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

OBJECTIVETo know the etiology, prevalence, clinical symptoms associated with the infection of the HCoV-229E in the respiratory specimens sampled from adult patients in Beijing.

METHODS158 nasopharyngeal swab specimens were collected from adult patients with fever in Beijing between October and December, 2007. We performed the screening of HCoV-229E by real-time RT-PCR and sequencing of HCoV-229E gene fragments derived from conventional PCR. At meantime, we also screened the HCoV-229E positive samples for the co-infection with HCoV-NL63, HCoV-HKU1 and HMPV by real-time RT-PCR. Finally, demographic and clinical data associated with HCoV-229E infection were examined retrospectively.

RESULTSWe detected 103 (62.5%) of 158 specimens were positive for HCoV-229E by real-time RT-PCR. When tested for other respiratory viruses, 26 HCoV-229E positive patients were found to be co-infected with other viruses. Of which HCoV-NL63 was observed in 3 specimens (11.5%), HCoV-HKU1 in 3 (11.5%) and HMPV in 20 (76.9%). The main clinical manifestations were noted as: headache (in 70.9%), sore throat (69%), chills (68%), cough (33%), sputum (21.3%), rhinorrhea (21.4%), nasal obstruction (16.5%), and a few of patients were visible as vomiting (6.8%), dyspnea (3.9%), diarrhea (in 1.9%). The rate of HCoV-229E infection in adult patients was found no relative with age and gender.

CONCLUSIONOur data showed that HCoV-229E is a common and important pathogen in adult patients with acute respiratory symptoms but usually resulted in milder influenza-like illnesses. There might have a local outbreak of HCoV-229E infection in Beijing, Oct-Dec, 2007.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Coronavirus 229E, Human ; classification ; genetics ; isolation & purification ; Coronavirus Infections ; diagnosis ; epidemiology ; virology ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Respiratory Tract Infections ; diagnosis ; epidemiology ; virology ; Retrospective Studies ; Young Adult

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.

OBJECTIVETo 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.

METHODSThe 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.

RESULTSExpression 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.

CONCLUSIONThe 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.

Antibodies, Viral ; blood ; immunology ; Capsid Proteins ; genetics ; immunology ; Cell Line ; Coronavirus ; genetics ; immunology ; isolation & purification ; physiology ; Coronavirus Infections ; blood ; diagnosis ; immunology ; virology ; Humans ; Membrane Glycoproteins ; genetics ; immunology ; Serologic Tests ; methods ; Spike Glycoprotein, Coronavirus ; Viral Envelope Proteins ; genetics ; immunology

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

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

OBJECTIVETo develop and optimize the molecular detection assays for recently identified human coronavirus (HCoV) infection.

METHODSBased on the 208 base pair(bp) sequence of novel HCoV reported by HPA of UK, we designed and obtained several pairs of primer (F-1, R-1; F-2, R-2) and Taqman probes (TZ1,TZ2) for detection of novel HCoV. Two of probes were modified with LNA (LNA-TZ1, LNA-TZ2). Then, RT-PCR and various real time RT-PCR assays were developed and optimized in this study. We also compared our assays with the real time RT-PCR assays reported recently by Europe team based on upE or ORF1b target.

RESULTSThe RT-PCR or real time RT-PCR assays for novel HCoV were developed without cross-reactivity with other HCoV and several common respiratory viruses using clinical specimen panel. The analytical sensitivity of assays were less than 50-500 copies per reaction and the detection was improved when Taqman probe modified with LNA-tagged, compared to no LNA-tagged in real time RT-PCR assays. The upE and LNA-TZ1 based assays were better than others.

CONCLUSIONThe molecular detection sensitivity and specificity of TaqMan-based real time PCR assay could be improved when probe tagged with LNA. The upE or LNA-TZ1 based real time RT-PCR assay was recommend for detection of novel HCoV. This study laid a foundation for improving the performance of novel HCoV detection.

Coronavirus ; classification ; genetics ; isolation & purification ; DNA Primers ; genetics ; Humans ; Oligonucleotides ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

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

OBJECTIVETo describe the characteristics and factors associated with long-term retention for methadone maintenance treatment (MMT) patients.

METHODSThis study was conducted in eight MMT clinics located in Sichuan, Yunnan, Guangxi, Guizhou and Zhejiang provinces. Five hundred and thirty-nine MMT patients who enrolled in MMT clinics in 2004 and retained in treatment by June 2010 were selected as study subjects. Chi-square tests were used to compare the demographics and drug abuse history at enrollment and treatment characteristics during the follow-up period between continuous treatment patients and discontinuous treatment patients.

RESULTSOf the 539 patients, 110 (20.4%) were continuous treatment patients whereas 429 (79.6%) were discontinuous treatment patients. Of these 429 discontinuous treatment patients, 84.1% (361/429) had 2-4 treatment episodes whereas 15.9% (68/429) had 5 or more episodes during follow-up period. When continuous treatment patients were compared with discontinuous treatment patients, living with family members or friends (88.2% (97/110), 78.5% (337/429)), age of first drug use under 25 (61.8% (68/110), 71.3% (306/429)), low urine morphine positive test results (67.3% (74/110), 38.2% (164/429)) and living within 5 kilometers of the MMT clinic (72.7% (80/110), 61.3% (263/429)) were positively associated with higher possibility of continuous treat retention (P < 0.05). Demographics and drug abuse characteristics at enrollment, including gender, age, employment status, family relationship, injection, needle sharing, criminal behavior, contacts with drug users, MMT daily dosage and family members receiving MMT were not significantly associated with treatment retention (P > 0.05).

CONCLUSIONIllicit drug use during the treatment and longer distance travelling to MMT clinic might have negative impact on patients' continuous treatment retention. Mobile MMT vehicles and expanded MMT service sites could be introduced to improve compliance of treatment retention of MMT patients.

Adult ; Female ; Humans ; Longitudinal Studies ; Male ; Methadone ; administration & dosage ; therapeutic use ; Patient Compliance ; statistics & numerical data ; Substance-Related Disorders ; drug therapy ; Treatment Outcome