Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Combined Modality Therapy ; Female ; Humans ; Male ; Manipulation, Orthopedic ; Middle Aged ; Shoulder Dislocation ; therapy ; Young Adult

The operation and application of Tiantu (CV 22) in emergency are discussed in the paper. The stimulating methods at Tiantu (CV 22) are acupuncture and pressing technique. The correct insertion of needle and proper depth and direction of insertion are required during acupuncture. The pressing technique stress the pressing strength and pressing time. Acupuncture or pressing technique is suitable for the emergent measurement of asthma, asthmatic breathing, coma, blockage of phlegm, hiccup, sore throat, etc. It is indicated that Tiantu (CV 22) is the key point in the emergency and phlegm resolving. Based on the characteristics of the point as promoting qi circulation, reducing the reversed qi and resolving phlegm, in light of the proper points combination by different syndromes and in terms of the correct and safe stimulating methods, Tiantu (CV 22) can achieve the immediate therapeutic effects in the emergent situations.
Acupuncture Points ; Acupuncture Therapy ; Asthma ; therapy ; Emergency Medicine ; Humans ; Qi

OBJECTIVETo evaluate the role of 1H spetral selected point-resolved spectroscopy (SS-PRESS) sequence in distinguishing benign from malignant breast lesions by the malignancy marker of choline peak and to investigate the factors influencing the diagnosis.

METHODSA total of 131 patients (aged 24-83 years, average 44.8 years) were enrolled in this study. The examinations were performed on a 1. 5T scanner with four-channel phased array breast coil. Single-voxel proton magnetic resonance spectroscopy (1H MRS) was acquired by SS-PRESS sequence in these patients referred to surgical or biopsy consultation.

RESULTSAmong these patients, 74 were proved to have breast carcinomas and 57 have benign lesions by histopathological examinations. Thirty-one elevated choline peaks were observed in these 74 confirmed malignant lesions, and 5 detectable choline peaks were demonstrated in the 57 benign lesions. The sensitivity and specificity of 1H SS-PRESS MRS were 41.9% and 91.2%, respectively. The main factors influencing the diagnosis were signal-to-noise ratio and pathological type.

CONCLUSIONS1H SS-PRESS MRS can provide a noninvasive, biochemical measurement of metabolism and improve the specificity of breast magnetic resonance imaging. Choline peak in vivo is a specific but not sensitive marker of malignancy. Technique factors and histopathological characterization of lesions influence the detection rate.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; diagnosis ; pathology ; Carcinoma, Ductal, Breast ; diagnosis ; pathology ; Choline ; Female ; Fibroadenoma ; diagnosis ; pathology ; Fibrocystic Breast Disease ; diagnosis ; pathology ; Humans ; Image Enhancement ; Lymphatic Metastasis ; Magnetic Resonance Imaging ; methods ; Magnetic Resonance Spectroscopy ; methods ; Middle Aged ; Sensitivity and Specificity

OBJECTIVETo establish the method for cotransferring human A20 gene and human heme oxygenase-1 (HO-1) gene into the isolated rat islets using lentiviral transfection system, and to study the protective effect of A20 and HO-1 protein against the apoptosis induced by cycloheximide (CHX) and TNF-α, and finally to explore the underlying mechanism.

METHODThe A20 gene and HO-1 gene were cloned and inserted into the lentiviral transfection system. The efficacy of gene transfer was measured by the intensity of the enhanced green fluorescent protein (EGFP) fluorescence-positive islets. Western blot was applied to verify the expression of the A20 and HO-1 genes. To induce apoptosis in vitro, the isolated islets were treated with CHX+TNF-α, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and the fluorescence-activated cell sorting (FACS) methods were used to evaluate the apoptosis of the islet cells and Western blot was used to detect caspase-3 activation.

RESULT(1) A20 and HO-1 genes were introduced into the isolated islets by lentiviral transfection, both of the genes were highly expressed in the islets after 96 hours culture detected by Western blot method. (2) The insulin levels in the cell culture medium from A20 and/or HO-1 transgenic islets were significantly higher than that in non-transgenic controls (P < 0.01). (3)After CHX + TNF-alpha treatment, the cell culture medium insulin concentration in the A20 gene transfected group [(93.58 ± 4.12)µg/ml], HO-1 gene transfected group [(88.98 ± 4.77) µg/ml ] and A20/HO-1 co-transfected group [(103.33 ± 3.16) µg/ml] were significantly higher than that in the EGFP group [(9.03 ± 0.65) µg/ml ] and the control group [(8.86 ± 0.38) µg/ml] (P < 0.001). Minimum expression level of the activated caspase-3 was found in the A20/HO-1 co-transfected group.

CONCLUSIONThe lentiviral gene transfer system was an efficient and stable gene transfer vector, the over-expressed A20 and HO-1 protein delivered via lentivirus could preserve rats' islets function and act against the apoptosis induced by CHX and TNF-α.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Flow Cytometry ; Genetic Vectors ; Heme Oxygenase-1 ; genetics ; metabolism ; Humans ; Insulin ; metabolism ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Islets of Langerhans ; drug effects ; enzymology ; physiology ; Lentivirus ; genetics ; Male ; Nuclear Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection ; methods ; Tumor Necrosis Factor alpha-Induced Protein 3 ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo develop a simple, rapid and reliable method of purifying Sprague-Dawley (SD) rat islets by sequential filtration through two cell strainers of different sizes and to evaluate the efficacy of the method.

METHODSIslets were isolated from 8 to 12-week-old clean grade Sprague-Dawley rat pancreases using the standard collagenase digestion procedure and purified with either the generally used Ficoll density gradient method or the innovative two-step sequential filtration method. The purity and vitality of the isolated islets were visualized and assessed with DTZ and AO/PI staining. Glucose stimulating tests were performed to assay cell activity, and immunohistochemical staining was used to evaluate the synthesis function of islet cells.

RESULTSThe yield of islets in the two-step filtration method group was 782±115 IEQ per rat, which was significantly higher than in the conventional Ficoll density gradient method group (598 ± 135 IEQ per rat, P < 0.01). Purity of the isolated islets in the two-step filtration method group was 90%-100% and vitality was over 95%. In the conventional Ficoll density gradient method group, islet purity was 65%-85% and vitality was 85%-95%. With regard to the high-sugar stimulation test in the two-step filtration method group, insulin concentrations in islets cultured for 24 hours were significantly higher than in those that were freshly purified (76.9 ± 6.1 μg/L vs 49.4 ± 3.9 μg/L; P < 0.01).

CONCLUSIONSA two-step sequential filtration method for rat islet purification was developed and the method was simple and reliable, with high islet vitality, purity and yield.

Animals ; Cell Separation ; methods ; Female ; Filtration ; Immunohistochemistry ; Insulin ; biosynthesis ; Islets of Langerhans ; cytology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the clinicopathological significance of positive CK7 expression in human colorectal carcinoma (HCC).

METHODSImmunohistochemistry was used to detect CK7 and CK20 protein expressions in 68 cases of HCC, 20 cases of canalicular adenoma (CA), 5 cases of serrated adenoma (SA) and 20 cases of hyperplastic polyps (HP).

RESULTSThe positivity rate of CK20 expression was 89.7% in HCC, and 100% in CA, SA and HP. In HCC, the expression rate of CK7 (39.7%) was not correlated with Dukes' classification, differentiation and tumor location. CK7 positivity rate in colon cancer was 35.7% (15/52) and 42.3% (11/26) in rectal cancer. CK7 expression was negative in CA. CK7 positivity rate in SA was 49% and 30% in HP.

CONCLUSIONCK7 is a possible marker for colorectal carcinogenesis from HP to SA, and ultimately to HCC, and examination of the colorectal polypoid lesions for CK7 expression can be significant for estimating the colorectal polypous cancerization.

Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cytoplasm ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; metabolism ; pathology ; Keratin-7 ; metabolism ; Male ; Middle Aged

OBJECTIVETo observe the effects of antisense RNA of connective tissue growth factor (CTGF) on rat liver fibrosis.

METHODSGene recombinant techniques were used to construct a rat antisense RNA of CTGF recombinant plasmid which could be expressed in eukaryotic cells. The recombinant plasmids were encapsulated with lipofectamine and then transducted into a carbon tetrachloride (CCl4) induced rat liver fibrosis model. Expression of CTGF was assessed by RT-PCR, Western blot and immunohistochemistry. Immunohistochemistry was used to identify type I and III collagens. HE stained liver slides were used for pathological study.

RESULTSThe mRNA and protein expression of CTGF in the fibrotic liver transfected with antisense-CTGF were significantly decreased compared with those of the controls (P<0.01). The depositions of type I and type III collagens were also decreased (P<0.05). Antisense-CTGF also minimized the pathological fibrosis in the rat livers (P<0.01).

CONCLUSIONThe results demonstrate that the antisense RNA of CTGF recombinant plasmid has certain effects in preventing liver fibrosis and makes it a possible candidate for use in future gene therapy.

Animals ; Connective Tissue Growth Factor ; genetics ; Genetic Therapy ; Liver ; pathology ; Liver Cirrhosis, Experimental ; pathology ; Male ; Plasmids ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

Objective To analyze the epidemiology,infection status,risk factors and microbiological characteristics of Trichosporon asahii in urinary tract infection for guidance of selecting the prompt and effective antifungal drugs in clinical therapy.Methods A total of 18 strains of Trichosporon asahii isolated from the patients with urinary tract infection were selected from 2013 to 2016.The isolation and identification of pathogenic bacteria,results of antimicrobial susceptibility test and clinical data were investigated by retrospective epidemiological survey.Results The 5 antifungal drugs,i.e.,5-fluorocytosine,amphotericin B,fluconazole,itraconazole and voriconazole,exhibited favorable antibacterial activity for the 18 strains of Trichosporon asahii with resistance rate of 0,5.6％,0,0 and 0 except itraconazole which showed only 50％ of sensitive rate.The risk factors of Trichosporon asahii infection in urinary system mainly included such as male,basic diseases (100％),long-term use of broad-spectrum antimicrobial agents (100％),indwelling catheter (83.3 ％),application of corticosteroids (50.0％) and immunosuppressive agents (38.9％) as well as a small proportion of granulocytopenia (5.6％).The 16 cases treated with fluconazole were improved,while the other 2 cases died following the treatment with itraconazole or voriconazole for reasons irrelevant to antifungal treatment.Conclusion Trichosporon asahii could cause urinary tract infections with high risk factors including basic diseases,long-term use of broad-spectrum antimicrobial agents,indwelling catheter,etc.The drug of top choice should be fluconazole.The key elements for successful treatment of Trichosporon asahii infection include early diagnosis of pathogens and correct selection of antifungal agents based on sensitivity and resistance tests of drugs.

Objective: To explore the application value of endoscope in probing the chronic wound with sinus tract in clinic. Methods: Twenty-eight chronic wounds with sinus tracts from 27 patients conforming to the inclusion criteria admitted to Outpatient Department of Wound Healing Center of Ruijin Hospital from December 2017 to March 2018 were investigated in a prospective and self-controlled trial. After being cleaned, the diameter of the opening of sinus tract was measured with a rule. A probe was used to measure the depth of a sinus tract according to the touch from the probe extremity in operation, and to measure the depth of a sinus tract that could be observed with naked eyes with the help of a pair of hemostatic forceps. Five minutes later, a probe was inserted deeply into the sinus tract to measure the depth under the endoscopic view combined with touch from the probe extremity in operation. Afterwards, the sinus tract was observed with endoscope, and the depth of the tract which could be observed under the endoscopic view was measured using a probe inserted deeply into the sinus tract. After completion of the above exploration, the sinus tract was infused with contrast agent Omnipaque 350 and scanned by computed tomography (CT) later to obtain its depth. The following indicators were calculated: the ratio of the depth of the sinus tract measured by CT to the diameter of the opening of the sinus tract (hereinafter referred to as the depth/diameter ratio of the sinus tract), the deviation rate comparing the depth of the sinus tract measured by conventional method (measured by probe only) and by endoscope (measured by probe under the endoscope view) with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the measured depth of the sinus tract), the deviation rate comparing the depth of the sinus tract that could be observed measured by conventional method and by endoscope with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the depth of the sinus tract that could be observed). Data were processed with paired t test. Pearson correlation analysis was applied to analyze the correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope. Results: The depth/diameter ratio of the sinus tract of this group of wounds was 1-32 (8±7). The deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method were (19±14)% and (79±18)%, respectively, both obviously larger than (9±9)% and (25±25)% by endoscope (t=3.837, 13.626, P<0.01). Positive correlation existed between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by conventional method, and between the depth/diameter ratio of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope (r=0.514, 0.585, 0.651, P<0.01). However, there was no obvious correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by endoscope (r=0.113, P>0.05). Conclusions Compared with the conventional method, application of endoscope is able to get more accurate data of chronic wounds with sinus tracts and observe the wounds with wider range.

This study was purposed to constructe the three-plasmid system of the lentiviral vector carrying the green fluorescent protein (GFP) gene and to investigate the expression of GFP in T lymphocytes of the mouse. The polypurine tract (PPT) element, ubiquinone promoter (PUB) and GFP were ligated to plasmid pLO134 using subcloning technology to construct plasmid pTK153. Human kidney 293T cells were co-transfected with the three-plasmid system containing packaging plasmid DeltaNRF, plasmid pTK153 and envelope plasmid VSV-G by using calcium phosphate DNA precipation and the expression of GFP was observed under fluorescence microscope after 12 hours. The viral particles were collected after transfection 72 hours, were frozen at -80 degrees C and were used to infect mouse T lymphocytes at multiplicity of infection (m.o.i.) of 3. The expression of GFP in mouse T lymphocytes was observed by fluorescence microscopy and fluorescence-activated cell sorting (FACS). The results showed that the transfection efficacy was 63.04 +/- 7.24% in 293T cells analysed by FACS and the viral titer was (3.09 +/- 0.61) x 10(6) U/ml. The expression of GFP was also evident in mouse T lymphocytes and the transduction efficacy was (37.98 +/- 6.26)%. It is concluded that the three-plasmid system of lentiviral vector containing GFP gene is successfully constructed and the transduction efficacy is high in mouse T lymphocytes.
Animals ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Viral ; analysis ; T-Lymphocytes ; metabolism ; Transduction, Genetic