GBA inhibition suppresses ovarian cancer growth, survival and receptor tyrosine kinase AXL-mediated signaling pathways

The poor outcome of advanced ovarian cancer under conventional therapy necessitates new strategies to improve therapeutic efficacy. β-glucosidase (encoded by GBA) is a lysosomal enzyme and is involved in sphingolipids metabolism. Recent studies revealed that β-glucosidase plays a role in cancer development and chemoresistance. In this work, we systematically evaluated the expression and role of GBA in ovarian cancer. Our work demonstrates that inhibition of β-glucosidase has therapeutic potential for ovarian cancer. Gene Expression Profiling Interactive Analysis database, western blot and immunohistochemistry analyses of patient samples demonstrated that GBA mRNA and protein expression levels were significantly increased in ovarian cancer compared to normal tissues. Functional studies using gainof-function and loss-of-function approaches demonstrated that GBA overexpression did not affect growth and migration but alleviated cisplatin’s efficacy in ovarian cancer cells. In addition, GBA depletion resulted in growth inhibition, apoptosis induction, and enhancement of cisplatin’s efficacy. Of note, we found that GBA inhibition specifically decreased receptor tyrosine kinase AXL level, leading to the suppression of AXL-mediated signaling pathways. Our data suggest that GBA represents a promising target to inhibit AXL signaling and overcome cisplatin resistance in ovarian cancer.

Analysis of clinical presentation and genetic characteristics of malignant infantile osteopetrosis / 中华儿科杂志

Objective: To investigate the clinical presentation and genetic characteristics of malignant infantile osteopetrosis. Methods: This was a retrospective case study. Thirty-seven children with malignant infantile osteopetrosis admitted into Beijing Children's Hospital from January 2013 to September 2022 were enrolled in this study. According to the gene mutations, the patients were divided into the CLCN7 group and the TCIRG1 group. Clinical characteristics, laboratory tests, and prognosis were compared between two groups. Wilcoxon test or Fisher exact test were used in inter-group comparison. The survival rate was estimated with the Kaplan-Meier method and the Log-Rank test was used to compare the difference in survival between groups. Results: Among the 37 cases, there were 22 males and 15 females. The age of diagnosis was 0.5 (0.2, 1.0) year. There were 13 patients (35%) and 24 patients (65%) with mutations in CLCN7 and TCIRGI gene respectively. Patients in the CLCN7 group had an older age of diagnosis than those in the TCIRGI group (1.2 (0.4, 3.6) vs. 0.4 (0.2, 0.6) years, Z=-2.60, P=0.008). The levels of serum phosphorus (1.7 (1.3, 1.8) vs. 1.1 (0.8, 1.6) mmol/L, Z=-2.59, P=0.010), creatine kinase isoenzyme (CK-MB) (457 (143, 610) vs. 56 (37, 82) U/L, Z=-3.38, P=0.001) and the level of neutrophils (14.0 (9.9, 18.1) vs. 9.2 (6.7, 11.1) ×109/L, Z=-2.07, P=0.039) at diagnosis were higher in the CLCN7 group than that in the TCIRG1 group. However, the level of D-dimer in the CLCN7 group was lower than that in the TCIRGI group (2.7 (1.0, 3.1) vs. 6.3 (2.5, 9.7) μg/L, Z=2.83, P=0.005). After hematopoietic stem cell transplantation, there was no significant difference in 5-year overall survival rate between the two groups (92.3%±7.4% vs. 83.3%±7.6%, χ²=0.56, P=0.456). Conclusions: TCIRGI gene mutations are more common in children with osteopetrosis. Children with TCIRGI gene mutations have younger age, lower levels of phosphorus, CK-MB, and neutrophils and higher level of D-dimer at the onset. After hematopoietic stem cell transplantation, patients with CLCN7 or TCIRGI gene mutations have similar prognosis.

Alterations in the intestinal microbiota of preterm infants with neurodevelopmental impairments: a prospective cohort study / 中国当代儿科杂志

OBJECTIVES@#To investigate the difference in intestinal microbiota between preterm infants with neurodevelopmental impairment (NDI) and those without NDI.@*METHODS@#In this prospective cohort study, the preterm infants who were admitted to the neonatal intensive care unit of Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from September 1, 2019 to September 30, 2021 were enrolled as subjects. According to the assessment results of Gesell Developmental Scale at the corrected gestational age of 1.5-2 years, they were divided into two groups: normal (n=115) and NDI (n=100). Fecal samples were collected one day before discharge, one day before introducing solid food, and at the corrected gestational age of 1 year. High-throughput sequencing was used to compare the composition of intestinal microbiota between groups.@*RESULTS@#Compared with the normal group, the NDI group had a significantly higher Shannon diversity index at the corrected gestational age of 1 year (P<0.05). The principal coordinate analysis showed a significant difference in the composition of intestinal microbiota between the two groups one day before introducing solid food and at the corrected gestational age of 1 year (P<0.05). Compared with the normal group, the NDI group had a significantly higher abundance of Bifidobacterium in the intestine at all three time points, a significantly higher abundance of Enterococcus one day before introducing solid food and at the corrected gestational age of 1 year, and a significantly lower abundance of Akkermansia one day before introducing solid food (P<0.05).@*CONCLUSIONS@#There are significant differences in the composition of intestinal microbiota between preterm infants with NDI and those without NDI. This study enriches the data on the characteristics of intestinal microbiota in preterm infants with NDI and provides reference for the microbiota therapy and intervention for NDI in preterm infants.

Joint association of sedentary behavior and physical activity on anxiety tendency among occupational population in China / 北京大学学报(医学版)

OBJECTIVE@#To investigate the status and influencing factors of anxiety tendency among occupational population in China and to examine the joint association between sedentary behavior and physical activity with anxiety tendency.@*METHODS@#The data were from the 2021 Asia Best Workplace (Chinese mainland) program. The Generalized Anxiety Tendency scale was used to assess employees' anxiety status, and Logistic regression was used to analyze the factors influencing anxiety tendency and calculate the odds ratio (OR) within different groups. The OR of sitting for each sitting-physical activity (PA) combination group and within PA strata were calculated to explore the joint association.@*RESULTS@#A total of 11 903 workers with an average age of 32.9 years were included in this study. Among them, 3 562 workers had anxiety tendency (29.9%) and the prevalence of those under 40 years old (30.6%) was significantly higher than the other age group (26.7%). 41.0% of the respondents had the moderated to vigorous physical activity. Their average daily sitting time was 9.4 h, and the percentage of those who exceeded 8 h sitting reached 73.9% in the past week. The analysis of Logistic regression showed that smoking (OR=1.24, 95%CI: 1.23-1.39), longer sedentary time and lower physical activity level were risk factors for anxiety tendency, and longer average daily sleep time (OR=0.56, 95%CI: 0.51-0.61) was a protective factor. The joint association analysis and stratified analysis of physical activity and sedentary behavior with anxiety tendency showed that increased sedentary time combined with decreased physical activity intensity was significantly associated with increased risk of anxiety tendency (range of OR: 1.64-3.14). The threshold for sedentary time in total as a risk factor for anxiety tendency gradually decreased as physical activity intensity increased.@*CONCLUSION@#The anxiety tendency and sedentary behavior among the occupational population should recieve more attention. Lack of physical activity and sedentary behavior are both risk factors for anxiety tendency, and strengthening the intensity of physical activity can attenuate the harmful effects of sedentary behavior on anxiety tendency.

Host-schistosome interactions mediated by cross-species microRNA / 中国热带医学

@#Abstract: Schistosomiasis is a serious major parasitic disease that threatens human life and health. A better understanding of the mechanism of host-schistosome interactions is the key to designing new prevention and control strategies. MicroRNAs (miRNAs) are endogenous small non-coding RNA molecules, which lead to the degradation of the target messenger RNA (mRNA) or inhibition of its translation in a sequence-specific manner. Both schistosome and its host produce miRNAs, which can be secreted by extracellular vesicles (EVs). There is accumulating evidence that miRNAs from schistosome can be taken up by host cells, and finely manipulate the phenotype of host cells for their survival or pathogenesis in a cross-species manner, even inhibiting the growth and metastases of hepatoma cells. It is still unknown whether host free miRNAs can be taken up by schistosome, but this phenomenon is highly probable. miRNA-mediated cross-species regulation has emerged as a novel mechanism for host-schistosome interactions, and this review summarizes the advances in this regard.

Effect of Pelvic Floor Muscle Training on Neurogenic Detrusor Overactivity / 中国康复理论与实践

Objective:To investigate the effect of pelvic floor muscle training on detrusor overactivity in patients with spinal cord injury. Methods:From August, 2019 to August, 2020, 17 patients with incomplete spinal cord injury were randomly divided into control group (n = 8) and experimental group (n = 9). Both groups accepted tolterodine tartrate 4 mg a day orally for twelve weeks, while the experimental group accepted pelvic floor muscle training. They were measured urodynamic indexes, and assessed with Neurogenic Bladder Symptom Scale and Patient Perception of Bladder Condition before and after treatment. Results:All the urodynamic indexes and scores of the scales improved in both groups after treatment (t > 3.674, |Z| > 2.646, P < 0.05), while the indexes of maximum bladder volume, first contraction pressure volume of detrusor, leakage point pressure of detrusor, maximum contraction pressure of detrusor and neurogenic bladder symptom score improved more in the experimental group than in the control group (|t| > 2.194, P < 0.05). Conclusion:Pelvic floor muscle training based on medicine can release the detrusor overactivity in patients with spinal cord injury.

Effects of atmospheric particulates exposure on Nrf2 signaling pathway and inflammatory factors in myocardium of rats fed with high-fat and high-glucose diet / 上海预防医学

Objective To study the effect of atmospheric particulate exposure on the expression of key molecules of Nrf2 signaling pathway involved in oxidative stress and inflammatory response factors in myocardium of rats fed with high-fat and high-glucose diet. Methods A total of 48 SD male rats were randomly divided into control group (CC group), high-fat and high-glucose diet group (HC group), atmospheric particulates group (CP group) and atmospheric particulates plus high-fat and high-glucose diet group (HP group), with 12 rats in each group.Rats were fed in individual ventilated cages (IVC).The CC and HC groups were placed in IVCs equipped with the atmospheric particulate filter, however, the CP and HP groups without the atmospheric particulate filter to make the air composition similar to the outdoor.A total of 24 rats were sacrificed for acquiring myocardial tissue after 3 and 6 months of exposure.The mRNA expression of Nrf2, HO-1, NQO1, VCAM-1 and MCP-1 were measured using RT-qPCR and the protein expression of VCAM-1, MCP-1 detected using western blot. Results The mRNA expression levels of Nrf2, HO-1, NQO1, VCAM-1 and MCP-1 and the protein expression levels of VCAM-1 and MCP-1 in HC, CP and HP groups were higher than CC group (P < 0.05).The mRNA expression levels of Nrf2, HO-1, NQO1, VCAM-1, MCP-1 and the protein expression levels of VCAM-1, MCP-1 in the HP group were higher than HC and CP groups (P < 0.05).The mRNA expression levels of Nrf2 in CP and HP groups after 6 months of exposure were lower than that at 3 months (P < 0.05). Conclusion The exposure of atmospheric particles, high-fat and high-glucose and their combination diets could cause myocardial tissue inflammatory responses, and activate Nrf2 signaling pathways to protect against myocardial damage.

A clinical study of haploidentical hematopoietic stem cell transplantation in the treatment of pediatric patients with acquired severe aplastic anemia: single center experience / 中华血液学杂志

Objective: To investigate the efficacy of haplotype hematopoietic stem cell transplantation in the treatment of acquired severe aplastic anemia (SAA) in children. Methods: The clinical characteristics of 59 pediatric patients with SAA, including 26 cases VSAA, 37males and 22 females, 47 cases typeⅠ and 12 cases typeⅡ, undrerwent haplo-HSCT in our hospital between December 1st, 2011 and December 1st, 2017 were retrospectively analyzed. Among 59 patients, 56 patients with a median age of 4.5 (1.2-14.8) years and median weight of 43 (12-80) kg underwent their first HSCT and 3 patients underwent their second HSCT. All patients received the following conditioning regimen: busulfan, cyclophosphamide, and rabbit ATG or Bu (-, CTX) , fludarabineand rabbit ATG. The prophylaxis of acute graft versus host disease (aGVHD) was cyclosporine (CsA) , MMF and methotrexate. All patients received bone marrow transfusion on day 01 and peripheral stem cell transfusion on day 02 from haploid donor. The median dose of donor mononuclear cell counts was 15.60 (7.74-21.04) ×108/kg of recipient weight and CD34+ cell counts was 4.86 (3.74-7.14) ×106/kg of recipient weight. Results: Neutrophils and platelets of all 59 children were implanted. The median implantation time of granulocytes and platelets were 13 (10-19) d, 19 (9-62) d, respectively. The incidence of grade Ⅰ-Ⅱ aGVHD was 45.76% (27 cases) and grade Ⅲ/Ⅳ 13.56% (8 cases) , The incidence of chronic GVHD was 8.47% (5 cases) , The incidences of CMV and EBV viremia were 59.32% (35 cases) and 28.81% (17 cases) , respectively. The median follow-up was 30 (8-80) months, 57 patients survived with disease free, 2 patients died of GVHD. Both of the estimated 5-year OS and DFS rates were (96.4±2.5) %. Conclusion: Haplo-HSCT could improve the outcomes of SAA children.

A clinical study of haploid hematopoietic stem cells combined with third-party umbilical cord blood transplantation in the treatment of chronic granulomatous disease / 中国当代儿科杂志

OBJECTIVE@#To investigate the clinical efficacy of haploid hematopoietic stem cells (haplo-HSC) combined with third-party umbilical cord blood (tpCB) transplantation in the treatment of X-linked chronic granulomatous disease (X-CGD).@*METHODS@#The clinical data of 26 boys with X-CGD were retrospectively analyzed who were admitted to the Sixth Medical Center of PLA General Hospital between April 2014 and March 2018. All the patients were treated with haplo-HSC combined with tpCB transplantation. The median age of the patients was 3.5 years. The donor was the father in 25 cases and an aunt in 1 case. Transplantation was 5/6 HLA-matched in 9 cases, 4/6 in 12 cases, and 3/6 in 5 cases. The patients received busulfan, cyclophosphamide, fludarabine, or anti-thymocyte globulin for myeloablative preconditioning. Cyclosporine A and mycophenolate mofetil were used for prevention of acute graft-versus-host disease (aGVHD). Then the patients were treated with haploid bone marrow hematopoietic stem cells combined with tpCB transplantation on day 1 and haploid peripheral hematopoietic stem cells on day 2. The counts of median donor total nucleated cells, CD34 cells, and CD3 cells were 14.6×10/kg, 5.86×10/kg, and 2.13×10/kg respectively.@*RESULTS@#The median time to neutrophil and platelet engraftment was 12 and 23 days after transplantation respectively. Full donor hematopoietic chimerism was observed on day 30. Twenty-five cases were from haplo-HSC and 1 was from cord blood. No primary implant failure and implant dysfunction occurred, and secondary implant failure occurred in one case. The NADPH oxidase activity returned to normal one month after transplantation. The incidence of grade I-II aGVHD and grade III-IV aGVHD was 35% and 15% respectively. Chronic GVHD (cGVHD) of the skin occurred in one case, and no progression was observed after steroid administration. During the follow-up period of 6-51 months, 25 patients survived, of whom 24 were disease-free (23 patients without cGVHD and 1 with cGVHD of the skin) and NADPH oxidase activity returned to normal; one patient developed secondary implant failure but survived; one patient died of viral interstitial pneumonia 16 months after transplantation. The 5-year event-free survival rate and overall survival rate were 81%±12% and 89%±10% respectively.@*CONCLUSIONS@#Haplo-HSC combined with tpCB transplantation is one of the effective methods for the treatment of X-CGD in children.

Recent advances in the antitumor effects of evodiamine / 上海交通大学学报（医学版）

Evodiamine is the natural component extracted from Euodiae Fructus.Recently,growing evidence has proved that evodiamine has great effects on suppressing cell viability and proliferation,arresting cell cycle,inducing apoptosis,promoting autophagy,inhibiting the formation of microvascular angiogenesis as well as affecting epigenetic modification in cancer.Recent studies have continuously revealed related signal pathways involved in evodiamine such as PI3K-Akt and JAK-STAT pathways,as well as the impact of evodiamine on survivin,vascular endothelial growth factor and miRNAs.With the development and synthesis of evodiamine derivatives and related herbal formulations,the understanding of antitumor activity of evodiamine is gradually deepening.The important clinical significance and market value of evodiamine can be prospected.

Tumor derived IgG suppress the proliferation of T cells in cord blood / 北京大学学报(医学版)

Objective:To explore the function of tumor derived IgG (tIgG) and whether the tIgG can inhibit T cells activity.Methods:The tIgG was purified from ovarian cancer tissue.The cord blood monocyte cells (CBMC) and cord blood lymphocyte (CBL) were isolate from human umbilical cord blood.The CBMC and CBL were stimulated with phytohaemagg lutinin (PHA) in order to let the CBMC and CBL in the state of proliferation.Carboxyfluorescein succinimidyl amino ester (CFSE) was cultured with CBMC and CBL.CFSE had no cell toxicity,which could penetrate through the cell membrane and combine the intracellular protein.The fluorescence intensity decreased with the proliferation of cells step by step,so the proliferation of these cells could be detected in flow ctytometry.The tlgG which was puri fied from ovarian cancer tissue was divided into three groups,1 mg/L group,10 mg/L group,and 100 mg/L group,and the intravenous immunoglobulin (IVIG) was also divided into three groups too.The CBMC and CBL were treated by tIgG with 1 mg/L,10 mg/L,and 100 mg/L in order to observe the proliferation of T cells.The cells were treated with IVIG as a positive control group,and the cells were treated with phosphate buffer saline (PBS) as a negative control.The proliferation of CD4 + or CD8 + T cells were detected in CBMC and CBL.The proliferation of the T cells in CBMC and CBL after 64 h and 86 h were detected.Results:In the system of CBMC,the tIgG could suppress the proliferation of CD4 + or CD8 + T cells.The results could also be found in the system of CBL.The CD4 + or CD8 + T cells in the group which were treated with PBS were more active than those in the group which were treated with tIgG and IVIG.The suppression in the group which were treated with tIgG,was stronger than that in the group treated with IVIG.In addition,the suppression of T cells in the group which were stimulated with tIgG as 100 mg/L was more effective than that in the group which were stimulated with tIgG as 10 mg/L.This could prove that tIgG had the function of immunomodulation.Conclusion:The tIgG can be involved in immune escape of cancer.

MicroRNA expression based tumor origin classification / 临床检验杂志

Approximately 3 to 5％ of newly diagnosed metastatic cancers are of unknown primary tissue origin due to difficulties identifying a primary tumor using standard diagnostic approaches.MicroRNAs (miRNAs) have recently been demonstrated to be able to assist pathologist with improved accuracy in diagnosing cancers of unknown primary origin (CUP).In this short commentary,we will highlight some of the recent advancements in miRNA based cancer diagnosis as well as some future directions for the field.

Study on the spatial expression of trophectoderm cells in human embryonic prenatal blastocysts / 北京大学学报(医学版)

Objective:To study the spatial expression of trophectoderm cells in human embryonic preantral blastocysts.Methods:The study used Gardner score 5AA blastocysts harvested on day 6 after fertilization from assisted reproductive technology.Microcapsules were used to separate trophectoderm cells from the epidermal cells.Single-cell sequencing was performed.P ＜ 0.05 was calculated by unpaired t test,and the difference was 2 times.Here we determined,for the first time,global gene expression patterns in the polar/mural trophectoderm isolated from human blastocysts.Unsupervised hierarchical clustering analysis and gene ontology (GO) functional classification were performed using bioinformatics software.Differentially expressed genes were annotated by the Database for Annotation,Visualization and Integrated Discovery.Functions of differentially expressed genes were further annotated using encyclopedia of genes and genomes.Results:The results showed that there were up to 306 genes in the trophoblast cells and up to 75 genes in the trophoblast cells.Unsupervised cluster analysis of polar trophoblast cells and mural trophoblast cells were divided into two groups,belonging to different types and biological functions.Differences in gene function indicated that the biological functions of GO gene uptake genes were mainly transcription,energy metabolism,protein synthesis,transport,oxidative stress,ion transport,protein synthesis and transport,cell cycle regulation,actin growth,etc.They were mainly involved in ubiquitin-mediated protein hydrolysis,oxidative phosphorylation,Wnt signaling pathway,estrogen androgen metabolism and other signal pathways;wall trophoblast cells up-regulated gene GO biological function,which was mainly proteolytic metabolism,cell cycle arrest,apoptosis,activation of MAPK,carbohydrate transport,synaptic regulation,cell growth,calcium channel activation,positive B cell differentiation,T cell apoptosis and other biological functions,which were mainly involved in B cell receptor,T cell receptor,white blood cells cross-endothelial transplantation,VEGF expression,gap connection,GnRH secretion,apoptosis and other signaling pathways.Conclusion:The gene expression of blastocysts trophectoderm is revealed from the spatial dimension,indicating that differentiation of polar and mural trophectoderm of blastocysts is accompanied by differences between the two cell lineages,and the polar and mural trophectoderms are coordinated with each other and the blastocyst hatching and embryo implantation processes are finely adjusted.Further data analysis is expected to find the endogenous molecular specificity of the regulation of embryo implantation.

Identification and quantification of sesquiterpenes and polyacetylenes in Atractylodes lancea from various geographical origins using GC-MS analysis

A convenient and sensitive GC-MS method was developed to identify thirteen sesquiterpenes and polyacetylenes (e.g. caryophyllene, γ-elemene, α-caryophyllene, β-selinene, isoledene, germacrene B, elixene, atractylone, hinesol, β-eudesmol, atrctylodin, atractylenolide II and acetylatractylodinol) in Atractylodes lancea (Thunb.) DC., Asteraceae. Among those compounds, four major components including atractylone, hinesol, β-eudesmol and atrctylodin were quantified with standards; contents of other components were estimated by using calibration curve of hinesol. In this study, we presented that the concentrations of those thirteen components varied drastically in A. lancea samples from different producing areas. Among those components, atractylenolide II and acetylatractylodinol were identified by GC-MS for the first time. A hierarchical clustering analysis based on relative peak areas of those thirteen components in total ion current (TIC) profiles was used to characterize A. lancea samples from different producing areas. Further clustering analysis showed that a simplified method with only four major bioactive components could be used to serve the same aim.

Repressing malic enzyme 1 redirects glucose metabolism, unbalances the redox state, and attenuates migratory and invasive abilities in nasopharyngeal carcinoma cell lines / 癌症

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.

Clinically controlled study on children's infectious mononucleosis treated by Chinese medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy and safety of Chinese drugs for the treatment of children's infectious mononucleosis (CIM).</p><p><b>METHODS</b>Sixty CIM patients were assigned into the treated group and the control group, patients in the treated group were administered with Chinese herbal decoction, and those in the control group were treated with intravenous dripping of ganciclovir 10 mg/kg per day, for a treatment course of 14 days.</p><p><b>RESULTS</b>The total effective rate was 96.0% in the treated group and 97.1% in the control group, showing insignificant difference between groups. The efficacy in the treated group was superior to that in the control group on the fever clearance time (3.0+/-1.5 days vs 4.9+/-3.9 days ) and the disappearance time of cervical lymph node swelling (0.8+/-1.0 score vs 1.5+/-1.2 score), showing statistical significance (all P<0.05). T-cell subsets were markedly improved in both groups after treatment. Adverse reaction occurred in four cases of the control group.</p><p><b>CONCLUSION</b>Using Chinese herbs for clearing heat, removing toxin, activating blood circulation, and dissolving stasis is effective and safe for the treatment of CIM. It can effectively improve the clinical symptoms and shows a certain effect on immune regulation.</p>

Effect of Extracorporeal Shock Wave on Proliferating Activity of Human Mesenchymal Stem Cells / 中国康复理论与实践

@#ObjectiveTo explore the mechanism of extracorporeal shock wave (ESW) in treating osteogenic disorders and the ideal energy level. MethodsAfter success in marrow aspiration from patients' iliac crest, hMSCs were isolated by Percoll density gradient centrifugation and cultured in Dulbecco's modified Eagle's medium in a 5% CO2 and 37 ℃ incubator. Optimal ESW dose was determined by MTT of kinase-marked cytobiology. After hMSCs were exposed to ESW, their morphocytologic change, rate of adherence and doubling time were observed with IPCM. Enzyme cytochemistry reaction for the activity of alkaline phosphatase was also examined. ResultsESW of 5 kV and 100 times could increase cells' viability and proliferation (P<0.01), but higher than 7 kV would inhibit them. Rate of adherence of hMSCs in exposure group of passage 5 reached to 61.54%, which was significantly different from control group(P<0.05). Compared with control group, the MSCs' doubling time was short for 1.72 d (P<0.05). The curve of normal alkaline phosphatase activity of hMSCs was like type S, but ESW shortened its latent period, and promoted its peak time, which was significantly different from control group.ConclusionESW of 5 kV and 100 times can optimally promote the proliferation and activity of osteogen of hMSCs in vitro.

BPI700-Fc gamma1(700) chimeric gene expression and its protective effect in a mice model of the lethal E. coli infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Infections caused by gram-negative bacteria (GNB) often lead to high mortality in common clinical settings. The effect of traditional antibiotic therapy is hindered by drug-resistant bacteria and unneutralizable endotoxin. Few effective methods can protect high risk patients from bacterial infection. This study explored the protection of adeno-associated virus 2 (AAV2)-bacteriacidal permeability increasing protein 700 (BPI(700))-fragment crystallizable gamma one 700 (Fc gamma1(700)) chimeric gene transferred mice against the minimal lethal dose (MLD) of E. coli and application of gene therapy for bacterial infection.</p><p><b>METHODS</b>After AAV2-BPI(700)-Fc gamma1(700) virus transfection, dot blotting and Western blotting were used to detect the target gene products in Chinese hamster ovary-K1 cells (CHO-K1cells). Reverse transcription-polymerase chain reaction and immunohistochemical assay were carried out to show the target gene expression in mice. Modified BPI-enzyme linked immunosorbent assay was used to identify the target gene products in murine serum. The protection of BPI(700)-Fc gamma1(700) gene transferred mice was examined by survival rate after MLD E. coli challenge. Colony forming unit (CFU) count, limulus amebocyte lysate kit and cytokine kit were used to quantify the bacteria, the level of endotoxin, and proinflammatory cytokine.</p><p><b>RESULTS</b>BPI(1-199)-Fc gamma1 protein was identified in the CHO-K1 cell culture supernatant, injected muscles and serum of the gene transferred mice. After MLD E. coli challenge, the survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice (36.7%) was significantly higher than that of AAV2-enhanced green fluorescent protein (AAV2-EGFP) gene transferred mice (3.3%) and PBS control mice (5.6%). The survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice treated with cefuroxime sodium was 65.0%. The bacterium number in main viscera, the levels of endotoxin and proinflammatory cytokine (tumor necrosis factor-alpha and interleukin-1beta) in serum of the AAV2-BPI(700)-Fc gamma1(700) gene transferred mice were markedly lower than that of PBS control mice (P < 0.01).</p><p><b>CONCLUSIONS</b>AAV2-BPI(700)-Fc gamma1(700) gene transferred mice can resist MLD E. coli infection through expressing BPI(1-199)-Fc gamma1 protein. Our findings suggested that AAV2 mediated BPI(700)-Fc gamma1(700) gene delivery could be used for protection and treatment of clinical GNB infection in high-risk individuals.</p>