The way of "extracting-salting-chromatography" was used to purify the phycoerythrin and phycocyanin from Porphyra yezoensis in process scale-up.First,by comprehensive comparison of efficiency,the Sephadex G-25 was selected from four resins (Sephadex G-25、G-100、S-300 and CL-6B) as the best choice used in crude extract desalting of phycobiliprotein.Then the preparation process of phycobiliprotein was scaled-up with raw material(Porphyra yezoensis) increased from 1g to 20g,and finally to 400g.The results indicated that the yields of purified phycoerythrin and phycocyanin (absorption spectra purity above 3.2) increased during according to process scale-up,with 0.323% phycoerythrin and 0.148% phycocyanin obtained from 400g frozen Porphyra yezoensis blades respectively.It is no doubt that the process involved in the experiment is a potential way for large scale preparation of phycobiliproteins of high purity.

Hepatitis E, an acute infectious disease transmitted via the fecal-oral route, is caused by hepatitis E virus. However, no effective treatment currently exists for hepatitis E, and the only epidemic control approach is vaccination. But so for there are no commercial vaccine for hepatitis E available in the world. To find a new expression system to develop recombinant hepatitis E vaccine, in this study the expression system of methylotrophic yeast Hansenula polymorpha was used to express the gene encoding amino acid 112 - 607 of the open reading frame 2 (ORF2) of hepatitis E virus (HEV) genotype IV. In order to achieve high expression level, the coding sequence was optimized according to codon usage bias of Hansenula polymorpha and synthesized through overlapping PCR. Subsequently the gene was subcloned into the multi-copy expression vectors of Hansenula polymorpha, which include pDGXHP1.0 (MOX promotor), pDGXHP2.0 (MOX promotor) and pDGXHP2.1 ( FMD promotor). The series of one-copy and multi-copy recombinant plasmids were transformed into ATCC26012(Ura3-) by electroporation. The transformants were cultured in selection media MDL and screened for the existence of foreign gene by PCR. Then the strains were induced in MM media and the expression products were detected by SDS-PAGE, ELISA and Western blot assays to select the high-level expression strains. The result of SDS-PAGE showed that the HEV ORF2 expression product was accumulated up to 12% of total cellular protein and its molecular weight is 56kD. The expression product showed high immunoreactivity detected by ELISA and the highest titer is 1:2048. The result of Western blot demonstrated that the expression product could be specifically recognized by the polyclonal antibody against HEV. The successful expression of HEV ORF2 protein in Hansenula polymorpha provides foundation for the further development of recombinant subunit vaccine against hepatitis E.
Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Gene Expression Regulation, Viral ; Genotype ; Hepatitis E ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; metabolism ; Humans ; Pichia ; genetics ; Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; metabolism ; Viral Hepatitis Vaccines ; immunology ; Viral Proteins ; genetics ; immunology ; metabolism

OBJECTIVETo study how to perform the hearing screening on the infants in the rural area.

METHODSThree thousand nine hundreds and twenty-two infants, about 84% of them from rural, were born in the People Hospital of LaiZhou City from January to December in 2004. The infants were performed fast hearing screening by transient evoked otoacoustic emission (TEOAE) after the birth in 2-7 days. The fail cases were checked again after 4-6 weeks, and then were diagnosed if they still failed after following-up.

RESULTSThe infants (3612/3922, 92.1%) have been checked by TEOAE, and the examination was free in the poverty cases. The rate passed on the first check was 69.96% (2527/3922), but 1085 infants failed (30.4%), while 310 infants have not been checked (7.9%). In the 1085 cases that should be rechecked, there was only 633 cases (58.34%) accepted the check on time, while 452 cases (41.66%) missed. In the 163 cases with high-risk infants in 2004, 114 infants (69.96%) were checked, but 49 infants (29.04%) were not checked. Fourteen cases failed in the recheck, and 11 of them were checked by ABR. Two cases were found to be moderate and severe hearing loss in binaural respectively and 4 cases with mild hearing loss in monaural while 3 cases were normal.

CONCLUSIONSIt is necessary and viable for the infants on hearing screening in the rural area It should be set up and perfected the model for infants on hearing screening in rural area as soon as possible; it should be free for the poor infants to make sure everyone enjoy the health care.

China ; Hearing Tests ; Humans ; Infant, Newborn ; Neonatal Screening ; methods ; Otoacoustic Emissions, Spontaneous ; Rural Population

Objective: Coronavirus Disease 2019 (COVID-19) has been diagnosed as an epidemic disease characterized by cold and dampness pathogens in TCM clinic. Due to many Chinese medicines with different functions were used in the treatment of COVID-19, it is very important to find the law of application of damp-removing traditional Chinese medicine with high frequency application, with view to providing a reference for the use and research of Chinese medicine to further control the pandemic. Methods: The publicly released diagnosis and treatment programs issued by the National Health Commission and Health Commission of provinces, autonomous regions and municipalities, and Chinese herbs prescription information in these were collected, a database was established, and Excel and Graphpad 8.0 software were used to analyze the frequency of use of various Chinese medicines, the frequency and property characters including five flavors (bitter, pungent, sweet, sour, and salty) and four natures (warm, hot, cool, and cold) and channel tropisms of Chinese medicine for removing dampness. Results: A total of 137 prescriptions of Chinese medicine for treating COVID-19 were collected, including 178 TCMs showing functions of resolving phlegm, relieving cough and asthma, resolving dampness, clearing damp and inducing dieresis, clearing heat, tonifying deficiency, and relieving exterior syndrome, in which the TCMs with the first four functions that we called the dampness-removing TCMs, accounted for 35.78%. Also a number of TCMs in the rest functions showed removing-dampness. The first four functions were divided into subfunctions including aromatic resolving dampness, clearing heat and drying dampness, drying dampness and tonifying spleen qi, drying dampness and removing phlegm, inducing diuresis and relieving swelling, inducing diuresis and relieving exterior syndrome. Among them, the most frequently used TCMs was Ephedrae Herba, followed by Citri Reticulatae Pericarpium, Pogostemonis Herba, Pinelliae Rhizoma, Poria, Scutellariae Radix, and Atractylodis Rhizoma. The property character analysis in the dampness-removing TCMs showed that bitter and pungent were largely present and sour and astringent were absent, warm and hot were dominant; And the lung, spleen, stomach, large intestine, bladder were main channel tropisms. Conclusion: Dampness-removing TCMs are the first important type of traditional Chinese medicine to be considered in the treatment of COVID-19 in Chinese medicine. The application of dampness-removing TCMs in the treatment of COVID-19 needs to be combined with its application law. This study may provide meaningful and useful information on further research to investigate the effective compounds from the dampness-removing Chinese medicine with high frequency application, and also provide a reference for the clinical treatment of COVID-19 accurately against dampness evil with dampness-removing traditional Chinese medicines.

OBJECTIVE: Here, we explored molecular changes that could potentially mediate healing effects of Gua Sha - a method employed by the Chinese traditional medicine with proven track records of safe and efficient applications dating back to ancient times as well as support from randomized controlled trials performed by modern medical studies - yet remaining almost entirely unexplored by the modern-day high-throughput methods of the -omics sciences. METHODS: We investigated transcriptome changes occurring shortly after Gua Sha treatment in the whole blood of healthy volunteers using bulk RNA-seq analysis. We applied various analytical tools to identify genes with consistent expression changes in multiple individuals in response to Gua Sha and their networks. RESULTS: We found that while the changes were very subtle and individual-specific, we could identify consistent upregulation of three histone genes. Further analysis of the potential regulatory networks of these histone genes revealed the enrichment of functions involved in the immune response and inflammation. CONCLUSION The significance of these results in the context of potential effects of Gua Sha and the next steps in exploring the molecular mechanisms of action of this technique are discussed.
Humans ; Medicine, Chinese Traditional/methods* ; Histones ; Gene Expression

Cinnamomi Ramulus is a dry tender branch of Cinnamomum cassia Presl, which is a multifunctional traditional Chinese medicine(TCM). Cinnamomi Ramulus has different efficacy under different compatibility environment. Complet medicine is a commonly used and relatively fixed compatibility form of two drugs, and it is the smallest unit in the compatibility of TCM. The four kinds of complet medicine commonly used in Cinnamomi Ramulus were as follows: the diaphoretics pungent in flavour and warm in property pairs include Cinnamomi Ramulus Ephedrae Herba and Cinnamomi Ramulus Bupleuri Radix complet medicine. The stasis dredge collaterals pairs involve Cinnamomi Ramulus Poria and Cinnamomi Ramulus Persicae Semen complet medicine. The regulation Qi and Blood Cinnamomi Ramulus Paeoniae Radix Alba complet medicine. Wenda Tongyang pairs consist of Cinnamomi Ramulus Aconiti Lateralis Radix Praeparaia, Cinnamomi Ramulus Astragali Radix, and Cinnamomi Ramulus Glycyrrhizae Radix et Rhizoma complet medicine. After compatibility, some changes have taken place in the chemical composition of complet medicines. For example, after compatibility of Cinnamomi Ramulus and Ephedrae Herba, the content of effective components of both herbs decreases, and the chemical constituents that are not found in single herbs are produced. After compatibility of Cinnamomi Ramulus and Bupleuri Radix, the dissolution of active ingredients is related to compatibility ratio. The active ingredients of Cinnamomi Ramulus Poria, Cinnamomi Ramulus Paeoniae Radix Alba pair and Cinnamomi Ramulus Aconiti Lateralis Radix Praeparaia have also changed to some extent.The content of active ingredients in Astragali Radix and Glycyrrhizae Radix et Rhizoma both increase after compatibility with Cinnamomi Ramulus. Different complet medicine have different pharmacological effects, Cinnamomi Ramulus Ephedrae Herba complet medicine have the effect of transpiration and antipyretic, Cinnamomi Ramulus Bupleuri Radix complet medicine can analgesia. Cinnamomi Ramulus Poria complet medicine have the effect of diuretic, improve myocardial ischemia and so on. Cinnamomi Ramulus Persicae Semen complet medicine have the anti-coagulation action.Cinnamomi Ramulus Paeoniae Radix Alba complet medicine have the anti-inflammatory and analgesic activities. Cinnamomi Ramulus Aconiti Lateralis Radix Praeparaia complet medicine have the effect of dispelling cold to relieve pain. Cinnamomi Ramulus Astragali Radix complet medicine have the many effects in relieving anti-coagulation, antioxidation and anti-myocardial ischemia. Cinnamomi Ramulus Glycyrrhizae Radix et Rhizoma complet medicine displays diverse activities, including antiarrhythmic, antithrombosis. In this paper, the chemical constituents and pharmacology of four kinds of complet medicine of Cinnamomi Ramulus were reviewed, which provided a reference for the better open utilization of Cinnamomi Ramulus complet medicine.