Objective: To investigate the effects of Tongsaimai Prescription (TSMP) and TSMP without Achyranthis Bidentatae Radix (ABR) on ischemic brain injuries in rats. Methods: The male SD rats were divided into Sham, model, Nimodipine (32.4 mg/kg), TSMP (crude drug 31.74 g/kg), and TSMP without ABR (crude drug 27.77 g/kg) groups. Except the Sham and model groups, the rats were ig administered once daily, for 3 d. After the treatment, except the Sham group, the cerebral ischemic stroke model was established. After 24 h, the blood and brain tissue were taken, the brain tissue was stained with TTC, and the cerebral infarction ratio was measured. The pathological changes were observed, and the contents of HMGB1, TNF-α, and IL-1β in serum were detected. The expression of NF-κB in ischemia tissue was observed by immunohistochemisty method. Results: The expression of HMGB1, TNF-α, and IL-1β increased obviously (P < 0.05). TSMP and TSMP without ABR could decrease the contents of HMGB1, TNF-α, and IL-1β, inhibit the expression of NF-κB (P < 0.05), and the effect of TSMP without ABR on decreasing IL-1β was stronger. Conclusion: Both TSMP and TSMP without ABR could inhibit the activation of HMGB1-related NF-κB signal pathway with the function of anti-imflammation, alleviated encephaledema, and protecting nerve cells. TSMP without ABR has the similar effect on the ischemic brain injuries and the better effect on inhibiting the secretion of inflammatory factor IL-1β.

Abstract: Objective To explore the prevalence and bacterial strains of Francisella tularensis (F. tularensis) in wild rodents in Changbai Mountain area of China, and to further understand the epidemiological characteristics of F. tularensis infections in this area. Methods Wild rodents were captured from forest and forest-edge farmland from Kuandian County and Jianshi Forest District in the Changbai Mountain area, 2012-2014. DNA was extracted from the spleen tissues of the rodents, and the fopA gene of F. tularensis in wild rodents was detected using nested PCR. The infection rates were calculated for different areas and rat species. The bacterial subspecies of positive samples were identified using type-specific primers (C1/C4), and sequencing and comparative analysis were performed. Results A total of 133 wild rodents belonging to 6 rat species were captured. Among them, eight samples from three rat species (Apodemus agrarius, Apodemus peninsulae, Tscherskia triton ) were detected positive, with the overall positive rate of 6.01%. The positive rates of F. tularensis of Ji'an and Kuandian were 7.46% and 4.54%, respectively, and there was no difference in positive rates for different regions (χ2=0.117, P=0.732) and different rat species (χ2=0.641, P=0.986). The subspecies analysis showed that the detected 8 trains of F.tularensisall belonged to F.tularensis type B (F.subspecies subsp. holarctica). Genetic evolution analysis was performed on the fopA gene sequences of three positive samples (JA56, JA33, and JA38), which clustered together with Russia strains(CP009694.1, CP044004.1) and China strains (HM371344.1, HM371343.1) F.tularensis type B, with sequence similarities ranging from 99.21% to 99.47%. Conclusions Infection of F.tularensis subsp. holarctica existed in wild rodents in Changbai Mountain area of China, which suggests the existence of F.tularensis infection risks in this area.

OBJECTIVE: To study the effect and action mechanism of the large plant Rhodiola Tablet on myocardial ischemia of rats. METHODS: Rat myocardial ischemia models were induced by coronary artery ligation. The levels of AST, ALT, CK and CK-MB in serum were observed, at the same time the expressions of B-raf, Ras and p-ERK1/2 in myocardial tissue were determined. RESULTS: The levels of AST, ALT, CK and CK-MB in rat serum, as well as the B-raf, Ras and p-ERK1/2 expression in rats myocardium in the large plant Rhodiola group were significantly lower than that in the model group. CONCLUSION: Large plant Rhodiola has a certain antagonism on myocardial ischemia injury in rats caused by coronary artery ligation. The mechanism of action may be related to the decreases in serum AST, ALT, CK and CK-MB, as well as myocardial high expression of the B-raf, Ras and p-ERK1/2.

OBJECTIVETo investigate whether WASP/Verprolin homologous protein 1 (WAVE1) plays a role in the pathogenesis of childhood acute lymphoblastic leukemia (ALL).

METHODSWAVE1 mRNA and protein expression in bone marrow mononuclear cells (BMMCs) was measured by RT-PCR and Western blotting respectively in 4 children with ALL relapse, 15 children with ALL in complete remission (CR) and 40 children with newly diagnosed ALL. Ten normal bone marrow samples were used as controls. Jurkat cells were treated with different concentrations of adriamycin (ADM). The cell proliferation was detected with MTT. The apoptosis rate was measured by flow cytometry. WAVE1 mRNA and protein expression of Jurkat cells treated with ADM was detected by RT-PCR and Western blotting respectively.

RESULTSWAVE1 was not expressed or weakly expressed in BMMCs from normal controls and patients with ALL in CR. Higher WAVE1 mRNA and protein expression was found in BMMCs from patients with newly diagnosed ALL and patients with relapse ALL when compared with the controls and the patients in CR (P<0.01). ADM significantly inhibited the proliferation of the Jurkat cells and the inhibitory effect was dose-and time-dependent (P<0.05). After ADM treatment for 24 hrs, the percentage of apoptosis cells increased significantly and WAVE1 mRNA and protein expression of Jurkat cells decreased significantly when compared with the untreated controls (P<0.05).

CONCLUSIONSThe WAVE1 expression increased in children with ALL. WAVE1 may be related to the development of ALL and may be severed as a marker for the evaluation of the severity of ALL in children.

Adolescent ; Antibiotics, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Cell Proliferation ; drug effects ; Child ; Child, Preschool ; Doxorubicin ; pharmacology ; Female ; Humans ; Infant ; Jurkat Cells ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; etiology ; metabolism ; RNA, Messenger ; analysis ; Wiskott-Aldrich Syndrome Protein Family ; analysis ; genetics ; physiology

OBJECTIVETo investigate if WAVE1 is involved in mult drug-resistance (MDR) of human leukemia cell line K562/A02.

METHODSThe level of WAVE1 in K562 and K562/A02 cells was assayed by Western blot and RT-PCR; K562 cells and K562/A02 cells were transient transfected with pEFBOS-WAVE1 reconstructed plasmid or specifically siRNA to WAVE1. 50% inhibition concentration (IC50) of doxorubicin on K562/A02 was determined by WST-8 assay. Hoechst33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. The mRNA level of mdrl was assayed by RT-PCR. The Bcl-2 protein was assayed by Western blot.

RESULTS1. The WAVE1 expression at mRNA and protein level in K562/A02 cells was increased by about 70% and 63% respectively as compared with that in K562 cells. 2. Overexpression of WAVE1 in K562 cells by transient transfection significantly increased the resistance to doxorubicin, and increased IC50 from (0.05 +/- 0.00) microg/ml to (2.99 +/- 0.12) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was decreased by 30% or 35% respectively. 3. Suppression of WAVE1 in K562/A02 cells by siRNA resulted in a reversal of MDR to doxorubicin, and decreased IC50 from (4.29 +/- 0.15) microg/ml to (1.85 +/- 0.07) microg/ml, and at 1 microg/ml or 5 microg/ml of doxorubicin treatment, cell apoptotic nuclei rate was increased by 24% or 21% respectively. 4. Overexpression of WAVE1 in K562 cells significantly increased the mdrl mRNA and the Bcl-2 protein, and suppression of WAVE1 in K562/A02 cells by siRNA decreased the mRNA and the protein.

CONCLUSIONWAVE1 involves in the MDR mechanisms in K562/A02 leukemia cells through regulation the level of mdrl and Bcl-2.

Apoptosis ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Genetic Vectors ; Humans ; K562 Cells ; Plasmids ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; Transfection ; Wiskott-Aldrich Syndrome Protein Family ; genetics ; metabolism

OBJECTIVETo explore the relationship between human cyclin C (CCNC) gene and childhood acute lymphocytic leukemia (ALL).

METHODSThe total RNA isolated from myeloid tissues of normal children and of children with newly diagnosed ALL and from ALL cell line 6T-CEM was reversely transcribed into cDNA. Real-time fluorescence quantitative PCR method was used to detect CCNC gene expression.

RESULTSCCNC was expressed in myeloid tissues of normal children and of children with newly diagnosed ALL as well as 6T-CEM. The relative expression level of CCNC gene in children with newly diagnosed ALL was significantly lower than in normal controls (2.35 +/- 0.83 vs 13.5 +/- 0.30; P <0.05).

CONCLUSIONSCCNC gene shows lower expression in children with newly diagnosed ALL, suggesting that it may be a tumor suppressing gene in childhood ALL.

Child ; Cyclin C ; Cyclins ; genetics ; Female ; Fluorescence ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism

OBJECTIVETo make an attempt at the multi-element speciation in the Chinese medicinal herbs by determining the concentrations of 25 elements in different extraction solutions.

METHODFirstly, five Chinese medicinal herbs (Buddleja officinalis, Dictamnus dasycarpus, Myristica fragrans, Albizia judibrissin and Inula japonica) from the same region of China were treated to obtain water-soluble phase, lipid-soluble phase and non-soluble phase by water extraction, organic solvent extraction and acid digestion, respectively. Secondly, Phytolacca acinosa, a Chinese medicinal herb collected from 9 regions of China, was extracted by 0% EtOH, 50% EtOH, 75% EtOH, 95% EtOH, respectively, referring the Chinese Pharmacopoeia. Finally, the concentrations of 25 elements, such as Be, Cr, Cu, Zn, Ge, Sr, Y, Mo, Cd, Tl, Pb and REEs, in the above three phases were determined by ICP-MS.

RESULTUnder the optimal conditions, all the 25 elements could be determined with detection limits ranged from 0.003 to 0.71 ng x g(-1). The average recoveries of the elements in P. acinosa were 88% approximately 119%, with the relative standard deviations 1.7% approximately 13.3%. It was observed that the determined 25 elements distributed in all the water-soluble, lipid-soluble and non-soluble phases, indicating that the inorganic species, organicspecies, as well as the protein bound species were coexisted in the herbs. Big differences of the element extraction rates could be found by using different ethanol solutions.

CONCLUSIONWith the aid of the obtained results, we may increase the extraction of necessary elements while decrease that of the toxic elements from the herbs by choosing a suitable solvent during the drug production.

Buddleja ; chemistry ; Cadmium ; analysis ; Copper ; analysis ; Dictamnus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ecosystem ; Lead ; analysis ; Metals, Heavy ; analysis ; Molybdenum ; analysis ; Myristica fragrans ; chemistry ; Phytolacca ; chemistry ; Plants, Medicinal ; chemistry ; Solvents ; chemistry ; Trace Elements ; analysis ; Zinc ; analysis

Dried herb of Delphinium brunonianum Royle (Ranunculaceae) has long been used under the herbal name "Xiaguobei" (Delphinii Brunoniani Herba) in traditional Tibetan medicine and prescribed for the treatment of influenza, itchy skin rash and snake bites. In order to find a useful and convenient method for the identification of microscopic features, the technique of fluorescence microscopy was applied to authenticate "Xiaguobei" of Tibet. The transverse sections of stem and leaf, as well as the powder of "Xiaguobei" were observed to seek for typical microscopic features by normal light and fluorescence microscopy. A style-like, single-cell glandular hair containing yellow secretions on the leaf, young stem and sepal of "Xiaguobei" was found. Under the fluorescence microscope, the xylem and pericycle fiber group emitted significant fluorescence. This work indicated that fluorescence microscopy could be an useful additional method for the authentication work. Without the traditional dyeing methods, the main microscopic features could be easily found by fluorescence microscopy. The results provided reliable references for the authentication of "Xiaguobei".
Biometric Identification ; Delphinium ; anatomy & histology ; Microscopy, Fluorescence ; Plant Leaves ; anatomy & histology ; Plant Stems ; anatomy & histology ; Plants, Medicinal ; anatomy & histology ; Powders ; Tibet

OBJECTIVESTo study the clinicopathologic features of Rosai-Dorfman disease (RDD), expression of various antigens, human herpes virus type 8 (HHV8), human papillomavirus (HPV)-DNA and Epstein-Barr virus (EBV)-mRNA, and compare the findings with those in the literature.

METHODSThe clinicopathologic findings of 16 Rosai-Dorfman disease cases were retrospectively reviewed. Immunohistochemical study for S-100 protein, CD68 (PG-M1), CD163, CD21, CD1a, CD20, CD45RO, CD4, CD8, M-CSF and HHV8 was carried out in 9 of the 16 cases. In-situ hybridization for EBV-mRNA and HPV-DNA was also performed.

RESULTSThe male-to-female ratio of the patients was 4.33:1. Amongst the 16 cases studied, 62.5% (10/16) presented nodal RDD, with cervical lymph node predominantly involved. Half of these cases had affected lymph nodes in more than one anatomic site. Extranodal RDD represented 37.5% (6/16) of the cases. The relapse rate of extranodal RDD was higher than that of nodal RDD. Histologically, nodal RDD was characterized by dilated sinuses filled with large polygonal histiocytes which contained lymphocytes and plasma cells. For extranodal lesions, various degrees of stromal fibrosis were seen in association with mixed inflammatory cells (especially plasma cells). The large polygonal histiocytes varied in number and were distributed in clusters or patches. Immunohistochemical study showed that the abnormal histiocytes were strongly positive for S-100 protein. They also expressed CD68, CD163 and M-CSF, but were negative for CD1a, CD21 and HHV8. The lymphocytes in cytoplasm of these histiocytes were positive for both T and B cell markers (with T cell predominance, including a mixture of CD4- and CD8-positive cells). HPV-DNA and EBV-mRNA were not detected by in-situ hybridization. To date, 62 cases of RDD have been reported in mainland China, including 34 cases of nodal RDD and 18 cases of extranodal RDD. The remaining 10 cases involved both lymph nodes and extranodal sites. Compared with overseas reports, RDD occurring in China tended to affect older patients and with slight male predilection.

CONCLUSIONSRosai-Dorfman disease is relatively rare in China. Pathologic diagnosis of extranodal RDD may be difficult. The demographic data of RDD in China, including age and sex of patients, are different from those in the literature.

Adolescent ; Adult ; Aged ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Bone Diseases ; metabolism ; pathology ; virology ; Child ; DNA, Viral ; analysis ; Female ; Follow-Up Studies ; Herpesvirus 8, Human ; genetics ; isolation & purification ; Histiocytosis, Sinus ; metabolism ; pathology ; virology ; Humans ; Immunohistochemistry ; Lymph Nodes ; pathology ; Macrophage Colony-Stimulating Factor ; metabolism ; Male ; Middle Aged ; Nose Diseases ; metabolism ; pathology ; virology ; RNA, Viral ; analysis ; Receptors, Cell Surface ; metabolism ; Retrospective Studies ; S100 Proteins ; metabolism ; Skin Diseases ; metabolism ; pathology ; virology ; Young Adult

Objective To observe the effects of electroacupuncture at"Ciliao","Zhongji","Sanyinjiao"and"Dazhui"on urodynamics and expression of ERK/CREB/Bcl-2 pathway in spinal cord tissue of neurogenic bladder rats after suprasacral spinal cord injury.Methods Sixty female SD rats randomly selected 24 and divided into blank group and sham-operation group(12 rats in each group),the remaining 36 rats were subjected to surgical modeling.After modeling,rats were randomly divided into the model group and the electroacupuncture group,with 12 rats in each group.The electroacupuncture group received unilateral electroacupuncture stimulation at acupoints"Ciliao","Zhongji","Sanyinjiao",and"Dazhui"for 30 minutes each time,once a day,for 7 consecutive days.After administration,urodynamic testing was performed,HE staining was used to observe the morphology of bladder detrusor tissue,TUNEL method was used to detected apoptosis in spinal cord tissue,Western blot was used to detected expressions of p-ERK1/2,p-CREB,p-p90Rsk,CRE,Bcl-2,and Bax proteins in spinal cord tissue.Results Compared with the sham-operation group,the basal pressure,maximum pressure,and leakage point pressure of the bladder in the model group increased significantly(P<0.01),while the maximum capacity and compliance of the bladder decreased significantly(P<0.01);the structure of bladder smooth muscle cells was severely damaged and disorderly arranged,accompanied by a large amount of inflammatory cell infiltration;the apoptosis rate of spinal cord tissue cells significantly increased(P<0.01),and the expressions of p-ERK1/2,p-p90Rsk,p-CREB,CRE,and Bcl-2 proteins in spinal cord tissue were significantly decreased,while the expression of Bax protein significantly increased(P<0.01).Compared with the model group,the basal pressure,maximum pressure,and leakage point pressure of the bladder in the electroacupuncture group decreased significantly(P<0.05),while the maximum capacity and compliance of the bladder increased significantly(P<0.05,P<0.01);the integrity of bladder smooth muscle cells was enhanced,the degree of cell edema was reduced,and inflammatory cell infiltration was reduced;the apoptosis rate of spinal cord tissue cells was significantly reduced(P<0.05),and the expressions of p-ERK1/2,p-p90Rsk,p-CREB,CRE,and Bcl-2 proteins in spinal cord tissue significantly increased,while the expression of Bax protein was significantly decreased(P<0.05,P<0.01).Conclusion Electroacupuncture can promote the repair of bladder detrusor tissue in rats with neurogenic bladder model after suprasacral spinal cord injury,increase the maximum capacity and compliance of the bladder,alleviate the high pressure state in the bladder,and its mechanism is related to activating the ERK/CREB/Bcl-2 pathway,reducing secondary apoptosis of damaged neurons,effectively improving bladder innervation,and protecting bladder function.