Objective: Analytic hierarchy process (AHP) was used to establish a comprehensive evaluation method of multiple indexes of traditional Chinese medicine, and the best processing technology of Sarcandra glabra was optimized. Methods: On the basis of single factor experiment, the comprehensive score of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraxidin, astilbin and rosmarinic acid content was used as evaluation index by AHP, and drying temperature and drying time were used as influencing factors. The purification process of S. glabra leaves was determined by single factor investigation. Taking soaking time, soaking time, cutting length and drying temperature as influencing factors, the optimal cutting process of S. glabra stems was optimized by central composite design-response surface method. Multi-index analytic hierarchy process (AHP) was used to analyze and verify the optimal process. Results: The best processing technology of S. glabra stems was as follows: stem Rob water washed once, soaked in 12 times of water for 1 h, moistened and softened for 3 h, cut into 15 mm segments after taking out, and baked at 50 ℃ for 2 h; The processing method of leaves was as follows: leaf Rob water washed once and dried at 50 ℃ for 4 h; Then mixed the processed stems and leaves of S. glabra at 2:1. The deviation between the verification value and the prediction value of the comprehensive score of the optimal S. glabra stem by softening and cutting method was 4.70%. Conclusion: The processing technology is simple, stable and feasible. The obtained parameters can provide scientific basis for the research and production of S. glabra, and AHP can provide reference for the multi-index evaluation of traditional Chinese medicine.

We designed specific primers and fluorescence-labeled probes to develop real-time and conventional RT-PCR assays for detection of human coronavirus NL63 or HKU1. Subsequently, experiments were undertaken to assess diagnostic criteria such as specificity, sensitivity and reproducibility. The detection limit of the real-time RT-PCR assays was 10 RNA copies per reaction mixture. No cross-reactivity was observed between RNA samples derived from designed HCoV and other HCoV or human metapneumovirus. A total of 158 nasopharyngeal swab specimens collected from adult patients with acute respiratory tract infection in Beijing were screened for the presence of human coronavirus NL63 and HKU1 by using real-time RT-PCR and conventional RT-PCR method. The fluorescence quantitative RT-PCR method detected six specimens positive for human coronavirus NL63, five specimens positive for human coronavirus HKU1; and conventional RT-PCR method detected three HCoV-NL63 positive and three HCoV-HKU1 positive, respectively. The convention RT-PCR products of positive samples were obtained and sequence analysis confirmed the reliability of the above methods. In summary, the real-time RT-PCR assay for HCoV- NL63 or HKU1 was more sensitive than conventional RT-PCR and with less time (less than 4 hours) for completion. It may be suitable for molecular epidemiological surveillance and clinical diagnosis for human coronavirus NL63 and HKU1.
Coronavirus ; classification ; genetics ; isolation & purification ; Humans ; Nasopharynx ; virology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To explore the effect of dance intervention on the cognitive function and balance function in the elderly with mild cognitive impairment and its possible mechanism through systematic review of related literatures. Methods The literatures of randomized controlled trials on the impact of dance intervention on the cognitive function and balance function of the elderly with mild cognitive impairment were searched in the Cochrane library, PubMed, CBM, Web of Science, EMBASE, CNKI, VIP, Wanfang data and other databases, from establishment to June 7th, 2021. Two researchers screened the literature according to the inclusion and exclusion criteria, extracted data, and evaluated the quality. Results The scores of Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were better in the dance intervention experimental group than in the control group, as well as the scores of Wechsler Memory Scale (WMS), Trail Making Test (TMT)-B and Verbal Fluency Test. However, no significant difference was found in the response time in TMT-A between two groups. Dance intervention could improve the scores of Berg Balance Scale. Conclusion Dance intervention can improve overall cognitive function, memory function, executive function and balance function of the elderly with mild cognitive impairment.

The delay of diabetic foot not only brings painful experience to patients, but also increases the care time and caregiver burden, which seriously affects quality of life of patients and caregivers. This article summarized the definition, research tools, current situation, influencing factors, positive feelings of caregivers and intervention measures of caregivers for diabetic foot patients, aiming to provide a theoretical basis for adoption of targeted intervention measures.

BACKGROUND: With the ongoing worldwide coronavirus disease 2019 (COVID-19) pandemic, an increasing number of viral variants are being identified, which poses a challenge for nucleic acid-based diagnostic tests. Rapid tests, such as real-time reverse transcription-polymerase chain reaction (rRT-PCR), play an important role in monitoring COVID-19 infection and controlling its spread. However, the changes in the genotypes of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants may result in decreased sensitivity of the rRT-PCR assay and it is necessary to monitor the mutations in primers and probes of SARS-CoV-2 detection over time. METHODS: We developed two rRT-PCR assays to detect the RNA-dependent RNA polymerase (RdRp) and nucleocapsid (N) genes of SARS-CoV-2. We evaluated these assays together with our previously published assays targeting the ORF1ab and N genes for the detection and confirmation of SARS-CoV-2 and its variants of concern (VOCs). In addition, we also developed two rRT-PCR assays (S484K and S501Y) targeting the spike gene, which when combined with the open reading frames (ORF)1ab assay, respectively, to form duplex rRT-PCR assays, were able to detect SARS-CoV-2 VOCs (lineages B.1.351 and B.1.1.7). RESULTS: Using a SARS-CoV-2 stock with predetermined genomic copies as a standard, the detection limit of both assays targeting RdRp and N was five copies/reaction. Furthermore, no cross-reactions with six others human CoVs (229E, OC43, NL63, HKU1, severe acute respiratory syndrome coronavirus and Middle East respiratory syndrome coronavirus) were observed using these assays. In addition, the S484K and S501Y assays were combined with the ORF1ab assay, respectively. CONCLUSIONS Four rRT-PCR assays (RdRp, N, S484K, and S501Y) were used to detect SARS-CoV-2 variants, and these assays were shown to be effective in screening for multiple virus strains.
COVID-19 ; Humans ; RNA, Viral/genetics* ; Real-Time Polymerase Chain Reaction ; Reverse Transcription ; SARS-CoV-2 ; Sensitivity and Specificity

Objective To express the nuclear capsid protein (N protein) and the spike protein (S protein) of HCoV-HKU1, and to develop the corresponding serum assay for antibody detection. Methods The N protein of HCoV-HKU1 was expressed in E. Coli, anti-N antibody assay was established using Western Blotting with turn-based membrane. HCoV-HKU1 S protein was constructed in the eukaryotic expression plasmids, and confirmed by Western Blotting, S antibody assay was established using indirect immunofluorescence assay (IFA). We analyzed anti-S and anti-N antibody among 100 normal adult serum.Results Expression of S and N protein were confirmed;100 normal adult serum were analyzed using the established serological detection assay, in which HCoV-HKU1 S antibody positive rate was 47% , N antibody positive rate was 48% , Both S and N antibodies positive were 21% , Both S and N antibodies negative were 22%. Co-detection S and N antibody was achieved 74% positive rate. Conclusion The methods we established here could be used for serological analysis of HCoV-HKU1. Either detection of HCoV-HKU1 S or N antibodies achieved good results. Higher positive detection rate of anti-S or anti-N antibody was found in the normal adults.

OBJECTIVETo study the human immunodeficiency virus (HIV) status through heterosexual transmission in Yining city and to provide information on effective intervention measures.

METHODSCohort of HIV sero-discordant couples identified from 1997 to 2000 was formed. Proportional risk model was used to analyze the time of HIV sero-conversion and the related factors. All the recruiters were under informed consent.

RESULTSThrough following on 22 sero-discordant couples, we found that the incidence density (ID) of HIV sero-conversion was 32.49/100 person-year (PY) with 33.74/100 PY for women. In the proportional hazard model, the course of sero-conversion was only 2.43 years and the frequency of sexual contact was statistically significant (>or= 3 times/week vs. < 3 time/week: RR = 1.984, 95% CI: 1.045 - 3.767), indicating this factor was related to the hazard of HIV sero-conversion. However, the viral load of HIV infections has no such effect on HIV sero-conversion of their spouses. In addition, the ratio of CD4(+)/CD8(+) was lower in spouses of HIV sero-conversion than that in spouses of HIV non-sero conversion (t test: t = 4.77, P < 0.01).

CONCLUSIONIn order to control HIV transmission among general population, we suggested that HIV/AIDS counseling and testing be developed for pre-marital people in the region with high HIV prevalence.

Blotting, Western ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV ; immunology ; HIV Infections ; epidemiology ; immunology ; Humans ; Incidence ; Male ; Proportional Hazards Models ; Sexual Behavior ; Sexually Transmitted Diseases, Viral ; epidemiology ; immunology

The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.
Base Sequence ; Blotting, Western ; Coronavirus NL63, Human ; chemistry ; Fluorescent Antibody Technique, Indirect ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Plasmids ; Recombinant Proteins ; biosynthesis ; Spike Glycoprotein, Coronavirus ; Vaccinia virus ; genetics ; Viral Envelope Proteins ; biosynthesis ; genetics

To investigate the genetic stability (including the vector of vaccinia virus and six foreign genes: gp160, gag, pol, rev, tat and nef) of the HIV-1 non-replicating recombinant vaccinia virus (rNTV-C). rNTV-C was serially passaged to passage 25 (P25) in primary chicken embryo fibroblast (CEF). P9, P12, P15 and P25 were selected to study the genetic stability in four aspects, including the genetic stability of viral vector, the genetic stability of six foreign genes, the expressing stability of foreign genes and the genetic loss of foreign genes. The results showed that the viral vector was non-replicated vaccinia virus of Tiantan strain and was passaged stably; foreign gene sequences matched with designed sequences, the insert sites were right, and the nucleotide mutation rate was less than one over ten thousands within different passages of rNTV-C; the target proteins could be expressed effectively, and the expression level was stable within different passages of rNTV-C; the genetic loss of gag and nef was less than 5% within different passages of rNTV-C. The above results provided important data for the vaccine production.
Animals ; DNA, Recombinant ; genetics ; Fibroblasts ; metabolism ; virology ; Gene Expression ; Genes, Viral ; genetics ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; HIV-1 ; genetics ; Sequence Analysis, DNA ; Vaccinia virus ; genetics

The purpose is to screen and identify the specific H-2d restricted T-cell epitopes. These epitopes are used to investigate the cellular immune response of BALB/c (H-2d) mice immunized with a HIV-1 vaccine which expresses six antigens of gp160, gag, pol, rev, tat and nef of HIV subtype B'/C. A replicating DNA vaccine and a non-replicating recombinant vaccinia virus vector, both expressing the six antigens mentioned above, were used to immune BALB/c (H-2d) mice in a prime-boost regiment. The six peptide libraries of HIV B'/C corresponding respectively to the six complete antigens were pooled according to a designed matrix format and used to test for IFN-gamma production from splenocytes of immunized mice by an enzyme-linked immunospot (IFN-gamma ELISPOT) assay. The ELISPOT data indicated that two of seven Gag-specific T-cell epitope peptides were identified to be the novel epitopes. One of three Pol-specific T-cell epitope is unreported. One novel epitope was confirmed in two gp160-specific T-cell epitope peptides. One Nef-specific T-cell epitope was identified. Three Tat-specific T-cell epitope peptides were continuous sequences in Tat peptide library and all contained either complete or partial sequence reported. Rev-specific T-cell epitope was not be found. The specific T-cell epitopes (H-2d restricted) were identified by IFN-7 ELISPOT assay, which could be used to detect the cellular immune response of BALB/c mice immunized with the HIV-1 vaccine expressing six antigens of gp160, gag, pol, rev, tat and nef of HIV subtype B'/C.
Amino Acid Sequence ; Animals ; Enzyme-Linked Immunospot Assay ; methods ; Epitopes, T-Lymphocyte ; chemistry ; genetics ; immunology ; Female ; H-2 Antigens ; chemistry ; genetics ; immunology ; HIV Antigens ; chemistry ; genetics ; immunology ; HIV Infections ; immunology ; virology ; HIV-1 ; classification ; genetics ; immunology ; Histocompatibility Antigen H-2D ; Humans ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Peptide Mapping ; methods