Biodiesel, a nontoxic,cleaning, renewable and biodegradable fuel, is expected as a substitute for conventional fossil diesel. There are three main approaches to produce biodiesel, alkali-catalysis processing, enzymatic-catalysis processing and supercritical processing. With the unique property of energy-saving and environment-friendly, enzymatic-catalysis appears a great potential for industrial application. The main bottleneck of this technology is high cost and low stability of the lipase, as well as the inactivation of lipase by methanol and so on. To settle the problem, several methods have been used including the fixed-bed bioreactor, enzyme immobilized processing, whole-cell biocatalyst, changing addition method of methanol, developing of novel acyl acceptor, enhancing methanol resistance of lipase. The main problems and the relative strategy research of the enzymatic-catalysis technology were sum up.

Lipase gene from Penicillium expansum(lip07) was cloned and over-expressed in Pichia pastoris.a random mutant named ep8,which contained a single amino acid substitution,was obtained by using the lip07 as an error-prone PCR template in previous study.ep8 shows higher thermostability than that of lip07,To further improve the thermostability of the lipase,the Lys of wild-type(lip07) and mutant(ep8) in 202 were substituted by Ala using the Overlap extension PCR technique respectively.The mutant genes(lip07-K202A and ep8-K202A) were subcloned into pAO815,and then transformed into the Pichia pastoris GS115 for extracelluar expression,respectively.15% SDS-PAGE analysis indicated that the molecular mass of PEL-ep8-K202A and PEL-lip07-K202A are both about 28kDa,which is same with the wild-type lipase.The Tm of PEL-ep8-K202A is 41.66℃,2.63℃ higher than that of the wild-type(39.03℃) and 1.21℃ higher than the random mutant(PEL-ep8:40.45℃);the Tm of single mutant(PEL-lip07-K202A) is 37.08℃,2℃ lower than that of the wild-type lipase.

Adult ; Biopsy, Needle ; Diagnosis, Differential ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; therapy ; Male ; Nephritis, Interstitial ; pathology ; Renal Dialysis ; Sarcoidosis ; pathology ; therapy ; Tuberculosis, Renal ; pathology

AIM: To investigate the antitumor activity and mechanism of Paecilomyces tenuipes polysaccharide(PTPS).METHODS: PTPS-I was obtained by water extraction and alcohol precipitation,and purified by DEAE-cellulose and Sephadex G-100 chromatography.Human erythroleukemia cell line K562,laryngocarcinoma cell line Hep2 and hepatic carcinoma cell line SMMC-7721were co-cultured with PTPS-I or the conditioned medium which prepared with PTPS-I-stimulated human mononuclear cells(PTPS-I-MNC-CM),and the proliferation of tumor cells was determined.The cell counting kit-8(CCK-8) was used to determine the proliferation of MNCs.The FQ-RT-PCR was applied to investigate the expression of TNF-? and IL-6 mRNA in MNCs.RESULTS: PTPS-I-MNC-CM inhibited the proliferation of K562,Hep2 and SMMC-7721 cells in vitro(P

Objective To explore a way to induce sputum from the deep tracheal and to analyze the secretion. Methods A total of 80 patients were randomly divided into two groups: the control group received 3% hypertonic saline aerosolized, while the experiment group relieved 3% hypertonic saline, combined with Ambroxol Hydrochloride. Results The experiment group induced more sputum and had high success rate. Difference between the two groups was significant (P <0. 01). Conclusions It is satisfied to select the method of % hypertonic saline, combined with Ambroxol Hydrochloride. for inducing sputum among patients without sputum or viscous sputum

OBJECTIVETo detect the expression of human telomerase reverse transcriptase (hTERT) protein and mRNA in bile duct carcinomas and the adjacent tissues and to elucidate its role in bile duct carcinogenesis.

METHODSThe expression of hTERT protein and hTERT mRNA in the formalin-fixed paraffin-embedded specimens of 71 cases of bile duct cancers and 39 cases of adjacent tissues was detected by streptavidin-peroxidase immunostaining and in situ hybridization. The correlation was analysed statistically between the expression of hTERT protein and mRNA and clinicopathological parameters bile duct carcinomas.

RESULTSThe positive rate of hTERT protein expression and mRNA expression in malignant specimens was 78.9% (56/71) and 67.6% (48/71), while that in the adjacent tissues was 35.9% (14/39) and 23.1% (9/39), respectively. All the positive signals were found in the hyperplastic biliary epithelia. No significant correlation was established between hTERT expression and clinicopathological parameters.

CONCLUSIONhTERT gene transcription and protein expression is most likely involved in the proliferation and malignant transformation of bile epithelia and the malignant progression of bile duct carcinomas. The detection of hTERT expression may serve elucidating the carcinogenesis of bile duct.

Adult ; Aged ; Aged, 80 and over ; Bile Duct Neoplasms ; enzymology ; pathology ; DNA-Binding Proteins ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; RNA, Messenger ; analysis ; Telomerase ; analysis ; genetics

Optimization of sensor array is a significant topic in the application of electronic nose (EN). Stepwise discriminant analysis and cluster analysis combining with screening of typical index were employed to optimize the original array in the classification of 100 samples from 10 kinds of traditional Chinese medicine based on alpha-FOX3000 EN. And the identification ability was evaluated by three algorithm including principle component analysis, Fisher discriminant analysis and random forest. The results showed that the identification ability of EN was improved since not only the effective information was maintained but also the redundant one was eliminated by the optimized array. The optimized method was eventually established, it was accurate and efficient. And the optimized array was built up, that is, S1, S2, S5, S6, S8, S12.
Algorithms ; Biosensing Techniques ; methods ; Cluster Analysis ; Discriminant Analysis ; Drugs, Chinese Herbal ; classification ; isolation & purification ; Electronic Nose ; Medicine, Chinese Traditional ; Principal Component Analysis ; Reproducibility of Results ; Smell

AIMTo investigate the glycosidic constituents in the rhizomes of Alpinia officinarum Hance.

METHODSThe isolation and purification of glycosides were done with column chromatography on macro porous resin, polyamides and Sephadex LH-20, whilst the structure elucidation was done by HRCI-MS and NMR (1D and 2D) methods.

RESULTSA glycosidic ester identified as 4'-hydroxy-2'-methoxyphenol-beta-D-{6-0-[4"-hydroxy-3", 5"-dimethoxy (benzoate)]}-glucopyranoside (I), along with a known compound n-butyl-beta-D-fructopyranoside (II), were isolated and characterized.

CONCLUSIONI was found to be a new compound, named as alpinoside A, whilst II was isolated from the genus Alpinia for the first time.

Alpinia ; chemistry ; Fructose ; analogs & derivatives ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

AIMTo develop a capillary gas chromatographic method for the determination and pharmacokinetic study of patchouli alcohol in rat plasma after iv administration.

METHODSThe drug was extracted with ethyl acetate. Eugenol was used as internal standard. The separation was carried out on a HP-5MS quartz capillary column, with high-purity nitrogen as carrier gas and flame ionization detector (FID) as detector. The column temperature was maintained at 80 degrees C for 1 min and then programmed to 200 degrees C at a rate of 15 degrees C x min(-1); it was held at 200 degrees C for 1 min, and then programmed to 290 degrees C at a rate of 60 degrees C x min(-1); the final temperature was held for 1 min. The temperature of both injector and detector was set at 290 degrees C.

RESULTSThe standard curve was linear from 25 to 5 000 microg x L(-1) in rat plasma. The recovery of this method was from 90.0% to 110.0% with satisfactory relative standard deviation (RSD) less than 10.0%. The pharmacokinetic parameters demonstrated patchouli alcohol were consistent with the two-compartment open model and showed linear pharmacokinetics. The T1/2beta, AUC and MRT of patchouli alcohol in patchouli oil were all higher than that of patchouli alcohol.

CONCLUSIONThis method is quick, precise and reliable. The pharmacokinetics of patchouli alcohol is different from that of patchouli alcohol in patchouli oil.

Animals ; Area Under Curve ; Injections, Intravenous ; Lamiaceae ; chemistry ; Male ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sesquiterpenes ; blood ; isolation & purification ; pharmacokinetics

Abstract:Objective　To investigate the clinical characteristics and early diagnostic methods of patients with Talaromyces marneffei infection, so as to reduce the mortality of patients. Methods The clinical characteristics and microbiological analysis data including fungal culture, smear examination and mass spectrometry were collected from 18 patients with Talaromyces marneffei infection in the Department of Respiratory Medicine, Department of Tuberculosis, and Department of Critical Respiratory Medicine in Fuzhou Pulmonary Hospital from January 2017 to December 2021, and descriptive analysis was conducted. Results　All the 18 patients were confirmed to be infected with Talaromyces marneffei by conventional culture and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS). The main infection sites of 18 patients with Talaromyces marneffei infection were lungs and lymph nodes, and the patients were accompanied by clinical manifestations such as cough, sputum and fever. The imaging features such as patchy shadows, mediastinal lymph node shadows and nodular shadows were common. Microbiological testing showed a statistically significant difference between smear and culture with a higher positive culture rate (χ2=13.74, P<0.05). The positive rate of blood culture in microbiological test was 60.0% (9/15), the positive rate of bronchial lavage fluid culture was 26.7% (4/15), the positive rate of sputum culture was 5.6% (1/18), one case each of pus, bone marrow, pleural fluid and cerebrospinal fluid was positive for culture and the other cases were negative, one case of sputum and one case of pus were positive for smear and the rest were negative. Colony characteristics showed that the colony morphology was mycelial phase at 25 ℃, producing red pigment, and the branching pattern of the penicillus was seen microscopically as monoverticillate or biverticillate; At 35 ℃, the yeast phase appeared at the initial stage, and then the mycelium phase changed after 5-6 days; the yeast phase was observed at 37 ℃, and yeast-like cells were seen under the microscope. All 18 patients with Talaromyces marneffei infection got better after using antifungal drugs. Compared with non-HIV patients with Talaromyces marneffei infection, leukopenia and anemia were common in HIV patients with Talaromyces marneffei infection, and the differences were statistically significant (P<0.05). 　Conclusions The infection of Talaromyces marneffei can be divided into localized type and disseminated type, which usually invade the lungs, skin, lymph nodes and other places. The main manifestations of patients are fever, cough, phlegm and other atypical symptoms. At present, the diagnosis of Talaromyces marneffei infection is mostly based on the fungal culture test, and the application of MALDI-TOF MS method can effectively shorten the diagnosis time of Talaromycosis marneffei. Clinical characteristics combined with microbiological analysis provide an objective basis for early diagnosis of patients with Talaromyces marneffei infection, and timely use of antifungal therapy can improve the prognosis of patients.