infection, but by secondary graft dysfunction and severe pneumonia 5 months after VZV infection.The incidence of VZV infection following allo-HSCT in children with thalassemia was related to the age of the donor ( P=0.010), but not to the age of the patient ( P=0.378), gender ( P=0.653), disease grade of thalassemia ( P=0.912), type of the donor ( P=0.205), source of stem cells ( P=0.624) and acute graft versus host disease ( P=0.277). VZV infection had no significant effect on the prognosis of thalassemia children after allo-HSCT ( P=0.241). Conclusions:Thalassemia children with VZV infection after allo-HSCT are prone to be complicated with VZV encephalitis.Cord blood transplantation is a high risk factor.VZV infection may not have an impact on survival of children with thalassemia after allo-HSCT.

Expression of programmed death-1 and programmed death-ligand 1 in post-transplant lymphoproliferative disorders after allogeneic hematopoietic stem cell transplantation in children / 中华实用儿科临床杂志

Objective:To explore the expressions of programmed death-1 (PD-1) and programmed death-ligand 1 (PD-L1) and clinicopathological characteristics in post-transplant lymphoproliferative disorder (PTLD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children, with the aim of clarifying whether checkpoint inhibition of PD-1/PD-L1 inhibitors may serve as a therapy option.Methods:The clinical data of 13 cases of PTLD after allo-HSCT pathologically confirmed in Shenzhen Children′s Hospital from January 1, 2012 to December 30, 2019 were retrospectively analyzed.The detection was performed by immunohistochemical staining by MaxVision? method, Epstein-Barr virus(EBV) in situ hybridization and lymphoma gene rearrangement.The relationship between the expression of PD-1 and PD-L1 and the clinicopathological characteristics of PTLD were analyzed.Results:The expression of PD-1 was not correlated with gender, age, primary diseases, histopathological types, transplantation mode and the expression of EBV in situ hybridization (all P>0.05). The expression of PD-L1 was correlated with histopathological types ( P<0.05). Furthermore, the expression rate of PD-L1 on severe β-thalassemia was significantly higher than that of severe aplastic anemia [90.0%(9/10 cases) vs. 66.7%(2/3 cases)] and monomorphic PTLD was higher than that of polymorphic PTLD [100.0%(2/2 cases) vs. 83.3%(5/6 cases)]. Moreover, the positive PTLD in EBV was higher than the negative PTLD in EBV [90.9%(10/11 cases) vs. 50.0%(1/2 cases)]. The positive rates of PD-1 and PD-L1 in 13 cases with PTLD were 46.2%(6/13 cases) and 61.5%(8/13 cases) in tumor cells, 92.3% (12/13 cases) and 76.9% (10/13 cases) in microenvironmental cells, and 84.6%(11/13 cases) in EBV, respectively. Conclusions:PD-L1 has a higher positive rate in tumor cells with monomorphic PTLD; and routine staining for PD-1 and PD-L1 can be performed in all types of PTLD when standard immunotherapy and chemotherapy are ineffective.

Clinical and genetic analysis of a child with transcobalamin II deficiency / 中华医学遗传学杂志

OBJECTIVE@#To investigate the genetic etiology, clinical diagnosis and treatment of a child with pancytopenia, failure to thrive and pulmonary infection.@*METHODS@#Peripheral blood samples of the child and her parents were collected. Genomic DNA was extracted. Genetic variants associated with hematological diseases were detected by high-throughput sequencing.@*RESULTS@#Three variants of TCN2 gene were found, one of which located in exon 5 upstream(c.581-8A>T), the parents has carried this variant; one in exon 6 (c.924_927del), the variant was originated from the mother; one in exon 7 (c.973C>T), the variant has ocurred de novo. The variants pathogenic analysis combined with clinical manifestation, pancytopenia, the increase in methylmalonic acid level and increased homocysteine, the child was diagnosed with transcobalaminIIdeficiency. The patient presented with respiratory infection, which was confirmed to be pneumocystosis by lung radioscopy and pathogenic high-throughput sequencing of broncho-alveolar lavage fluid. The patient presented with acute respiratory distress syndrome during the treatment with intramuscular injection of vitamin B@*CONCLUSION@#We reported a case of Chinese child with TCNII deficiency due to novel gene variant, and analyzed the pathogenicity of the three variants. The treatment of TCNII deficiency with cobalamin should be individualized.

Relationship between autophagy regulated by chromosome 19 open reading frame 5 and malignant degree of colorectal cancer and chemosensitivity of paclitaxe / 中国医师进修杂志

Objective:To observe the relationship between autophagy regulation activity of chromosome 19 open reading frame 5 (C19ORF5) and malignant degree of colorectal cancer and chemosensitivity of paclitaxel.Methods:The tumor tissues and normal adjacent tissues of 141 colorectal cancer patients from 2015 to 2017 in the Second Affiliated Hospital of Harbin Medical University were selected. The expressions of C19ORF5 protein and mRNA were detected by immunohistochemistry and timed quantitative polymerase chain reaction. The correlation between C19ORF5 protein regulating autophagy activity and malignancy of colorectal cancer was analyzed. All 141 patients received postoperative chemotherapy, among whom 91 patients received conventional chemotherapy (capecitabine combined with oxaliplatin, conventional chemotherapy group), and 50 patients received conventional chemotherapy combined with paclitaxel (paclitaxel group). Six course of treatment was treated in both groups.Results:Autophagosomes could been seen under transmission electron microscopy. C19ORF5 protein was pale yellow to tan granules, and was expressed in the cytoplasm. The C19ORF5 protein staining intensity of cancer tissue was significantly stronger than that of normal tissue, and the staining intensity of stage Ⅱ was significantly higher than that of stage Ⅳ. The high expression rate of C19ORF5 protein in patients with stage Ⅰ to Ⅱ was significantly higher than that in patients with stage Ⅲ to Ⅳ: 83.3% (25/30) vs. 17.1% (19/111), and there was statistical difference ( P<0.01). The expression of C19ORF5 mRNA in cancer tissues was significantly higher than that in normal tissues: 1.17 ± 0.45 vs. 0.82 ± 0.29, and there was statistical difference ( P<0.05). The expression level of C19ORF5 protein in cancer tissue was related to tumor stage, carcinoembryonic antigen and liver metastasis ( P<0.01 or <0.05); the expression level of C19ORF5 protein in cancer tissue was not related to lymph node metastasis ( P>0.05). Of the 91 cases in conventional chemotherapy group, chemotherapy was effective in 70 cases (76.9%) and ineffective in 21 cases (23.1%). Of 50 cases in paclitaxel group, it was effective in 42 cases (84.0%) and ineffective in 8 cases (16.0%). There was no statistical difference in effective rate between 2 groups ( P>0.05). In conventional chemotherapy group, there was no significant difference in serum C19ORF5 protein expression levels between cancer tissues before and after chemotherapy in effective patients and ineffective patients ( P>0.05); there was no significant difference in serum C19ORF5 protein expression levels between effective patients and ineffective patients after chemotherapy ( P>0.05). In paclitaxel group, the expression level of C19ORF5 protein in cancer tissues before chemotherapy was significantly higher than that in serum C19ORF5 protein after chemotherapy: 0.9 ± 0.3 vs. 0.5 ± 0.2, the expression level of serum C19ORF5 protein in patients with effective chemotherapy was significantly lower than that in ineffective patients: 0.5 ± 0.2 vs. 0.8 ± 0.2, and there were statistical differences ( P<0.05). Conclusions:The high expression of C19ORF5 protein can increase the autophagy activity of colorectal cancer tissue; C19ORF5 protein regulates autophagy activity and is negatively correlated with the malignant degree of colorectal cancer; C19ORF5 protein may increase the sensitivity of paclitaxel chemotherapy by enhancing autophagy activity.

The comparison of effectiveness and safety of mechanical continue chest compression ventilation (CCV) and interrupt pressure ventilation (30∶2) on cardiopulmonary resuscitation / 中国医师杂志

Objective To investigate the effects and safety of mechanical continue heart chest compression ventilation (CCV) and interrupt pressure ventilation (30∶2) on cardiopulmonary resuscitation (CPR).Methods According to the mechanical pressure ventilation different way,90 patients with cardiopulmonary resuscitation (CPR) in our hospital emergency center were admitted into control group (30∶2 group) with 48 cases and observation group (CCV group) with 42 cases,with a before and after case-control study to compare the recovery effects [rate of spontaneous circulation restoration (ROCS),successful recovery rate,the ROCS time,withdraw machine time,and Glasgow coma scale(GCS) in successful recovery patients] and recovery of adveme events (fracture,skin lesions,hemopneumothorax,and visceral injury).Results With comparison between two groups,CCV group had shorter ROCS recovery time and withdraw machine time [(9.31±4.32) min vs (12.67±4.86) min,(32.07 t4.84) min vs (36.33 ± 3.37)min,P ＜0.05],higher rate of ROCS and successful recovery rate (42.9％ vs 22.9％,21.4％ vs 6.3％,P ＜ 0.05) than 30∶2 group;while both were no difference in Glasgow coma scale (GCS) in successful recovery patients (P ＞ 0.05).The incidence of adverse events were not different in both (11.9％ vs 8.3％,P ＞0.05).Conclusions Mechanical continue chest compression ventilation (CCV) compared to interrupt pressure ventilation (30∶ 2) can effectively shorten the ROCS recovery time and withdraw machine time,improve the ROSC recovery rate and success rate of recovery,but failed to improve nerve functions.Two groups have a lower incidence of adverse events,which is safe to use.

Expression of a recombinant protein by fusing anti-CD20 ScFv with dominant antigen epitopes and its anti-tumor activity / 中国免疫学杂志

Objective:To artificially design and express a recombinant protein named as ScFv-pLLO by fusing ScFv gene of Rituximab(C2B8)and dominant antigen epitopes from listeriolysin O(LLO),and studying its anti-tumor activity.Methods:VH and VL gene sequences of C2B8 against CD20 were acquired by searching United States Patent database,and ScFv sequence was constructed by linking VL and VH with a short peptide linker.Two CD4+T cell epitopes from LLO were selected and designed to splice ScFv sequence.The recombinant gene of ScFv-pLLO was cloned into prokaryotic expression vector and purified after induction.The capacity of ScFv-pLLO target-binding to B-cell lymphomas was evaluated by flow cytometry ( FCM ) and co-immunoprecipitation ( Co-IP ) .The effects of ScFv-pLLO on B-cell lymphomas proliferation and apoptosis were detected respectively.The immunogenicity of ScFv-pLLO was assessed by lymphocyte proliferation assay.Results: ScFv-pLLO was successfully expressed.It could bind to different B-cell lymphomas cell lines and obviously inhibit the growth of Raji cells as well as inducing apoptosis.Moreover,ScFv-pLLO was able to stimulate proliferation of spleen lymphocytes of immunized mice.Conclusion: The recombinant protein ScFv-pLLO can target-bind to B-cell lymphomas,and perform inhibitory effect and induce apoptosis on Raji cells that indicate ScFv-pLLO retain the capacity of ScFv derived from monoclonal antibody against CD20.Besides, ScFv-pLLO can induce immune response.This study provides a basis for further research about the role of ScFv-pLLO on simulating tumor cell antigens as well as being tumor vaccine adjuvant.

Culture and characterization of human fetal hepatocytes / 基础医学与临床

Objective Determining the characteristics of human fetal hepatocytes in vitro. Methods Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the secretion of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion Human fetal hepatocytes can be maintained keeping function in vitro for several months.

Species identification of animal cells by polymerase chain reaction / 基础医学与临床

Objective To identify the species of cultured cells and to detect inter-species cross contamination by polymerase chain reaction (PCR).Methods From references and NCBI database,we outsourced 32 pairs of species-specific primers targeted to genomic DNA of ten common species of cultured cells.Then we screened for optional primers with high specificity and high sensitivity.PCR amplification with these primers,the genomic DNAs isolated from the tested cell line and agarose gel electrophoresis of the PCR products were done.Mixed DNA of 10 species was used as positive-control template,and water as negative-control template.Results Ten pairs of species-specific and highly sensitive primers were selected.By PCR amplification with these primers and agarose gel electrophoresis,we may easily identify the origin of cell lines and find whether the tested cell lines are contaminated by cells of other species.Conclusion This PCR assay provides a simple,rapid,sensitive,and cost-effective method to identify cell species and detect interspecies cross-contamination.

Determination of Trace Lead in Fresh Marine Products by Fluorescence Spectrophotometry / 环境与健康杂志

0.05). Conclusion The method is simple,accurate,reliable,less-reagent used and little inference of other metal ions. It is suitable for rapid determination of lead in the fresh marine products.