Objective To investigate the influence of gene therapy using survivin as a gene target on biological behavior of pancreatic carcinoma cell line. Methods Chemically synthesized siRNA and shRNA in pGCSi vector were used to silence survivin expression of pancreatic carcinoma cell line PaTu8988. The therapeutical effects of survivin as a gene target were evaluated through determination of the down-regulation of survivin gene expression, cellular shape, cell apoptosis, cell viability and apoptosis signal pathway changes. Results After transfection of different arrays of siRNA and shRNA vectors to silence the survivin expression, survivin mRNA and protein levels were significantly decreased (P < 0.05) ; PI staining revealed the presence of karyopyknosis, the cell apoptosis index was more than 20%; hypodiploid DNA content before G0/G1 detected by flow cytometry ; cell viability measured by MTT assay was significantly decreased (P <0.05) ; the activity of caspase-3 remarkably increased (P < 0. 05). Conclusions The pancreatic carcinoma cell line PaTu8988 be induced to promote spontaneous apoptosis procedure through silencing survivin expression by RNAi, which could accelerate carcinoma cell apoptosis and improve therapeutic effect on pancreatic carcinoma.

BACKGROUND: Ligustrazine possesses the effect to reduce ototoxicity of gentamicin, whether does it antagonize the ototoxicity of gentamicin through influencing the expression of heat shock protein (HSP) 70 in cochlea of guinea pigs with gentamicin induced ototoxicity? BJECTIVE: To investigate the effect of ligustrazine on the expression of HSP70 in cochlea of guinea pigs with gentamicin induced ototoxicity through immunohistochemistry and image analysis technique in combination with the measurement of auditory brainstem response (ABR).DESIGN: A randomized controlled study, and linear correlation analysis. ETTING: Physiological Department of Shenyang Medical College and Audiological Laboratory of China Medical University.MATERIALS: Totally 40 white and red-eye healthy guinea pigs of lean grade and with keen auricle reflect, weighing 200 - 250 g, of either gender, were at random divided as gentamicin group, ligustrazine + gentamicin group, ligustrazine group, and normal control group, with 10 in each group.METHODS: Ligustrazine injection 140 mg/kg was intraperitoneally at the left side given for animals in ligustrazine + gentamicin group, and at the same time gentamicin sulfate injection 100 mg/kg was given intraperitoneally at the right side. The same dosage of gentamicin sulfate injection was intraperitoneally given for animals in gentamicin group. The same dosage of ligustrazine was intraperitoneally give for animals in ligustrazine group. The normal saline 2.5 mL/kg was intraperitoneally given for animals in normal control group. Administration was consecutively given for 10 days for each group, once a day. The body mass was measured every day for regulating the dosage. Before starting and after finishing the administration, the thresholds of ABR of all animals in each group were measured respectively. After finishing administration, all animals in each group were put to death, and the conditions of expressions of HSP70 in cochlea of guinea pigs were investigated by SABC immunohistochemistry and image analysis technology.MAIN OUTCOME MEASURES: ① the thresholds of ABR of all animals in each group; ② expressions of HSP70 in cochlea of guinea pigs in each group.RESULTS: ① The thresholds of ABR of guinea pigs: The thresholds in ligustrazine + gentamicin group and gentamicin group were obviously higher than that in normal control group [(21.09±4.50) dBnHL, (36.55±6.13)dBnHL, (2.50±2.75) dBnHL, t=15.764-22.665, P ＜ 0.001]. The threshold in ligustrazine + gentamicin group was obviously lower than that in gentamicin group (t=9.092, P ＜ 0.001). ② The expressions of HSP70 in each part of cochlea of guinea pigs: The mean gray values of cellular HSP70 positive reaction products in Corti's organ, vascular stria and spiral ligament, spiral limbus and spiral ganglion in gentamicin group were lower than those in normal control group (88.24±4.34, 96.85±1.05; 121.24±0.92,128.76±1.59; 96.15±1.10, 98.78±0.54; 117.73±1.18, 120.51±0.80, t=6.097-18.307, P ＜ 0.001). The mean gray values of cellular HSP70 positive reaction products in ligustrazine + gentamicin group were lower than those in gentamicin group (92.53±2.25, 88.24±4.34; 125.20±1.43, 121.24±0.92;98.71±0.91, 96.15±1.10, 118.91±0.46, 117.73±1.18, t=3.925-10.415, P＜ 0.001).③ The mean gray value of cellular HSP70 positive reaction products in each part of cochlea of guinea pigs was highly correlated with the threshold of ABR (r=-0.814 1 to -0.984 1, P ＜ 0.001).CONCLUSION: Ligustrazine could reduce the threshold of ABR and the expression of HSP70 in cochlea with gentamicin toxicosis so as to relieve gentamicin ototoxic injury, hence improving auditory function.

Thirteen compounds were isolated from the ethyl acetate fraction of Crepis crocea by column chromatographies on silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures were elucidated on the basis of spectral analysis as tectorone I (1), 8β- (2-methyl- 2-hydroxy-3-oxobutanoyloxy) -glucozaluzanin C (2), tectoroside (3), luteolin-7-O-glucoside (4), cosmosiin (5), esculetin (6), 3,4-dihydroxybenzaldehyde (7), trans-4-hydroxycinnamic acid (8), Caffeic acid (9), methyl p-hydroxyphenyllactate (10), ethylp- hydroxyphenyllactate (11), cis-3,4-dihydroxy-β-ionion (12). All the compounds, except for compounds 4 and 9, were isolated from this plant for the first time, and tectorone I (1) is a new natural product.
Crepis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure

Objective To investigate the role of interleukin 17 in the differentiation of macrophages from monocyte in vivo and its applicability in Obliterative bronchiolitis (OB).Methods Pathogen-free male C57BL/6 (n =20),IL-17-defcient C57BL/6 (n =10) and BALB/c (n =10) weighing 20-25 g.Weight-matched mice were assigned to three experimental groups.Experimental group A:BALB/c (n =5) →C57 BL/6 (n =10).Experimental group B:BALB/c (n =5) →IL-17-defcient C57BL/6(n =10).Normal control group:C57BL/6(n =10).The CD80 expression in macrophages in spleens were analysed by flowcytometry 7 days after operation.Native and transplanted lungs were harvested after 14 days and 28 days,stained with HE,and examined under light microscopy.Results Compared to experimental group A,CD80 expression of macrophages reveales a significant decrease in spleen of experimental group B.Furthermore,airway obliteration and destruction of the epithelium in experimental group A were significantly better compared with that of experimental group B on day 14 and 28 after transplantation.Conclusion IL-17 deficiency decreased the polarization of monocytes to macrophages and attenuate the pathology of obliteral bronchiolitis in murine model after trachea transplantaion.

OBJECTIVETo observe the clinical efficacy of Zengse Pill ( ZSP) on patients with vitiligo of qi-stagnancy and blood-stasis syndrome type (V-QB), and to preliminarily explore its mechanism of action.

METHODSSixty-five V-QB patients, with their diagnosis confirmed by clinical examination, were randomized by digital table method into two groups, with 31 patients in the control group and 34 in the treatment group. Cobamamide (2 tablets) was administered orally to all patients, and Psoralea tincture (a self-formulated preparation) was applied externally thrice a day. In addition, for patients in the treatment group, ZSP was given orally, at 5 pills per dose, 3 times every day. The therapeutic course for both groups was 3 months. Patients were re-examined every half-month, and changes in the skin lesions were observed and recorded. The levels of lymphocyte subsets, serum immune globulin, and complement C3 and C4 in patients were determined before and after the therapeutic course and compared with the corresponding indexes determined in 21 healthy subjects.

RESULTSThe total effective rate in the treatment group was 82.4%, which was markedly higher than that in the control group (54.8%), showing a significant difference (P<0.05). After treatment, CD(4) (+) percentage, CD(4) (+)/CD(8) (+) ratio, and blood levels of C3 and C4 increased, while CD(8) (+) percentage decreased in the treatment group (P<0.05 or P<0.01). All these indexes remained unchanged in the control group, and the respective comparisons between groups showed significant differences (P<0.01).

CONCLUSIONZSP has a definite clinical effect on the treatment of V-QB but with no evident adverse reactions, and it can increase the CD(4) (+) percentage, CD(4) (+)/CD(8) (+) ratio, and the levels of serum C3 and C4, thus regulating the immunity of the organism, which might be one of its mechanisms of action.

Adolescent ; Adult ; CD4-CD8 Ratio ; Complement C3 ; analysis ; Complement C4 ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulins ; blood ; Lymphocyte Subsets ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; Vitiligo ; drug therapy ; immunology

OBJECTIVETo investigate the effects of cyclosporine A (CsA), a powerful inhibitor of mitochondrial permeability transition pore (MPTP), on pneumocyte apoptosis, the release of cytochrome C and the activity of caspase-3 after lung ischemia/reperfusion, and explore the mechanisms.

METHODSSingle lung in situ ischemia/reperfusion animal model was used. 30 SD rats were randomly divided into three groups (n = 10): sham (S) group, ischemia/reperfusion (I/R) group and cyclosporine A (CsA) group. Apoptosis of pneumocyte was assessed by TUNEL method, cytochrome C (CytC) in cytoplasm was detected by immunohistochemistry techniques, and the activity of caspase-3 was measured with spectrophotometer.

RESULTSThe content of CytC in cytoplasm, the activity of caspase-3, and the value of apoptosis index (AI) in ischemia/reperfusion group were evidently higher than that in S group (P < 0.01). CsA suppressed apoptosis as well as CytC release and caspase-3 activity (P < 0.01).

CONCLUSIONCsA can prevent the release of cytochrome C, block the apoptosis of pneumocyte accordingly maybe by closing the MPTP.

Alveolar Epithelial Cells ; cytology ; drug effects ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cyclosporine ; pharmacology ; Cytochromes c ; metabolism ; Lung ; blood supply ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology

This paper introduces the old nine-needle including its name, shape, function, manipulation and origination, as well as systematically introduces the new nine-needle as a new product which has been developed under the guidance of modern science and technology based on the old nine-needle since 1949. As a new tool, the new nine-needle provide much more choice for acupuncture treatment and obtained satisfied clinical effect. During the development of research and clinical practice on the new nine-needle, its theory has been consummated successively. Combining various kinds of needles for treatment is not only paying attention to the specificity of one kind of needles, but also emphasizing the whole function of all kinds of needles, which makes the new nine-needle formed the characteristic and distinct theory of its own.
Acupuncture Therapy ; history ; instrumentation ; methods ; China ; History, Ancient ; Needles

With the unique structure and manipulation techniques of blood-letting and cutting, hook needle serves as a role in dredging meridians, removing blood stasis, purging heat for resuscitation and relaxing synechial tissues. There are three needling techniques of hook needle: swift pricking, bleeding and pricking. Six manipulations are commonly used as lifting-thrusting, plucking, pulling, pushing-scraping, vibration and massage. The mechanism and manipulations of hook needling are summarized systematically in the present paper. In this way, its manipulations, indications and contraindications are expounded in detail.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Needles

Objective: To investigate the clinicopathological, and molecular characteristics of myoepithelial tumors (MTs) of salivary glands. Methods: A total of 37 MTs cases including 13 malignant epithelial tumors (MMTs) and 24 benign epithelial tumors (BMTs) of salivary glands were identified from the archives of the Department of Pathology, General Hospital of Eastern Theater Command, dating from 2006 to 2016. Clinical features, histological patterns, immunohistochemical characteristics and status of EWSR1 gene rearrangement by fluorescence in situ hybridization (FISH) analysis were reviewed in all cases. Results: Clinically, 37 MTs cases mainly occurred in the parotid glands, when most of the patients presented with painless masses. Of the 13 MMTs cases, male to female ratio was 7∶6, and the median age was 62 years old. Of the 24 BMTs cases, male to female ratio was 5∶7, and the median age was 54 years old. Immunohistochemically, 37 MTs cases were positive for CKpan, and at least one myoepithelial marker. Twenty six of 37 MTs cases were analyzable for the EWSR1 gene break by FISH. Based on the previous evaluation criterion, the EWSR1 translocation was detected in 4 cases of 11 MMTs, and 4 cases of 15 BMTs. According to the main histological composition of tumor cells, 4 EWSR1-positive MMTs covered 2 clear-cell cases and 2 epithelioid-cell cases, when 4 EWSR1-positive BMTs covered 2 clear-cell cases, 1 plasmacytoid-cell case, and 1 spindle-cell case. Conclusions Males and females are affected equally. MTs express immunoreactivity for CKpan, and at least one myoepithelial marker. The EWSR1 rearrangement is present in a subset of MTs, with variable morphological characteristics, and has no statistical significance on clinical behavior.

Objective: To investigate the clinicopathological features of mammary analogue secretory carcinoma (MASC) of salivary glands, and its diagnosis, differential diagnosis, immunohistochemistry and molecular pathology. Methods: Seventeen cases of MASC were enrolled, with 9 cases of salivary acinar cell carcinoma and 18 cases of adenoid cystic carcinoma as control groups from Nanjing General Hospital from 1997 to 2014 were included in this retrospective study, combined with immunohistochemistry and molecular detection of ETV6-NTRK3 gene fusion. All cases were histologically reviewed with immunohistochemical staining (EnVision) for S-100 protein, SOX10, GATA3, CD117 expression in each group. Fluorescence in situ hybridization (FISH) was used to detect the ETV6-NTRK3 gene fusion. Results: The age of MASC patients ranged from 27 to 74 years with mean age of 47 and ratio of male and female was 4∶3. All cases showed infiltrative growth and diverse cytology and histology, including lobular (8 cases), cystic papillary (3 cases), cribriform mixed with papillary and glandular structures (6 cases) at various proportions. Some tumors of MASC also exhibited solid growth areas with occasional microcystic honeycombed pattern composed of small cysts merged into larger cysts resembling thyroid follicles. S-100 protein and SOX10 were strongly positive in all MASC cases (17/17). In addition, there was insignificant positivity for GATA3 (3/17) and CD117 (4/17). ETV6 gene fusion detection was informative in 12 MASC cases by FISH with 10 positive cases and 2 negative cases. Conclusions Combined immunohistochemical positivity of S-100 protein, CD117 and SOX10 are useful in the diagnosis and differential diagnosis of MASC. FISH detection of ETV6-NTRK3 fusion offers an additional molecular diagnostic marker for the diagnosis.