Objective To evaluate the protective effectiveness of intranasal immunizations with recombinated-pneumococcal autolysin(Re-LytA), which protects mice against local and systemic Streptococ- cus pneumoniae(Sp) infection. Methods Testing group (group A): CpG as an adjuvant, the mice were intranasally immunized with purified Re-LytA, obtained by affinity chromatograph. The negative control group(group B) were intranasally immunized with sterile saline. And the positive control group (group C) were received 23-valent polysaccharide commercial vaccine through intramuscular injection. All the samples were collected 2 weeks post the last immunization. The levels of antibody was determined by ELISA. Then the mice were challenged intraperitoneally and intranasally with Sp, respectively. The infection and coloniza- tion was followed by monitoring colony-forming units of Sp in the blood, homogenized lung, and nasopharyn- geal lavage fluid 4 days post intranasal immunization. The mice were observed daily to note the livability of each group. Results The level of the LytA antibody (IgG, IgA, slgA) in group A were higher than that in group B and C (P < 0.05). Neither the LytA nor polysaccharide antibody could be detected in group B. Polysaccharide antibody could be detected in group C. After challenged intraperitoneally there was no signifi- cant difference in survival rates between group A and group C (P > 0.05), which was significant higher than that in group B (P <0.05). After challenged intranasally, compared with the group A, the geometric mean colony-forming units washed from the nasopharyngeal lavage fluid of the group B and group C were signifi- cantly higher (P <0.05). Conclusion lntranasal immunizations with Re-LytA can protect mice against lo- cal and systemic pneumococcal infection, and the protective immunity may be related to sIgA.

Objective　To investigate the protective effect of calcium antagonist Verapamil (VP) on kidney preservation in HCA solution. Methods　After kidneys were isolated from rabbits, they were perfused and stored in HCA solution or in HCA solution with VP pre-supplement at 4℃ for 24 h respectively. The contents of mitochondrial calcium in renal cells and ATP in renal tissues were measured in every group. Results　The contents of mitochondrial calcium was remarkably higher and ATP significantly lower in the kidneys in HCA solution at 4℃ for 24 h than those just after resection. But these could be inhibited in those storing in the HCA solution with VP pre-supplement. Conclusion　Calcium antagonist VP can protect kidney function during HCA solution preservation by inhibiting calcium intaking into mitochondrium.

Objective To observe the effects of acupoint catgut embedding therapy on body mass,lipid metabolism,serum leptin and mRNA and protein expressions of hypothalamic leptin receptor(LepR)-mediated Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway in rats with diet-induced obesity(DIO).Methods Forty male SD rats were randomly divided into 10 in normal group and 30 in modeling group.A high-fat diet was used to establish the DIO rat model.After successful modeling,the modeled rats were randomly divided into the model group,the acupoint catgut embedding group and the acupoint catgut embedding + AG490(JAK2/STAT3 pathway blocker)group,with 10 rats in each group.The acupoint catgut embedding group and the acupoint catgut embedding + AG490 group were embedded on day(s)1,8,15 and 22 after successful modeling,the acupoints were selected from the Zhongwan(RN12),Shuidao(ST28),Tianshu(ST25),Pishu(BL20),Weishu(BL21),Sanjiaoshu(BL22)with a total of 4 treatments,and the acupoint catgut embedding + AG490 group was injected intraperitoneally with 1 mg/kg of AG490 every day during the treatment period;the normal group and the model group were only grasped and fixed.Body mass was measured before and after treatment.Lipid metabolism indexes of triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),and serum leptin levels were measured after treatment,and the mRNA expressions of hypothalamus LepR,JAK2 and STAT3 were detected by real-time quantitative polymerase chain reaction(RT-PCR),and the protein expressions of hypothalamus LepR,JAK2 and STAT3 were detected by Western Blot.Results Before treatment,compared with the normal group,the body mass of the model group,the acupoint catgut embedding group,and the acupoint catgut embedding+AG490 group were all elevated(P＜0.01),and compared with the model group,there was no significant difference in the body mass between the acupoint catgut embedding group and the acupoint catgut embedding+AG490 group(P＞0.05).After treatment,compared with the normal group,body mass,leptin and TG,TC,LDL-C levels were increased,and mRNA and protein expression levels of LepR,JAK2,STAT3 were decreased in the model group(all P＜0.01);compared with the model group,body mass,leptin and TG,TC,LDL-C levels were decreased in the acupoint catgut embedding group,and mRNA and protein levels of LepR,JAK2,STAT3 were increased in the acupoint catgut embedding + AG490 group(all P＜0.01);compared with the acupoint catgut embedding + AG490 group,the body mass,leptin and TG,TC,LDL-C levels were decreased,and mRNA and protein levels of LepR,JAK2,STAT3 were increased in the acupoint catgut embedding group(P＜0.05 or P＜0.01).Conclusion Acupoint catgut embedding has a good effect on weight loss and lipid reduction in DIO rats,and its central mechanism may be related to the down-regulation of serum leptin level and activation of hypothalamic LepR-mediated JAK2/STAT3 pathway.

Background and Objective:Radiotherapy(RT)is a major nonsurgical modality in the comprehensive treatment for colorectal adenocarcinoma.The radioresistance of cancer stem cells(CSCs)is a key factor that influences therapeutic effectiveness.This study was to investigate the effects of specific chromosome structure and histone modification in CSCs in colorectal adenocarcinoma radioresistance.Methods:Samples were collected from resected human colorectaI adenocarcinomas.Subcutaneous colorectal cancer model was established in nude mice.Immunohistochemistry showed that xenografts generated from bulk colorectaI cancer cells resembled the original tumor specimen.Flow cytometry was performed to sort CSCs (CD133~+)and non-CSCs(CD133~-)from both resected samples of colorectal adenocarcinoma and xenograft before and after high single-dose radiation.The markers labeling heterochromatin(H3K9me3,HP1-α,and H3K4me1)and euchromatin(H3K4me3)in CD133~+ and CD133-nucleus were detected by immunoflurescence.Results:There was distinct difference in chromatin structure between colorectaI CSCs(CD133~+)and non-CSCs (CD133~-).The chromatin displayed compact patches in CD133~+ nucleus,but loosely latticed structure in CD133-nucleus:immunoflurescence verified that the compact patches existing in CSCs was generated from heterochromatin construction.In addition.the vacuole-like defect in heterochromatin regions of CSCs was observed within 24 h after exposure to 10 gray(Gy)single-dose RT.Interestingly.this phenomenon was repaired from 96 h.and recovered to dense plaque structure in heterochromatin regions of CSCs after 144 h.However,no significant difference in non-CSCs was observed after RT exception for a loose chromatin structure.Conclusions:CSCs play a role in radiosensitivity in colorectal cancer.The mechanism may be related to heterchromatin formation and histone methylation.

Immunophenotype is critical for diagnosing common B-cell acute lymphoblastic leukemia (common ALL) and detecting minimal residual disease. We developed a protocol to explore the immunophenotypic profiles of common ALL based on the expression levels of the antigens associated with B lymphoid development, including IL-7Rα (CD127), cytoplasmic CD79a (cCD79a), CD19, VpreB (CD179a), and sIgM, which are successive and essential for progression of B cells along their developmental pathway. Analysis of the immunophenotypes of 48 common ALL cases showed that the immunophenotypic patterns were highly heterogeneous, with the leukemic cell population differing from case to case. Through the comprehensive analysis of immunophenotypic patterns, the profiles of patient-specific composite leukemia cell populations could provide detailed information helpful for the diagnosis, therapeutic monitoring, and individualized therapies for common ALL.
Adult ; Antigens, CD19 ; metabolism ; B-Lymphocytes ; immunology ; metabolism ; CD79 Antigens ; metabolism ; Female ; Humans ; Immunoglobulin Light Chains, Surrogate ; metabolism ; Immunophenotyping ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; pathology ; Receptors, Interleukin-7 ; metabolism

BACKGROUND AND OBJECTIVERadiotherapy (RT) is a major non-surgical modality in the comprehensive treatment for colorectal adenocarcinoma. The radioresistance of cancer stem cells (CSCs) is a key factor that influences therapeutic effectiveness. This study was to investigate the effects of specific chromosome structure and histone modification in CSCs in colorectal adenocarcinoma radioresistance.

METHODSSamples were collected from resected human colorectal adenocarcinomas. Subcutaneous colorectal cancer model was established in nude mice. Immunohistochemistry showed that xenografts generated from bulk colorectal cancer cells resembled the original tumor specimen. Flow cytometry was performed to sort CSCs (CD133+) and non-CSCs (CD133-) from both resected samples of colorectal adenocarcinoma and xenograft before and after high single-dose radiation. The markers labeling heterochromatin (H3K9me3, HP1-alpha and H3K4me1) and euchromatin (H3K4me3) in CD133+ and CD133- nucleus were detected by immunofluorescence.

RESULTSThere was distinct difference in chromatin structure between colorectal CSCs (CD133+) and non-CSCs (CD133-). The chromatin displayed compact patches in CD133+ nucleus, but loosely latticed structure in CD133- nucleus; immunofluorescence verified that the compact patches existing in CSCs was generated from heterochromatin construction. In addition, the vacuole-like defect in heterochromatin regions of CSCs was observed within 24 h after exposure to 10 gray (Gy) single-dose RT. Interestingly, this phenomenon was repaired from 96 h, and recovered to dense plaque structure in heterochromatin regions of CSCs after 144 h. However, no significant difference in non-CSCs was observed after RT exception for a loose chromatin structure.

CONCLUSIONSCSCs play a role in radiosensitivity in colorectal cancer. The mechanism may be related to heterochromatin formation and histone methylation.

AC133 Antigen ; Adenocarcinoma ; pathology ; radiotherapy ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Nucleus ; genetics ; pathology ; Colorectal Neoplasms ; pathology ; radiotherapy ; Female ; Glycoproteins ; metabolism ; Heterochromatin ; metabolism ; Histones ; metabolism ; Humans ; Male ; Methylation ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Neoplastic Stem Cells ; pathology ; radiation effects ; Particle Accelerators ; Peptides ; metabolism ; Radiation Dosage ; Radiation Tolerance

AIMTo explore the effect of Triptolide on airway remodeling and the expression of Phosphoinositide 3-Kinases in asthmatic rats.

METHODS40 rats were randomly divided into 5 groups (n = 8): (1) Control group; (2) Asthmatic 4 weeks group; (3) Asthmatic 6 weeks group; (4) Therapeutic 4 weeks group; (5) Therapeutic 6 weeks group. The airway resistance and eosinophilic inflammation of airway wall were observed. The airway wall thickness (WA/Pi), the bronchial smooth muscle thickness (smooth muscle area/Pi) and the number of bronchial smooth muscle nucleus (N/Pi) were measured by image analysis system. The expression of PI3K protein and mRNA were determined by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS(1) The expression of PI3K p85alpha protein and mRNA in asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01 P < 0.05). (2) The WA/Pi, the smooth muscle area/Pi and the N/Pi of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01). (3) The airway resistance of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than the control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01, P < 0.05).

CONCLUSIONThe proliferation of airway smooth muscle is a remarkable character of airway remodeling in asthma. The PI3K signal pathway may be involved in the process. Triptolide may reduce AHR and decrease the proliferation of ASMCs by inhibiting the expression of PI3K. It may have potential therapeutic effects in the asthmatic airway remodeling.

Airway Remodeling ; Animals ; Asthma ; metabolism ; physiopathology ; Diterpenes ; pharmacology ; Epoxy Compounds ; pharmacology ; Male ; Phenanthrenes ; pharmacology ; Phosphatidylinositol 3-Kinase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Child ; Humans ; Liver Diseases ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Portal Vein ; pathology

Objective To investigate the effects of rhynchophylline on methamphetamine-dependent SH-SY5Y cells model and microRNA-375-3p(miR-375-3p)/embryonic lethal abnormal vision drosophila-like 4(Elavl4)expression.Methods A methamphetamine-dependent cell model by maximum safe dose induction was established.The cells were divided into normal group(complete culture medium),control group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 48 h),model group(complete culture medium+100 μmnol·L-1 methamphetamine incubated for 48 h)and experimental group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 15 min,then 100 μmol·L-1 methamphetamine incubated for 48 h).The cyclic adenosine monophosphate(cAMP)and 5-hydroxytryptamine(5-HT)expression were detected by enzyme-linked immunosorbent assay(ELISA),miR-375-3p expression was detected by quantitative real time polymerase chain reaction(qRT-PCR),and Elavl4 expression was detected by Western blot.Results The cAMP expression levels of normal group,control group,model group and experimental group were(6.33±0.93),(6.57±1.12),(10.89±1.03)and(7.81±1.32)pmol·mg-1;5-HT were(682.46±17.32),(690.31±15.09),(510.11±27.67)and(649.99±21.42)pg·mL-1;miR-375-3p expression were 1.00±0.13,1.13±0.24,3.48±0.18 and 1.58±0.19;Elavl4 expression were 1.00±0.05,0.89±0.10,0.50±0.09 and 0.90±0.11,respectively.The differences between above indicators in model group and normal group were statistically significant(P＜0.01,P＜0.05);the differences between above indicators in experimental group and model group were statistically significant(P＜0.01,P＜0.05).Conclusion This study preliminarily established a methamphetamine-dependent cell model,and also found that rhynchophylline may regulate miR-375-3p/Elavl4 expression to antagonize methamphetamine addiction.

Objective To explore the effect of nest-type care combines with non-nutritive sucking on the growth and development of low birth weight infant.Methods According to whether supplemented the" nest"type of care and non-nutritive sucking,totals of 160 cases of low birth weight infants were divided into the observation group( n =80) received the nest-type care combines with non-nutritive sucking,and the control group( n =80) received traditional nursing.Then,the growth,the occurrence of feeding intolerance,the ease of breathing pauses of infant were observed.Results The average time of infant in the observation group returned to birth weight was (4.± 0.6) days four days significantly shorter than (7.2 ± 2.3) days in the control group (t =11.29,P＜0.01 ).The seventh and fourteenth days after admitted to hospital,the weight of infant in observation group were respectively ( 106.0 ± 12.2),(299.7± 3.4) g,which increased significantly faster than that in the control group respectively ( 31.0 ± 16.5 ),(235.3 ± 0.5 ) g,and the difference was statistically significant (P＜0.01 ).Sleep time of seventh and fourteenth days after admitted were respectively (20.6 ±2.4) hours and (20.4 ±1.8 ) hours,which was better than that in the control group ( 19.2 ± 2.2 ) hours and ( 19.1 ± 1.2) hours,and the difference was statistically significant ( t =3.85,5.37,respectively; P ＜ 0.01 ).The occurrence of feeding intolerance such as gastric retention,abdominal distension,vomiting,and apnes cases in the observation group significantly lower than that in the control group ( x2 =14.41,P ＜ 0.01 ).Conclusions Nesttype care combines with non-nutritive sucking make low birth weight infant feelling warmth and safety,It can improve the tolerance of enteral feeding,accelerate physical growth,and promote the low birth weight infant to grow and development.