Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.
Codon ; Escherichia coli ; Genome, Fungal ; Reishi ; genetics ; Saccharomyces cerevisiae ; Transcriptome

To establish HPLC specific chromatogram and its correlation with the protection effect of Shuanghuanglian on MDCK (Madin-Darby canine kidney) cell injury induced by influenza A virus( H1N1). Nine recipes of Shuanghuanglian based on the official prescription were prepared according to orthogonal test for HPLC analysis and MDCK cells protection experiment separately (cytopathic effect (CPE) method was used for observing the virus infectivity and MTT staining results were used as the determining indexes for drug concentration selection and analyzing cell viability). The results suggested that all the other Shuang-Huang-Lian recipes except recipe1 demonstrate protecting effect on MDCK cell injury induced by influenza A virus (P < 0.01, P < 0.001). Stepwise regression analysis was used for analyzing the relationships between HPLC fingerprint and the protecting effect of Shuanghuanglian on influenza A virus induced MDCK cell injury. Peak 2, 3, 6, 8 and 12 were found to be strongly related with anti-influenza A virus efficacy. Stepwise regression analysis of recipes data and efficacy data showed that Lonicerae Japonicae Flos and Forsythiae Fructus were positively associated with the protecting effect of cells injury. From HPLC fingerprints, we found that peak 2, 3, 12 were from Lonicerae Japonicae Flos and peak 6, 8 were from Forsythiae Fructus. Four peaks were identified through comparing the retention time between the standard and Shuanghuanglian recipes, and they were chlorogenicacid, cryptochlorogenic acid, forsythoside B and 3,4-dicaffeoylquinic acid respectively. Caffeic acid derivatives in Lonicerae Japonicae Flos and Forsythiae Fructus were found to be greatly correlated with anti-influenza A virus efficacy and maybe the substance basis of Shuanghuanglian.
Animals ; Antiviral Agents ; analysis ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Dogs ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Forsythia ; chemistry ; Influenza A Virus, H1N1 Subtype ; drug effects ; physiology ; Lonicera ; chemistry ; Madin Darby Canine Kidney Cells ; Scutellaria baicalensis ; chemistry

OBJECTIVETo investigate the effect and mechanism of Bufei Qingyu Granule (BQG) in mollifying the skin of scleroderma model mice.

METHODSScleroderma model induced with bleomycin in BALB/C mice 8-weeks old were administered with different dose of BQG for 26 days. The pathological changes of the mice skin were observed.

RESULTSTreatment with low, medium and high dose of BQG showed a tendency to ameliorate the thickened dermis in scleroderma mice but without statistical significance. Medium and high dose of BQG reduced the perivasculitis of dermis and alleviated the reduction or deletion of accessory structure, such as hair follicle and sweat gland. And the spleen index was lower markedly in mice treated with BQG of any dose than that in the untreated model mice (P < 0.05).

CONCLUSIONBQG could ameliorate the sclerosed skin in model mice and prevent the occurrence of splenomegaly.

Animals ; Bleomycin ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Phytotherapy ; Random Allocation ; Scleroderma, Localized ; chemically induced ; drug therapy ; Skin ; drug effects ; pathology ; Time Factors

To determine the effects of Von Hippel-Lindau (VHL) on the invasion and migration of glioma U251 cells. Methods:U251 GBM cells were transfected using VHL expression plasmid. Real-time polymerase chain reaction was conducted to de-tect VHL mRNA expression after transfection. Western blot assay was used to measure protein (VHL, MMP-2, and MMP-9) expres-sion. Tumor invasion and migration were examined by the Transwell and wound-healing experimental methods after VHL up-regula-tion. The intracranial model of nude mouse was developed using U251 cells transfected by VHL expression plasmid, and immunohisto-chemical staining was used to measure protein (VHL, MMP-2, and MMP-9) expression in the tissue sections. Results: In the U251 cells transfected by VHL expression plasmid, the expression of VHL mRNA and VHL proteins increased, and the expression of MMP-2 and MMP-9 protein decreased. Meanwhile, the invasion and migration of glioma U251 cells were also inhibited. Immunohistochemical staining results showed that the expression of MMP-2 and MMP-9 proteins decreased, and the VHL protein expression increased after transfection. Conclusion:VHL can inhibit the invasion and migration of glioma U251 cells. Thus, VHL gene can be used as a target for the gene therapy of gliomas.

Background and purpose: Short tandem repeats (STR) multiplex PCR fluorescence detection technology is the most widely used DNA technology in individual identity and genetic identification. It's the most direct method to obtain accurate conclusions. However, some studies have indicated that the rate of STR mutations in tumor tissue is significantly higher than that in normal tissues or blood. This study aimed to investigate the tendency of genetic instability in 20 STR loci on autosomal and Amel loci in tumor tissue samples from lung cancer. Methods: This study, collected 75 cases of human lung cancer tissues and the adjacent normal tissues. DNA samples were extracted by tissue DNA extraction kit, amplified using MicroreaderTM 21 Direct ID System PCR amplification kit. Capillary electrophoresis was performed using API 3130 analyzer, and results were analyzed by genetic analysis software (Gene Mapper ID V3.2). Results: STR alterations were detected in 24 specimens from 75 lung cancer tissues (32%). Fifty-five alterations were detected in the frequently used 21 STR loci in total, including additional alleles 10 times, loss of heterozygosity 10 times, partial loss of heterozygosity 35 times. Partial loss of heterozygosity was the most common genetic alteration types accounting for 63.64% of the total alteration frequency. And multiple genetic alteration types could occur in the same lung cancer tissue. Among them, the highest alteration frequency occurred on D5S818 (7 times), secondly on D3S1358 and D12S391 (both 5 times), and no alterations on D2S441 and Penta E. Combining the experimental results and analysis on clinical data, this study found the statistical differences between the staging of lung cancer and the age of the patients with the STR loci alterations (P<0.05). However, the alterations did have much relationship with the classification of lung cancer and the patient's gender (P>0.05). Conclusion: STR loci of the lung cancer tissue were not stable, and the alteration occurred in the aged or high malignant degree lung cancer tissue more frequently. Meanwhile, no alteration was detected on D2S441 and Penta E. In the future research the two STR loci should be verified to determine whether they can be used as the stable STR loci in such cases by increasing the sample size.

Objective To investigate the value of high-dose dobutamine stress echocardiography combined with two-dimensional strain imaging in early diagnosis of coronary artery disease. Methods Highdose dobutamine stress echocardiography was performed to 28 patients with suspected coronary artery disease. All wall movements were observed during resting condition and at all stress levels,respectively;the peak systolic longitudinal strain in each endomyocardial segment of left ventricular was measured; the sensitivity and specificity between visual method and two-dimensional strain imaging in diagnosing myocardial ischemia with high-dose dobutamine stress echocardiography were compared. The average peak systolic longitudinal strain was calculated against control group, coronary artery disease group during ischemia segments and non-ischemia segments, and a comparison was made inside each group as well as against the other groups. The area under receiver operating characteristic curve of the peak systolic longitudinal strain was used to predict the sensitivity and the specificity of myocardial ischemia. Results With dobutamine dose of 40 μg·kg-1 · min-1 ,wall motion abnormalities were diagnosed in 6 patients (20 segments) through visual method, myocardial ischemia was found in 15 patients (148 segments) through computing the peak systolic longitudinal strain. Inside the coronary artery disease group during ischemic　segments,the majority of peak systolic longitudinal strain was significantly reduced ( P＜0.05) compared to the non-ischemic segments and the control group. In diagnosing myocardial ischemia in high-dose dobutamine stress echocardiography, the sensitivity of visual method and two-dimensional strain imaging were 35.3％ and 88.2％(P＜0.01), specificity 100％ and 100％(P＞0.05), and accuracy 60.7％ and 92.8％ (P＜0.01). The cutoff value of the peak systolic longitudinal strain was less than or equal to 14.9％, its sensitivity and specificity in predicting myocardial ischemia were 83.3％ and 91.7％,respectively. Conclusions High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can increase the sensitivity of detecting myocardial ischemia and detect concealed myocardial ischemia. High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can be used in early diagnosis of coronary artery disease.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
Anti-Inflammatory Agents ; pharmacology ; Carthamus tinctorius ; Computational Biology ; Drugs, Chinese Herbal ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Humans ; Inflammation ; drug therapy ; Medicine, Chinese Traditional

Clinical research design is the beginning and the cornerstone for clinical research,and it functions as the preliminary step of translational research.In order to improve clinical research ability,foster quality research talents team,and to promote the development of translational research,Peking Union Medical College Hospital (PUMCH) started to provide the online course Designing Clinical Research (DCR),which has greatly contributed to enhancing clinical research capacity.The management of DCR online course serves as a great exploration on training junior medical professionals and international research collaboration.This study aims at sharing the management experiences of clinical research design and junior talents training.

Many drug candidates identified from natural products are poorly water-soluble. The surfactants used to disperse the hydrophobic anticancer drugs in water may cause a serious of acute hypersensitivity reactions. Nanotech?nology provides an alternative strategy for delivery of anticancer drugs. Drugs can be encapsulated or attached to the nanomaterials such as lipids, polymers and solid-core nanoparticles. In the present study, porous inorganic nanoparti?cles have been utilized for delivery of water-insoluble anticancer drugs. The synthesized nanoparticles were functional?ized with different organic polymers. The porous nanoparticles were readily internalized by human glioblastoma U-87 MG cells, and didn't display cytotoxicity. The internalized nanoparticles were mainly localized in endosomes/lysosomes in cells. With the hydrophobic curcumin and carfilzomib as model drugs, intracellular delivery of hydrophobic anticancer drugs by the porous inorganic nanoparticles was studied. The porous nanoparticle-based encapsulation of hydrophobic drug provides the aqueous dispersion of the drugs. In endosomes/lysosomes mimicking buffers with a pH of 4.5-5.5, pH-dependent drug release was observed from drug loaded nanoparticles. The intracellular drug content and cytotoxicity were significantly higher for drug loaded nanoparticles than free drug. These results suggested porous inorganic nanoparticles might be a promising intracellular carrier for hydrophobic anticancer drugs.

OBJECTIVETo compare the regulatory effects of electro-acupuncture (EA) at acupoints Zusanli (ST36) and Hegu (LI4) on the visceral hyper-sensitivity in the rat model of irritable bowel syndrome (IBS), and to explore the acting targets and specialty of acupoints.

METHODSExcept 8 rats of the normal control group, the rest 32 rats were prepared to set up the IBS models. IBS animal model was prepared by enema with acetic acid. Model rats were divided into three groups. Except for rats in the model group for control, those in the other two groups were treated 20 min by EA on ST36 (EA-ST36) and LI4 (EA-LI4) respectively for 2 weeks to observe the effect on behavior response of viscera sensitivity. The changes of neuropeptide (NPY), the somatostatin (SS) levels in blood and tissues of brain and intestine were monitored as well.

RESULTSThe volume thresholds for abdomen uplifting and back hunching were obviously increased after EA-ST36 (P<0.05), but showed insignificant change after EA-LI4. NPY contents lowered and SS contents increased in model rats; both EA-ST36 and EA-LI4 could raise the level of thalamic NPY (P<0.01 and P<0.05, respectively), but showed insignificant effects on NPY in colonic tissue. As for SS content, its colonic level could be reduced by EA-S36 and EA-LI4 (P<0.01 and P<0.05, respectively), however, its blood level was affected only by EA-ST36 (P<0.05).

CONCLUSIONSEA-ST36 or EA-LI4 could regulate the NPY in thalamus and SS in colonic tissue, the former could affect blood level of SS as well. It is deemed that NPY and SS may be the key substances for regulating the action of acupuncture in the brain-intestinal axis; their different levels could be regarded as an indicator for the functional difference between the acupoints.

Acupuncture Points ; Animals ; Brain ; metabolism ; Electroacupuncture ; methods ; Irritable Bowel Syndrome ; metabolism ; physiopathology ; Neuropeptide Y ; metabolism ; Rats ; Rats, Wistar ; Somatostatin ; metabolism ; Viscera ; physiopathology