Increased tumor-associated mast cells facilitate thyroid cancer progression by inhibiting CD8+ T cell function through galectin-9

As an important component of solid tumors, mast cells show specific phenotypes in various tumor microenvironments. However, the precise mechanism of mast cell accumulation and the phenotypic features of thyroid cancer (TC) remain largely unknown. Here, we found that mast cells were obviously recruited to tumor tissue by TC-derived stem cell factor (SCF). With tumor progression, mast cell levels increased gradually. In addition, intratumoral mast cells expressed higher levels of the immunosuppressive molecule galectin-9, which effectively suppresses CD8+ T-cell antitumor immunity in vitro. Blocking galectin-9 on tumor-infiltrating mast cells reversed the immunosuppression of CD8+ T cells. In conclusion, our data elucidated novel protumorigenic and immunosuppressive roles of mast cells in TC. In addition, our results indicated that blocking mast cells may impede tumor progression and ameliorate the prognosis of TC patients.

Survival status of HIV/AIDS patients aged ≥ 50 years old receiving antiretroviral therapy in Shanxi, 2011-2019 / 中国热带医学

@#Abstract: Objective　To analyze the survival status of HIV/AIDS patients aged above 50 years receiving antiviral therapy (ART) in Shanxi Province from 2011 to 2019, and to provide evidence for further improvement of antiviral therapy. Methods　Basic information and follow-up information of HIV/AIDS patients aged above 50 years who first received HIV/AIDS antiviral therapy in Shanxi Province from 2011 to 2019 were collected. Excel database was established and SPSS23.0 software was used for analysis. Retrospective cohort study was conducted. Cox proportional risk regression model was used to analyze the factors influencing survival time. Results　A total of 1 183 subjects were included, of which 172 died, including 84(48.84%) from other causes, 74(43.02%) AIDS-related death and 14 (8.14%) from accidents, suicides and undetermined deaths. Setting AIDS-related deaths as an outcome event, life table analysis showed that the cumulative survival rates at 1, 3, 5, 7 and 9 years after receiving ART were 96.61%, 93.59%, 90.35%, 87.57% and 83.44%, respectively. Multivariate Cox proportional risk model analysis showed that the risk of death in patients aged 60-<70 years group and over 70 age group was 2.53 times (95%CI: 1.51-4.23) and 3.59 times (95%CI: 1.74-7.40) for patients aged the 50-<60 group , respectively. The risk of death in patients with baseline CD4+T lymphocyte (CD4) counts of ≥200/mm3, 50-<200 /mm3 was 0.22 times (95%CI: 0.12-0.41) and 0.37 times (95%CI: 0.21-0.67) for patients with CD4+T lymphocyte counts of <50/mm3. The risk of death in patients with opportunistic infections at baseline was 1.99 times (95%CI: 1.16-3.39) for patients without baseline opportunistic infections. Conclusions　The survival rate of HIV/AIDS patients aged above 50 who received antiviral therapy (ART) in Shanxi Province from 2011 to 2019 was relatively high. To further improve the quality of antiviral treatment in our province, the strategy of "early detection and early treatment" should be continued and improved in the future, and information collection of specific causes of non-AIDS-related deaths among this population should be further strengthened.

Influencing factors for the quality of bowel preparation with polyethylene glycol electrolyte powder combined with diet control before colonoscopy in children / 中国当代儿科杂志

OBJECTIVES@#To investigate the influencing factors for the quality of bowel preparation before colonoscopy in children and the association of the interval from the last administration of laxative to the start of colonoscopy (shortly referred to as waiting time) with the quality of bowel preparation.@*METHODS@#A retrospective analysis was performed for the children who were admitted to the Department of Gastroenterology, Children's Hospital of Nanjing Medical University, from January to November 2020, and received bowel preparation with polyethylene glycol electrolyte powder combined with diet control before colonoscopy. According to the score of Boston bowel preparation scale, they were divided into two groups: adequate bowel preparation group (n=337) and inadequate bowel preparation group (n=30). Related data were collected from the children in both groups, including general information, possible influencing factors for the quality of bowel preparation, adverse reactions associated with bowel preparation, duration of colonoscopy, and postoperative diagnosis. Univariate and multivariate analyses were used to explore the influencing factors for the quality of bowel preparation.@*RESULTS@#The univariate analysis showed that age, body weight, and waiting time were associated with inadequate bowel preparation (P<0.05). The multivariate analysis showed that older age (OR=2.155, 95%CI: 1.087-4.273, P=0.028) and longer waiting time (OR=1.559, 95% CI: 1.191-2.041, P=0.001) were independent risk factors for inadequate bowel preparation. The receiver operating characteristic (ROC) curve analysis showed that the cut-off value of waiting time was 5.5 hours in determining whether bowel preparation was adequate or not, with a sensitivity of 90.0%, a specificity of 50.7%, and an area under the ROC curve of 0.708. After grouping based on waiting time, it was found that the incidence rate of inadequate bowel preparation in the ≥5.5 hours group was significantly higher than that in the <5.5 hours group [14.0% (27/193) vs 1.7% (3/174), P<0.001].@*CONCLUSIONS@#For children who use polyethylene glycol electrolyte powder combined with diet control for bowel preparation, older age is an independent risk factor for inadequate bowel preparation before colonoscopy, which may be associated with an insufficient dose of polyethylene glycol in older children. Longer waiting time is also an independent risk factor for inadequate bowel preparation, and it is recommended that the waiting time should not exceed 5.5 hours.

The role of androgen in male sexual arousal / 中华男科学杂志

Sexual arousal is an important factor for the success of sexual behavior, and regulated by the central nervous system, its underlying mechanism is very complicated. Androgen is the most important endocrine hormone in men, which is deeply involved in the whole process of male sexual response, but how it regulates male sexual arousal has not been fully clarified and remains one of the hotspots in current andrological research. Therefore, this paper presents an overview of the advances in the studies of the related role and mechanism of androgen in male sexual arousal. In the central nervous system, androgen regulates the release of dopamine neurotransmitters by binding androgen receptors or metabolizing neurosteroids, thus activating the brain reward system. Besides, androgen regulates the neuronal plasticity and spinous process formation in the neural circuit of sexual arousal to ensure successful activation and conduction of the neural circuit. However, the specific regulating mechanism of sexual arousal remains to be further explored.

Treatment of superior mesenteric artery embolism by percutaneous mechanica thrombectomy with the Solitaire FR stent system / 中国实用外科杂志

OBJECTIVE: To evaluate the technical feasibility,safety,and clinical outcome of mechanical thrombectomy with Solitaire FR stent system for embolic occlusion of the superior mesenteric artery(SMA).METHODS: The clinical data of 6 patients with embolic occlusion of the SMA treated by mechanical thrombectomy with Solitaire FR stent system between January 2015 and June 2018 in Binzhou City People's Hospital were analyzed retrospectively.RESULTS: Superior mesenteric artery occlusion was initially diagnosed by computed tomography(CT)in all patients.A successful thrombus removal of superior mesenteric arterial by Solitaire FR stent system was observed in the 6 patients.Five patients had recovered well after operation and no complications such as artery dissection,perforation and hemorrhage or intestinal ischemia.One patient underwent bowel resection.CONCLUSION: The arterial mechanical thrombectomy with solitaire FR stent system are characterized with high rate of recanalization,fine security,minimal invasion and less complications in patients with acute superior mesentericvarterial embolism.

Necroptosis occurs in osteoblasts during tumor necrosis factor-α stimulation and caspase-8 inhibition

Necroptosis is a regulated cell death mechanism. However, it is unknown whether necroptosis is involved in the death of tumor necrosis factor-α (TNF-α)-treated osteoblasts. Therefore, we conducted the study with TNF-α, Nec-1 (a specific inhibitor of necroptosis), and Z-IETD-FMK (a specific inhibitor of apoptosis) to determine whether necroptosis plays a role in the death of TNF-α-treated osteoblast cell line MC3T3-E1. Cell viability, cell death, and lactate dehydrogenase (LDH) release were assayed to evaluate cytotoxicity. Specific marker proteins receptor interacting protein kinase (RIPK3) and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) for necroptosis, and cleaved caspase 3 for apoptosis were detected by western blot, and mRNA was measured by quantitative real-time polymerase chain reaction (qRT-PCR). We found that TNF-α inhibited cell proliferation in a dose- and time-dependent manner. Nec-1 plus Z-IETD-FMK restored cell viability and significantly decreased LDH release. In addition, TNF-α alone increased the cell population of AV+PI−, while Z-IETD-FMK caused a shift in the cell population from AV+PI− to AV+PI+. Furthermore, TNF-α significantly increased protein cleaved caspase 3. TNF-α plus Z-IETD-FMK significantly increased the proteins RIPK3 and MLKL phosphorylation in MC3T3-E1 cells, while the changes in mRNA levels of RIPK3, MLKL, and caspase 3 were not consistent with the changes in the corresponding protein expression levels. In conclusion, TNF-α induced preferentially apoptosis in osteoblast cell line and necroptosis played a decisive role when TNF-α-induced death was inhibited by the inhibitor of apoptosis. Combined treatment with Nec-1 and Z-IETD-FMK protected mouse osteoblasts from death induced by TNF-α.

Advances in tumor microenvironment and related targeted drugs / 药学学报

Tumor microenvironment (TME) is the internal environment of tumor. As a functional unit, the microenvironment determines the occurrence and development of tumors. Hypoxia, inflammation and immunosuppression are three major characteristics of TME. Hypoxia signals involve in multiple immunosuppressive pathway. There is communication among the gut microbiota, chronic inflammation and immunity. The three characteristics are associated with each other to form a complex network affecting the metastasis of the tumor. TME also influences the development of immunotherapy and efficacy of drugs in solid tumor. Therefore, it is important to identify the valuable biomarkers to predict disease progression, to elucidate the mechanistic networks in the microenvironment, to develop microenvironment targeting drugs and effective drug combination strategies to improve the drug efficacy. All of these have a profound clinical value.

Distinct Rab7-related Endosomal-Autophagic-Lysosomal Dysregulation Observed in Cortex and Hippocampus in APPswe/PSEN1dE9 Mouse Model of Alzheimer's Disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Amyloid-β deposition and accumulation of autophagic vacuoles are pathologic features of Alzheimer's disease (AD). Dysregulation of the endosomal-autophagic-lysosomal (EAL) pathway, which impairs amyloid precursor protein processing, is one of the earliest changes in AD. However, the precise role of EAL pathway in neurodegeneration remains unclear. This study aimed to investigate the role of EAL pathway in AD and further study the mechanism of EAL dysfunction.</p><p><b>METHODS</b>We used 3-, 7-, and 12-month-old APPswe/PSEN1dE9 (APP/PS1) mice to model different stages of AD with age- and gender-matched wild-type littermates as controls (4-7 mice per group) and detected the changes of EAL markers, endosomal organizers Rab5 and Rab7, autophagosome marker LC3B, and lysosomal proteins Lamp1/2 in cortex and hippocampus by immunohistochemistry and Western blotting analysis. To further explore the mechanism of EAL dysregulation in AD, components of the class III phosphatidylinositol 3-kinase (PI3KC3) complex, activators of Rab7 (Beclin1 and UVRAG), and the negative regulator of Rab7 (Rubicon) were also measured in this two brain regions.</p><p><b>RESULTS</b>In 7-month-old APP/PS1 brain that amyloid beta initiated to accumulate intracellularly, EAL pathway, and related PI3KC3 members, UVRAG and Beclin1 were upregulated both in cortex and hippocampus (all P < 0.05). By the age of 12 months old, when abundant amyloid plaques formed, EAL markers, UVRAG, and Beclin1 were also upregulated in the cortex (all P < 0.05). However, Rab7 was decreased significantly (P = 0.0447), accompanied by a reduction of its activating PI3KC complex component Beclin1 (P = 0.0215) and enhancement of its inhibiting component Rubicon (P = 0.0055) in the hippocampus.</p><p><b>CONCLUSIONS</b>Our study implies that EAL pathway, represented as Rab7 and its PI3KC3 regulators' expressions, showed temporal and spatial variation in brains at different stages of AD. It provides new insights into the role of EAL pathway in pathogenesis and indicates potential therapeutic targets in neurodegenerative diseases.</p>

A study on the effect of different family tobacco control patterns on cotinine level and acute respiratory infections among infants / 预防医学

Objective To explore the effect of different family tobacco control patterns on cotinine level and acute respiratory infections among infants.Methods A total of 300 infants were included,and were divided into 3 groups based on the tobacco control patterns:strictly tobacco -controlled group (97 cases),partly tobacco -controlled group (88 cases)and tobacco -uncontrolled group (1 1 5 cases).Urinary cotinine was measured in all participants.All participants were prospectively followed -up for 1 year,and the incidence of acute respiratory infections was recorded during the follow-up.Results The cotinine level of strictly tobacco -controlled group [0.45 ±0.21 (μg/L)]was significantly lower than the other two groups [1 .01 ±0.49(μg/L),1 .1 6 ±0.48(μg/L),P <0.05],and no significant differences were detected between the partly tobacco -controlled group and tobacco -uncontrolled group.The incidence of lower respiratory tract infection,not the upper respiratory tract infections,was significant different among the 3 groups (strictly tobacco -controlled group:1 8.75%;partly tobacco -controlled group:32.1 8%;tobacco -uncontrolled group:37.72%)(P <0.05).The number of upper respiratory tract infections and lower respiratory tract infections was significantly different among the three groups.The difference was significant between strictly tobacco -controlled group and partly tobacco -controlled group and between strictly tobacco -controlled group and tobacco -uncontrolled group (P <0.05).Conclusion Strict tobacco control could reduce the prevalence of passive smoking and the incidence of respiratory infections among infants.

Determination of ASC-J9 in rat blood by HPLC-MS/MS / 中国药学杂志

OBJECTIVE: To establish an HPLC-MS/MS method for determination of ASC-J9 in rat blood. METHODS: After liquid-liquid extraction, ASC-J9 was separated on a Symmetry Cl8 colume, with mobile phase of acetonitrile-water containing 0.1% formic acid and 10 mmol·L-1 ammonium for mate. The flow rate was 1.0 mL·min-1, mass shunt was 0.4 mL·min-1, and column temperature was main tained at 35℃. Quantification was performed in positive ion multiple-reaction-monitoring (MRM) mode. RESULTS: The calibration curve of ASC-J9 had good linearity in the concentration range of 3.54-1180 ng·mL-1. The extraction recovery rate was within 83.19% to 87.27%, and the intra-day and inter-day RSDs were both less than 8.89%. CONCLUSION: This method is specific, sensitive and suitable for determination of ASC-J9 in rat blood.

Association of interleukin-10 gene-592C/A polymorphism with concentration of interleukin-10 in children with bronchial asthma / 中华实用儿科临床杂志

Objective To explore the polymorphism of-592C/A of IL-10 gene promoter region in children with bronchial asthma and its relationship with serum concentration of IL-10.Methods Ninety-two children with bronchial asthma and 92 healthy children were selected for study,polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used for the analysis of-592C/A of IL-10 promoter region polymorphism.The frequencies of genotypes of IL-10 gene-592 locus (CC,CA and AA) and alleles were accounted respectively,and x2 test was used to analyze the difference between the groups.Enzyme-linked immunosorbent assay (ELISA) was used to measure the concentration of the serum IL-10,and F test and q test were used for statistical analysis.Results Compared with the healthy control group,there were significant differences in-592C/A polymorphism of IL-10 gene in asthma group.The frequencies of AA genotypes (56.5％) and A allele (73.9％) in asthma group were higher than those (34.8％,58.7％)in the control group,there were significant differences(x2 =9.32,P ＜ 0.01 ;x2 =8.87,P ＜ 0.005,respectively).The individuals with AA genotype and A allele were 3.25 (95 ％ CI:1.28-8.28,P ＜ 0.05) and 1.99 (95 ％ CI:1.28-3.08,P ＜0.01) times susceptible to asthma compared with CC genotype and C allele.The serum concentration of IL-10 in asthma group was significantly lower than that in healthy control group,whether in attacking-stage or remission-stage,and there were significant differences (all P ＜ 0.01).The individuals with AA genotypes had lower serum IL-10 concentration than those with CC genotypes (P ＜ 0.05) . Conclusions The IL-10 gene-592C/A polymorphism is different significantly between children with bronchial asthma and healthy ones,and this polymorphism influences the concentration of IL-10.The individuals with AA genotype have relatively lower IL-10 concentration,and A allele may be one of genetic susceptibility factor of bronchial asthma in children.

Host glial cell canceration induced by glioma stem cells in GFP/RFP dual fluorescence orthotopic glioma models in nude mice / 中华肿瘤杂志

<p><b>OBJECTIVE</b>During the process of tissue remodeling in human tumor transplantation models, the roles of the inoculated tumor cells and host tissue in tumor progression is still largely unknown. The aim of this study was to investigate the relationships and interactions between these two sides using GFP-RFP double fluorescence tracing technique.</p><p><b>METHODS</b>Red fluorescence protein (RFP) gene was stably transfected into glioma stem cell line SU3, then SU3-RFP cells were transplanted into the brain of athymic nude mice with green fluorescence protein (GFP) expression. After the intracerebral tumors were formed, the relationship and interaction between GFP cells and RFP cells were analyzed. Highly proliferative GFP cells were screened out, and monocloned with micro-pipetting. DNA content assay, chromosome banding and carcinogenicity test of the GFP cells were performed to observe the GFP cells' cancerous phenotype in nude mice.</p><p><b>RESULTS</b>In the transplantable tumor tissue, besides a great quantity of RFP cells, there were still a proportion of GFP cells and GFP/RFP fusion cells. The proportion of RFP cells, GFP cells and GFP/RFP cells were (88.99 ± 1.46)%, (5.59 ± 1.00)%, and (4.11 ± 1.020)%, respectively. Two monoclonal host GFP cells (H1 and H9) were cloned, which demonstrated the properties of immortality, loss of contact inhibition, and ultra-tetraploid when cultured in vitro. Both H1 and H9 cells expressed CNP, a specific marker of oligodendrocytes. The GFP cells also demonstrated 100% tumorigenic rate and high invasive properties in vivo.</p><p><b>CONCLUSIONS</b>In this glioma transplantation model, the transplanted tumor tissues contained not only transplanted glioma stem cells but also cancerous host GFP cells. Our findings offer important clues to further research on the relationships among different members in the tumor microenvironment.</p>

Relationship between spinal function and the severity of spinal cord injury by needle puncture / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the changes in spinal cord pathophysiology, motor function and electrophysiology after spinal cord injuries induced by punctures with different needles, and explore a new means for studying spinal neurotoxicity of local anesthetics.</p><p><b>METHODS</b>A total of 144 SD rats were randomly allocated into the sham-operated group (n=36) and 3 spinal cord injury groups (n=36) with the L4-5 segment of the dura mater of the spinal cord punctured using 29G, 25G, and 21G needles. The BBB scores before surgery were recorded, and at 8 h, 24 h, 72 h, 1 week, and 2 weeks after the surgery, the motor evoked potential (MEP), spinal cord pathology and the BBB scores were examined.</p><p><b>RESULTS</b>In the control group, the rats showed normal BBB score, spinal function and microstructure. Spinal cord puncture with 29G needle did not cause obvious pathologies of the spinal cord, whereas puncture with 21G needle resulted in marked changes in the motor function, electrophysiology and histology of the spinal cord, which showed significant improvements at 2 weeks postoperatively.</p><p><b>CONCLUSION</b>Puncture with a 29G needle causes less injuries and minimal functional changes of the spinal cord, which can serve as a new means for studying spinal neurotoxicity of local anesthetics.</p>

Incubation and application of transgenic green fluorescent nude mice in visualization studies on glioma tissue remodeling / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The primary reasons for local recurrence and therapeutic failure in the treatment of malignant gliomas are the invasion and interactions of tumor cells with surrounding normal brain cells. However, these tumor cells are hard to be visualized directly in histopathological preparations, or in experimental glioma models. Therefore, we developed an experimental human dual-color in vivo glioma model, which made tracking solitary invasive glioma cells possible, for the purpose of visualizing the interactions between red fluorescence labeled human glioma cells and host brain cells. This may offer references for further studying the roles of tumor microenvironment during glioma tissue remodeling.</p><p><b>METHODS</b>Transgenic female C57BL/6 mice expressing enhanced green fluorescent protein (EGFP) were crossed with male Balb/c nude mice. Then sib mating was allowed to occur continuously in order to establish an inbred nude mice strain with 50% of their offspring that are EGFP positive. Human glioma cell lines U87-MG and SU3 were transfected with red fluorescent protein (RFP) gene, and a rat C6 glioma cell line was stained directly with CM-DiI, to establish three glioma cell lines emitting red fluorescence (SU3-RFP, U87-RFP, and C6-CM-DiI). Red fluorescence tumor cells were inoculated via intra-cerebral injection into caudate nucleus of the EGFP nude mice. Tumor-bearing mice were sacrificed when their clinical symptoms appeared, and the whole brain was harvested and snap frozen for further analysis. Confocal laser scanning microscopy was performed to monitor the mutual interactions between tumor cells and host brain cells.</p><p><b>RESULTS</b>Almost all the essential tissues of the established EGFP athymic Balb/c nude mice, except hair and erythrocytes, fluoresced green under excitation using a blue light-emitting flashlight with a central peak of 470 nm, approximately 50% of the offsprings were nu/nu EGFP+. SU3-RFP, U87-RFP, and C6-CM-DiI almost 100% expressed red fluorescence under the fluorescence microscope. Under fluorescence microscopic view, RFP+ cells were observed growing wherever they arrived at, locating in the brain parenchyma, ventricles, and para-vascular region. The interactions between the transplanted tumor cells and host adjacent cells could be classified into three types: (1) interweaving; (2) mergence; and (3) fusion. Interweaving was observed in the early stage of tumor remodeling, in which both transplantable tumor cells and host cells were observed scattered in the tumor invading and spreading area without organic connections. Mergence was defined as mutual interactions between tumor cells and host stroma during tumorigenesis. Direct cell fusion between transplantable tumor cells and host cells could be observed occasionally.</p><p><b>CONCLUSIONS</b>This study showed that self-established EGFP athymic nude mice offered the possibility of visualizing tumorigenesis of human xenograft tumor, and the dual-color xenograft glioma model was of considerable utility in studying the process of tumor remodeling. Based on this platform, mutual interactions between glioma cells and host tissues could be observed directly to further elucidate the development of tumor microenvironment.</p>

Expression of miR-125b in the new, highly invasive glioma stem cell and progenitor cell line SU3 / 癌症

MicroRNA (miR)-125b has been shown to play a potential role in the development of glioma stem cells. However, the relationship between miRNA and glioma stem cells is still elusive. This study was designed to elucidate this potential relationship. We established a highly invasive glioma stem cell and progenitor (GSCP) cell line SU3. SU3 cell suspensions were injected into nude mice brains in situ, and the invasiveness of graft tumors was analyzed using hematoxylin and eosin staining as well as immunohistochemistry. Real-time polymerase chain reaction (PCR) was used to measure the expression levels of miR-125b in SU3 and other cells. In vitro, SU3 cells expressed CD133 and nestin as well as differentiation markers glial fibrillary acidic protein (GFAP) and β-tubulin III, which were consistent with the characteristics of glioma stem cells. Scratch assays indicated that the migration ability of SU3 cells was stronger than that of U251 stem cells (U251s). In vivo, SU3 cells invaded into each part of the mouse brain from the caudate nucleus in a diffuse pattern and highly expressed invasive and proliferative cell markers matrix metalloprotease 2 (MMP2), MMP9, and Ki-67. Real-time PCR results revealed that the levels of miR-125b and MMP9 were significantly higher in SU3 and SU2, also a highly invasive GSCP cell line we established before, than in U251s. High expression of miR-125b both in newly established GSCPs, SU3, and long-term cultured GSCPs, SU2 suggests that miR-125b exhibits oncogene-like behavior. This behavior should be considered in further studies of miR-125b in cancer stem cells. Furthermore, MMP9, which plays a role in cancer stem cell invasion, may be a target gene of miR-125b.

Association of leptin level and leptin receptor gene polymorphisms with susceptibility to severe pre-eclampsia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the association of serum leptin concentrations and polymorphisms of G1019A and A223G of leptin receptor gene (LEPR) with severe pre-eclampsia. MEHTODS: A case-control study was carried out in 207 patients with severe pre-eclampsia (SPE group) and 252 healthy pregnant women (control group) during the third trimester of pregnancy. The serum leptin was determined by enzyme-linked immunosorbent assay. The polymorphisms of LEPR gene G1019A and A223G were detected by polymerase chain reaction restriction-fragment length polymorphism (PCR-RFLP) analysis. Miettinen's test was used to estimate the odds ratios (OR) and 95% confidence intervals (CI).</p><p><b>RESULTS</b>(1) In severe pre-eclampsia group, serum leptin levels and rate of premature infant birth were significantly higher than that in normal pregnant women, and birth weight was lower than that in controls (P<0.01). (2) The frequencies of GA genotype and G allele for LEPR gene G1019A in SPE group (33.8% and 20.3%) were markedly higher than that in controls (19.8% and 15.1%) (P<0.01), and the carriers of GA genotype and G allele were more frequent in SPE group than in control group, resulting in an OR 2.04 (95%CI: 0.77-5.42) and 1.43 (95%CI: 1.02-2.01) to develop severe pre-eclampsia, compared with carriers of AA genotype and A allele. (3) AG genotype and A allele frequencies of LEPR gene A223G in SPE group (19.3% and 12.6%) were significantly lower than that in controls (34.5% and 19.2%) (P<0.01), resulting in an OR of 0.46 (95%CI: 0.30-0.71) and 0.60 (95%CI: 0.42-0.87) to develop severe pre-eclampsia, compared with subjects with GG genotype and G allele. (4) The "1019AA+223AG" genotype frequency was significantly lower in SPE group (6.8%) than in controls (24.6%) (P<0.01), resulting in an OR of 0.22 (95%CI: 0.12-0.39) to develop severe pr-eclampsia, while the "1019AA+223AG" was significantly higher in SPE group (22.2%) than in controls (11.9%) (P<0.05), resulting in an OR of 2.10 (95%CI: 0.78-3.45) to develop severe pre-eclampsia. (5) No significant differences were found in SBP, DBP, BMI and serum leptin levels in subjects with different genotypes in the two groups (P>0.05).</p><p><b>CONCLUSION</b>Elevated serum leptin level and LEPR gene G1019A and A223G polymorphisms might play a role in severe pre-eclampsia, while the level of serum leptin was not associated with genotypes of LEPR gene G1019A and A223G polymorphisms. The genotypes GA and "1019AA+223AG"of G1019A may be genetic susceptibility factors to severe pre-eclampsia.</p>

Preliminary studies on cell derivation of neovascularization in human glioma and its functional evaluation / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The finding of vasculogenic mimicry (VM) in many solid tumors indicates that tumor cells themselves could participate in the construction of tumor vessels. However the origin of these cells is still not fully elucidated, and whether these vessels have the ability of blood-supply is still unclear. Preliminary studies were performed to investigate whether part of tumor neovascularity is derived from tumor stem cells (TSCs) and whether TSCs-derived vessels are functional.</p><p><b>METHODS</b>Transplanted glioma tissues obtained from subcutaneous and orthotopic transplantation nude mouse models were processed into paraffin sections. In order to identify the cell origin and types of tumor vessels, sections were stained with CD31, CD34, CD133, GFAP, Ki67 and HLA, respectively. CD34-PAS staining was performed as well. A part of tumor-bearing mice were perfused with activated carbon through the systemic circulation and the distribution of activated carbon was observed.</p><p><b>RESULTS</b>CD34-PAS staining showed that endothelium-dependent vessels (CD34(+), PAS(+)), VM vessels (CD34(-), PAS(+)), and the MVs (CD34(+), PAS(-)) could be seen in the transplantated tumors. Activated carbon particles were observed in all three types of vessels. CD31(+) cells adherent to the luminal surface of vessel wall. CD34(+) cells distributed along the vessels as well, but morphologically were more like a transition type between tumor cells and endothelial cells. Human specific Ki67 and HLA positive cells could be seen in the tumor vessels indicating that these vessels were derived from human tumor cells. Moreover, cells of tumor vessels were proved to be constructed by human tumor cells mainly and fusion cells of host cells and tumor cells under confocal microscope.</p><p><b>CONCLUSIONS</b>Three types of blood supply sources including endothelium-dependent vessels, vasculogenic mimicry (VMs) and mosaic vessels (MVs) exist in transplantation tumors of human glioma. Glioma stem and progenitor cells (GSCPs) have the potential to differentiate and transdifferentiate into VMs and MVs.</p>

Isolation and multipotent differentiation of human decidua basalis-derived mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the biological features of human decidua basalis-derived mesenchymal stem cells (PDB-MSCs) in vitro and identify their capacity of multilineage differentiation.</p><p><b>METHODS</b>PDB-MSCs were harvested from the decidua basalis of term placental by enzymatic digestion and density gradient centrifugation, and the growth characteristics and morphological changes of the MSCs were observed by inverted microscope. The proliferative ability of the cells was assessed by Cell Counting Kit-8. The cell cycle and expressions of the surface markers (CD29, CD44, CD73, CD90, CD34, CD45, and CD14) of the MSCs were identified by flow cytometry. Multilineage differentiation capacity of the cells was tested by inducing their differentiation toward osteoblasts, adipocytes and chondroblasts in vitro.</p><p><b>RESULTS</b>MSCs isolated from human decidua basalis of term placental exhibited a morphology similar to that of bone marrow-derived MSCs, and grew into colonies in in vitro culture, where the cells proliferated rapidly after passage with a cell doubling time of 2.21∓0.21 days. More than 70% of the cells stayed in the resting stage (G(0)/G(1)) and showed positivity for CD29, CD44, CD73 and CD90, but not for CD14, CD34 or CD45. After induction, the cells showed positive results of alizarin red staining, oil red O staining and Alcian blue staining.</p><p><b>CONCLUSION</b>Human decidua basalis contains a rich source of MSCs, which can be easily isolated and cultured without affecting their capacity of multilineage differentiation. The PDB-MSCs may have the potential as a novel source of stem cells.</p>

Construction of a mouse pDsRed2-N1-SDF-1alpha eukaryotic expression vector and its expression in mouse bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector pDsRed2-N1-SDF-1alpha and observe its expression in the mouse bone marrow mesenchymal stem cells.</p><p><b>METHOD</b>SDF-1alpha gene sequence with XhoI, EcoRI restriction enzyme cutting site was amplified from the total RNA of mouse smooth muscle cells by reverse transcription-polymerase chain reaction (RT-PCR) and inserted into the eukaryotic expression vector pDsRed2-N1 encoding red fluorescent protein gene, and the insertion was verified by endonuclease digestion and DNA sequencing. Mouse bone marrow mesenchymal stem cells identified with immunofluorescence assay for vimentin expression were transfected with the constructed plasmid pDsRed2-N1-SDF-1alpha, and the expression of sdf-1alpha was detected using immunofluorescence assay.</p><p><b>RESULTS</b>The DNA fragment amplified by PCR from the total RNA was identical to SDF-1alpha from the gene library, and an identical DNA fragment was also amplified from the recombinants. Sequence analysis confirmed the successful insertion of SDF-1alpha into the pDsRed2-N1 vector and the eukaryotic expression vector pDsRed2-N1-SDF-1alpha was successfully constructed. The cultured mouse bone marrow mesenchymal stem cells positive for vimentin protein showed SDF-1alpha expression 24 h after transfection with the recombinant vector.</p><p><b>CONCLUSION</b>The pDsRed2-N1-SDF-1alpha eukaryotic expression vector constructed is capable of expression of SDF-1alpha fusion protein in the mouse bone marrow mesenchymal stem cells.</p>

Clinical factors related to bone metastases from breast cancer / 中华核医学杂志

Objective To study the clinical and imaging features of patients with bone metastases from breast cancer and identify the factors related to the incidence of bone metastases. Methods Three hundred and thirty-four patients with breast cancer were recruited into this study. Whole-body 99Tcm-methylene disphosphonate (MDP) bone scan, clinical staging, pathological, immunohistochemical and serological test results were analyzed retrospectively. χ2 test was used for statistical analysis. Results The incidence rate of bone metastases for patients with and without lymph node metastases was 71% (152/214) and 22. 5% (27/120), respectively (χ2 =72.80, P =0.000). The incidence rate of bone metastases from infiltrated non-specified and specified breast cancer was 69% (203/294) and 41.7% (5/12), respectively (χ2 =3. 97, P=0.046). Alkaline phosphatase (ALP) was elevated in 28.5% (51/179) and 14.9%(11/74) of patients with and without bone metastases, respectively (χ2 = 5. 25, P = 0.022 ). Carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 15-3, CA125, CA19-9 increased in 68.7% ( 123/179) and 27.0% (20/74) of patients with and without bone metastases, respectively (χ2 = 37. 03, P =0. 000). Conclusions The incidence of bone metastases from breast cancer is correlated to pathological types of primary tumor and lymph node metastases. Bone metastases occurs more frequently in patients with infiltrated, non-specified, primary cancer and with lymph node metastases. Serum ALP, CEA, CA15-3,CA125, CA19-9 might be the tumor makers for early diagnosis of bone metastases from breast cancer.