1.Amplification of Full-length mRNA 5′Ends Based on Template-switching Effects
China Biotechnology 2006;0(12):-
"CapFinder" technology,which can be used to clone the full length of 5′ UTR sequence of mRNA,was described.This technology used the terminal transferase activity of certain MMLV RT variants that added 3-5 residues(predominantly dC) to the 3′end of the first-strand cDNA exhibited when MMLV RT reached the 5′cap structure of mRNA.In the reverse reaction system containing GGG oligo,the terminal transferase activity was harnessed by the GGG oligo whose terminal stretch of dG residues can anneal to the dC-rich cDNA tail and serve as an extended template for RT.After RT switch templates from the mRNA template to the GGG oligo,a complete cDNA copy of the original RNA was synthesized with the additional GGG oligo sequences at the end.5′UTR of mRNA can be amplified with GGG oligo as forward primer and a gene-specific reverse primer.5′UTR of Bt toxin receptor E-Cadherin gene in midgut of cotton bollworm was cloned.
2.Protective immunity effects of co-immunization with P30 DNA vaccine and protein vaccine
Jian SHEN ; Desheng TONG ; Yaping QIN ; Chen HUA ; Jin SI ;
Chinese Journal of Schistosomiasis Control 1989;0(03):-
Objective To study protective immunity effects of co-immunization with P30 DNA vaccine and protein vaccine. Methods Forty-eight 5-6 weeks old BALB/c female mice were divided into four groups (A,B,C,D), 12 mice of each group. In group A (control group) each mouse was immunized with 100 ?g pcDNA3.1 plasmid DNA by intramuscular (i.m.) for three times at week 0,2 and 4; in group B (P30 protein group) each mouse was immunized (i.m.) with 50 ?g rP30+50 ?g CFA for three times at week 0, 2 and 4; in group C (pcDNA3.1-P30 group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for three times at week 0, 2 and 4; in group D (P30 DNA+rP30 co-immunization group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for two times at week 0, 2 and immunized by subcutaneous with 50 ?g rP30+50 ?g CFA at week 4. Each mouse was infected with 100 tachyzoites of Toxoplasma gondii RH strain four weeks later after last immunization. The anti-P30 antibodies were detected with ELISA before the challenge. Results The P30 DNA vaccine was successfully constructed. High titers of anti-P30 antibodies were induced in each mouse immunized with DNA vaccine. The protective trial proved that there was no significant difference between control group and experimental group though the survival time of mouse from experimental group had been prolonged. Conclusion The P30 DNA vaccine could induced high titers of anti- P30 antibodies in immunized mice, and it may be a potential DNA vaccine candidate.
3.Analysis of drug resistance and clinically-isolated bacteria in neonatal infectious pneumonia of Jiaxing from 2012 to 2014
Fangfang SHEN ; Wen ZHU ; Qiaoying LU ; Jian JIN ; Yiqun TENG
Chinese Journal of Primary Medicine and Pharmacy 2016;23(8):1136-1140
Objective To evaluate the distribution and drug resistance of bacteria causing neonatal infectious pneumonia in Jiaxing,and to provide a therapy for clinical doctor to make a correct diagnosis,choose reasonable anti-biotics and avoid abuse of antibiotics.Methods Took expectoration from trachea in condition of asepsis to conduct culture and perform drug-sensitive test from 3025 cases.Results Totally 1 156 strains of aerobic bacteria were iso-lated.875 strains were gram negative bacilli(75.7%),269 strains were gram positive cocci(23.3%),and 12 strains were fungi(1.0%).Klebsiella pneumoiae,Escherichia coil,Acinetobacter baumanni,Enterobacter cloacae were com-mon in gram negative bacilli( respectively 178 cases,151 cases,87 cases,113 cases) .The proportion of the Staphylo-coccus aureus was the largest in gram positive cocci(245 cases) .The results showed that gram-negative bacilli were resistant to cefazolin, ampicillin, piperacillin and sensitive to meropenem, imipenem, piperacillin-tazuobatanna and cefoperazone-sulbactam.The drug resistance was severe of ESBL-positive.Staphylococcus aureus was resistant to penicillin, ampicillin, erythrocin, clindamycin and sensitive to linezolid, vancomycin, nitrofurantoin.Conclusion Gram-negative bacilli are the main bacteria in neonatals with infectious pneumonia.The drug resistance is severe.It is important to make a standard management and isolation.
5.Clinicopathologic analysis of primary pulmonary myxoid sarcoma with EWSR1 translocation
Yan JIN ; Xuxia SHEN ; Lei SHEN ; Yihua SUN ; Haiquan CHEN ; Jian WANG ; Yuan LI
China Oncology 2017;27(5):334-339
Background and purpose: Primary pulmonary myxoid sarcoma (PPMS) is a very rare lung tumor that has recently been shown to harbor EWSR1-CREB1 translocation. This study aimed to investigate the clinicopatho-logical characteristics and differential diagnosis of PPMS. Methods: The clinical and pathological features of 6 cases of PPMS with EWSR1 gene rearrangement were reviewed. Immunohistochemistry and fluorescence in situ hybridization (FISH) study were performed. Results: Six patients were enrolled in this study, including 4 male patients and 2 female patients with an age range of 23 to 64 years (median age, 44 years). All tumors involved pulmonary parenchyma, with a predominant endobronchial component in 1 and adjacent to bronchus in 5 patients. Microscopically, the tumor was lobulated and composed of cords of polygonal, spindle cells within myxoid stroma. Tumors were immunoreactive for only Vimentin and weakly focal for epithelial membrane antigen (EMA). All tumors were shown to harbor EWSR1 gene rearrangement by FISH. Follow-up of all patients showed that 1 patient developed a pleura and bone metastasis but was still alive and 5 were disease-free after 4-29 months. Conclusion: PPMS is an extremely rare sarcoma with low-grade malignant potential. This is characterized by distinct histological features and EWSR1 gene rearrangement. It is important that understanding tumor spectrum and genetic feature can contribute to diagnosis and differential diagnosis of PPMS.
7.Radiation dose rate received by patients with coronary heart disease during interventional management:comparison between fluoroscopy and movies
Weibin LIU ; Lianjun HUANG ; Jiufang GUO ; Yihui WANG ; Guoliang JIN ; Yujia LI ; Tao SHEN ; Bolin ZHANG ; Shen ZHANG ; Jian CHEN
Journal of Interventional Radiology 2015;(6):524-527
Objective To determine the real time radiation dose rate displayed on DSA equipment in interventional management for patients with coronary heart disease in order to provide medical staff with useful basis to reasonably control the radiation dose. Methods A total of 30 patients with coronary heart disease, who received interventional treatment at authors’ hospital in September 2014 with a GE DSA unit, were randomly selected. Intraoperative radiation dose rates when fluoroscopy was converted to movies , which were displayed on DSA at respective sequence, were determined, and the results were analyzed and compared. Results The fluoroscopy radiation dose rate ranged from 18.5 to 212.0 mGy/min, the mean value was (114.7±42.1) mGy/min;while the radiation dose rate of movies varied from 216 to 1 691 mGy/min, with a mean value of(970.1±298.4) mGy/min. The overall mean fluoroscopy-to-movies radiation dose ratio was 1 ∶8.5. The larger the digital flat panel detector area was , the bigger the mean radiation dose rate would be and the more obvious increase in the mean radiation dose rate of movies than that of the fluoroscopy would be observed; the mean radiation dose rate of fluoroscopy was lower than that of movies , and the difference was significantly. Conclusion During the performance of coronary intervention , the medical staff should make reasonable choice of the equipment and closely observe the dynamic real-time display of the radiation dose rate, promptly adjust the inspection mode and the controllable parameters when it is needed , make immediate evaluation of dose level that might cause radiation injury to the patient , and reasonably control the radiation dose to reduce the effects of ionizing radiation on human health.
8.Recurrent low frequency sensorineural deafness.
Ying LIN ; Jin Ling WANG ; Fei SUN ; Jin Jin SHEN ; Zhao Xia WANG ; Jian Hua QIU ; Ding Jun ZHA
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2018;32(6):474-476
Low frequency sensorineural deafness is a common subtype of idiopathic sudden deafness. Certain patients suffered recurrent attacks without vertigo, much alike Meniere's disease. Few of them developed into definite Meniere's disease during long-term follow-up in many clinical studies. Although the pathophysiology of recurrent low frequency deafness is yet unknown, the desease is considered associated with early state of endolymphatic hydrops or migraine. Otologists shall be aware of its clinical course and careful explanation is necessary at time of initial informed consent.
Endolymphatic Hydrops
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complications
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Hearing Loss, Sensorineural
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complications
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diagnosis
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Hearing Loss, Sudden
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Humans
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Meniere Disease
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complications
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Vertigo
9.Research of Chinese medicine pairs (VIII)--Salviae Miltiorrhizae Radix et Rhizoma-Carthami flos.
Shu-Jiao LI ; Yu-Ping TANG ; Juan SHEN ; Jian-Ping LI ; Jian-Ming GUO ; Jin-Ao DUAN
China Journal of Chinese Materia Medica 2013;38(24):4227-4231
Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos is a famous Chinese medicine pair (CMP). Salviae Miltiorrhizae Radix et Rhizoma can promote blood circulation for removing blood stasis, and Carthami Flos can promote blood circulation for removing meridian obstruction and remove blood stasis for relieving pain. The two herbs are important TCMs for activating blood. Danhong injection is the classic application of the two herbs compatibility, which was made from Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos with scientific formalation by extraction and refining. The CMP is used for treatement of organ flood insufficiency and ischemic infarction diseases. It can obviously relieving symptoms of angina pectoris, improve myocardial ischemia, regress atherosclerosis plaque, and inhibit thrombus. This paper elaberated the bio-active constituents, compatibility effects and action mechanism, and clinical applications of the CMP in order to further upgrade basic research and application levels of the CMP.
Animals
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Carthamus tinctorius
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chemistry
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Drug Compounding
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Drug Interactions
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Humans
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Medicine, Chinese Traditional
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methods
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Salvia miltiorrhiza
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chemistry
10.Early application of non-invasive ventilation in treating elderly patients with acute left ventricular failure and respiratory failure
Bao-Min FANG ; Tie-Ying SUN ; Hui-Xing KE ; Jin SHEN ; Yi-Jian QIAN ;
Chinese Journal of Emergency Medicine 2006;0(06):-
Objective To study effects of early non-invasive ventilation (NIV) application in treating elderly patients with acute left ventricular failure induced respiratory failure. Method Totally 32 elderly patients with acute left ventrieular failure induced respiratory failure, admitted from August 1997 to February 2007, received NIV treatment, and were retrospectively studied. There were 22 male and 10 female, aged (81.5?8.6) yearsdd. The changes of rahs, respiration rate, heart rate, arterial blood gas, cardiac function before and after NIV application were compared. According to the application time of NIV, 32 patients were divided into two groups: group A (early NIV application group, n=17) and group B (non-early NIV application group, n= 15). The time to improve the symptoms, the application time of NIV, cure rates, tracheal intubation rates and mortality were compared between the two groups. Results Thirty of the 32 patients survived, cardiac function was improved from New York classⅣtoⅠ~Ⅱ, respiratory rate, heart rate and blood pressure significantly decreased, PaO_2 and SaO_2 significantly increased and PaCO_2 significantly decreased. The tracheal intubation was performed in 4 patients. The time needed to improve the symptoms and the application time of NIV were significantly different between group A and group B (P