Objective To study the genomic molecular organization and genogroup of human metapneumovirus(hMPV) infected infants in Guangzhou of China. Methods Primers were designed on the basis of the genomic sequence of hMPV 00-1 strain(AF371337) in the GenBank, and amplify hMPV genomeby RT-PCR. The PCR-products were cloned to T vector and sequenced, the genomic nucleotide sequences were analyzed with the programs Clustal W/X, DNASTAR and MEGA4. 1. Results The cloned strainhMPVgz01 genome is 13 327 bp in length, the genome contains eight open reading frames in the order 3-N-P-M-F-M2-SH-G-L-5. The genomic sequences of hMPVgz01 strain are compared with those of hMPV in GenBank, revealed that the homology with hMPV group A ranges between 92%-97%, homology with group B is 81%, and with avian metapneumovirus group C is 71%, the highest homology is with BJ1887 strain of genogroup A2b. The N, F, G genes of hMPVgz01 strain are compared with those corresponding genes of hMPV subgroups A1, A2, B1, B2, revealed that the highest homology is also with genogroup A2b. Conclusion The complete nucleotide sequence of hMPVgz01 strain isolated from Guangzhou in China is 13 327 bp in length, GenBank accession No. is GQ153651. Comparison of the genomic sequence and three genes of hMPVgz01 strain with those corresponding sequences of hMPV show the highest homology is with genogroup A2b. Sequence and phylogenetic analysis of the hMPVgz01 strain revegled that this isolate belongs to genogroupA2b.

Using the common algorithm and the Mellin algorithm of a continuous wavelet transform, we analyzed the pulse signals of 15 heroin addicts and 15 normal persons. With the use of two algorithms, every pulse signal was processed under 4 scales. From the analyzed results, we found that there was significant difference of wavelet transform coefficients in the time interval 0.2 to approximately 0.4 seconds between the heroin addicts and normal persons. In this paper, the critical parameter used to classify heroin addicts and normal persons is given to every algorithm. The research result of this paper shows that the continuous wavelet transform is really an effective method for processing pulse signals.
Algorithms ; Diagnosis, Computer-Assisted ; Heroin Dependence ; physiopathology ; Humans ; Medicine, Chinese Traditional ; Pulse ; Signal Processing, Computer-Assisted

Objectives: To investigate whether the protective actions of ginsenoside Rb1 (Rb1) on astrocytes are mediated through the Gs-type G-protein-coupled receptor (GPCR-Gs). Methods: Primary astrocyte cultures derived from neonatal mouse brain were used. Astrocyte injury was induced via oxygen-glucose deprivation/re-oxygenation (OGD/R). Cell morphology, viability, lactate dehydrogenase (LDH) leakage, apoptosis, glutamate uptake, and brain-derived neurotrophic factor (BDNF) secretion were assessed to gauge cell survival and functionality. Western blot was used to investigate the cyclic adenosine monophosphate (cAMP) and protein kinase B (Akt) signaling pathways. GPCR-Gs-specific inhibitors and molecular docking were used to identify target receptors. Results: Rb1 at concentrations ranging from 0.8 to 5 μM did not significantly affect the viability, glutamate uptake, or BDNF secretion in normal astrocytes. OGD/R reduced astrocyte viability, increasing their LDH leakage and apoptosis rate. It also decreased glutamate uptake and BDNF secretion by these cells. Rb1 had protective effects of astrocytes challenged by OGD/R, by improving viability, reducing apoptosis, and enhancing glutamate uptake and BDNF secretion. Additionally, Rb1 activated the cAMP and Akt pathways in these cells. When the GPCR-Gs inhibitor NF449 was introduced, the protective effects of Rb1 completely disappeared, and its activation of cAMP and Akt signaling pathways was significantly inhibited. Conclusion Rb1 protects against astrocytes from OGD/R-induced injury through GPCR-Gs mediation.

ObjectiveTo conduct a molecular epidemiological study on human metapneumovirus (hMPV) among pediatric patients in Guangzhou. MethodsA total of 1 840 clinical specimens were obtained from pediatric patients with respiratory infections in Guangzhou Women and Children' s Medical Center in 2010.hMPV was detected by real-time TaqMan RT-PCR in clinical specimens.F gene was amplified and the PCR-products were directly sequenced. ResultsIn 1 840 clinical specimens, 66 werehMPV-positive with a positive rate of 3.59％. hMPV was detected in all specimens except those collected in September and October, and the highest positive hMPV rate occurred in April (6.09％). The F genes of 3 randomly selected strains and hMPVgz01 ( isolated in 2008) were compared with subgroups A1, A2, B1,B2 and C, and the highest homology was with BJ1887 strain of genotype A2b (97％). The F genes of the randomly selected strains and hMPVgz01 were 99％ identical to each other. Sequences and phylogenetics analysis revealed that the epidemical strain in Guangzhou belonged to genotype A2b. ConclusionhMPV is prevalent in spring and summer among children in Guangzhou, and A2b is the predominant genotype.

Objective To study the epidemiology of hand,foot,and mouth disease (HFMD) and the spectrum of serotypes in the other enterovirus (EV) (non-EV-A71 and non-Coxsaekievirus group A 16,CV-A 16) from 2016 to 2017 in Guangzhou,to provide the basis for its treatment,prevention and control.Methods Enteroviruses universal type,EV-A71 and CV-A16 were detected by real time reverse transeription-polymerase chain reaction in the specimens from HFMD suspected patients from 2016 to 2017.The positive specimens of non-EV-A71 and non-CV-A16 were amplified and sequenced based on 5'-untranslated region (UTR) region.The spectrum of serotypes was analyzed with BLAST in NCBI on the basis of 5'-UTR region.Results A total of 25779 specimens from HFMD patients were collected during 2016-2017,16 300 (63.23 ％) of which were positive.The positive rates of EV-A71,CV-A16,non-EV-A71 and non-CV-A16 were 4.57％ (1 178/25 779),12.70％ (3 274/25 779) and 45.96％ (11 848/25779),respectively.The average positive rate of non-EV-A71 and non-CV-A16 in 2017 was 55.68％,which was higher than that in 2016.Sequence analysis showed that there were 16 genotypes in 95 non-EV-A71 and non-CV-A16 positive specimen,including CV-A6,CV-A10,CV-A4,CV-A2,CV-A8,CV-A12,CV-A9,Coxsakievirus B5 (CV-B5),CV-B2,CV-B4,CV-B3,Echovirus 1 (E1),E16,E30,E2 and E18.CV-A6 (26.32％),and CV-A10 (15.79％) were the most common genotypes,followed by CV-A4 (6.32％)、CV-A8(4.21％),and CV-A2 (4.21％).Conclusions The infection rate of EV-A71 is very low during 2016-2017.From April to July 2016,there is a small peak of CV-A16 infection.The non-EV-A71 and non-CV-A16 enterovirus becomes the main causative agent of HFMD during 2016 to 2017.CV-A6 and CV-A10 are the most prevalent pathogens of non-EV-A71 and non-CV-A16 enterovirus.Research and monitoring of CV-A6,CV-A10 as the main non-EV-A71and non-CV-A16 virus should be strengthened.

Objective:To investigate the epidemiology of pathogens of acute respiratory tract infection (ARTI) in children in Guangzhou area.Methods:A total of 13 610 hospitalized children with ARTI in Guangzhou Women and Children′s Medical Center from January 2018 to December 2021 were enrolled. Throat swab specimens were collected, and fluorescent quantitative polymerase chain reaction (PCR) was performed to detect 11 respiratory pathogens, including respiratory syncytial virus (RSV), adenovirus (ADV), parainfluenza virus (PIV), human rhinovirus (HRV), human bocavirus (HBoV), human metapneumovirus (HMPV), enterovirus (EV), influenza A virus (IFA), influenza B virus (IFB), Mycoplasma pneumoniae (MP) and Chlamydia pneumoniae (CP). Grouping according to age (< one year group, one to < three years group, three to < six years group, six to 14 years group) and season. Chi-square test was used for statistical analysis. Results:At least one pathogen was detected in 6 331 cases among 13 610 patients, and the overall positive rate was 46.52%. The detection rates from high to low were as follows: RSV (13.75%(1 872/13 610)), ADV (4.82%(656/13 610)), PIV (4.82%(656/13 610)), MP (4.54%(618/13 610)), HRV (3.39%(462/13 610)), HBoV (2.64%(359/13 610)), HMPV (2.59%(352/13 610)), EV (1.76%(239/13 610)), IFA (1.29%(176/13 610)), IFB (0.90%(122/13 610)) and CP (0.30%(41/13 610)). The positive rate of viral detection showed significant differences among different age groups ( χ2=49.91, P<0.001), and the highest positive rate was in the age group of one to

Despite advances in immunotherapy for the treatment of cancers,not all patients can benefit from programmed cell death ligand 1(PD-L1)immune checkpoint blockade therapy.Anti-PD-L1 therapeutic effects reportedly correlate with the PD-L1 expression level;hence,accurate detection of PD-L1 expression can guide immunotherapy to achieve better therapeutic effects.Therefore,based on the high affinity antibody Nb109,a new site-specifically radiolabeled tracer,68Ga-NODA-cysteine,aspartic acid,and valine(CDV)-Nb109,was designed and synthesized to accurately monitor PD-L1 expression.The tracer 68Ga-NODA-CDV-Nb109 was obtained using a site-specific conjugation strategy with a radiochemical yield of about 95％and radiochemical purity of 97％.It showed high affinity for PD-L1 with a dissociation constant of 12.34±1.65 nM.Both the cell uptake assay and positron emission tomography(PET)imaging revealed higher tracer uptake in PD-L1-positive A375-hPD-L1 and U87 tumor cells than in PD-L1-negative A375 tumor cells.Meanwhile,dynamic PET imaging of a NC1-H1299 xenograft indicated that doxorubicin could upregulate PD-L1 expression,allowing timely interventional immunotherapy.In conclusion,this tracer could sensitively and dynamically monitor changes in PD-L1 expression levels in different cancers and help screen patients who can benefit from anti-PD-L1 immunotherapy.

Objective:To establish the Chinese version of (strength, assistance with walking, rise from a chair, climb stairs and falls, SARC-F) scale using the standardized methods and to validate the reliability and validity for sarcopenia screening among elderly population.Methods:Following the recommended procedure by World Health Organization and European Union Geriatric Medicine Society Sarcopenia Special Interest Group, the translation process included forward translation, expert panel, back-translation, pre-testing and cognitive interviewing to generate the final version. In the pilot study, the test-retest reliability, inter-rater reliability, and internal consistency of the Chinese version of SARC-F scale were assessed. In the diagnostic test for clinical validation, the participants were consecutively recruited from communities and hospitals in Beijing and Tianjin between December 2021 and October 2022. The scale administration, anthropometry, and body composition measurement were conducted by trained investigators. Participants with the SARC-F score ≥ 4 were considered at risk of sarcopenia. Diagnostic tests and receiver operating characteristic curve analysis were performed against the definitions of sarcopenia according to European Working Group on Sarcopenia in Older People (EWGSOP2) and Asian Working Group for Sarcopenia (AWGS2019), and the sensitivity, specificity, positive predictive value, negative predictive value and the area under curve were displayed.Results:The Chinese version of SARC-F scale was approved by the author that the translation has expressed the original meaning correctly. The Chinese version of SARC-F had good test-retest reliability (ICC = 0.914), inter-rater reliability ( r = 0.726), and internal consistency ( α = 0.729). There were altogether 1 882 participants included in the clinical validation. According to the diagnostic criteria of EWGSOP2 and AWGS2019, the Chinese version of SARC-F scale had low sensitivity (13.6% and 16.0%) and positive predictive value (44.6% and 35.4%), high specificity (95.1% and 94.7%) and negative predictive value (79.0% and 86.2%), and moderate AUC of 0.619 and 0.616 (all P < 0.001) for sarcopenia screening. Conclusions:The Chinese version of SARC-F scale was of good reliability and validity. The application of SARC-F in the primary healthcare settings would contribute to the early diagnosis of sarcopenia.

ObjectiveTo observe the neuroprotective effects of ginsenoside Rb₁ on lipopolysaccharide （LPS）-induced neuroinflammation in mice and to preliminarily investigate its mechanism of action. MethodSeventy ICR mice were randomly divided into blank group， model group， dexamethasone sodium phosphate injection group， and low-dose， high-dose ginsenoside Rb₁ groups， with 14 mice in each group. A mouse brain neuroinflammation model was prepared using the LPS dose escalation method， starting with a dose of 1 mg·kg^-1 and administered via intraperitoneal injection every 48 h （every other morning）. Each subsequent dose increased by 2 mg·kg^-1， for a total of 7 injections， culminating in a final dose of 13 mg·kg^-1. The dexamethasone sodium phosphate injection group received an intraperitoneal injection at 5 mg·kg^-1·d^-1. The low-dose and high-dose ginsenoside Rb₁ groups were given intraperitoneal injections at 10 mg·kg^-1·d^-1 and 20 mg·kg^-1·d^-1， respectively， while the blank and model groups received the same volume of normal saline for 14 days. The behavioral activity of LPS mice was observed， anxiety-like behavior was assessed using the Y-maze and elevated plus maze， and brain levels of monocyte chemoattractant protein-1 （MCP-1）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β） were measured by enzyme-linked immunosorbent assay （ELISA）. Neuronal damage of microglia， and the activation status of microglia and astrocytes in the brain were assessed using immunofluorescence staining. The protein expression of phosphatidylinositol-3-kinase （PI3K）， protein kinase B （Akt）， and nuclear factor-κB （NF-κB） in mouse brain were detected by Western blot. ResultCompared with the blank group， the model group showed significantly increased anxiety-like behavior in the Y-maze and elevated plus maze （P<0.05， P<0.01）， significantly elevated levels of MCP-1， TNF-α， and IL-1β in the brain （P<0.01）， a significant decrease in the number of neuronal positive cells in the somatosensory cortex and hippocampus CA1 region （P<0.01）， significant activation of microglia and astrocytes （P<0.01）， and a significant increase in the expression of phosphorylated PI3K， Akt， and NF-κB proteins （P<0.01）. Compared with the model group， the ginsenoside Rb₁ low-dose and high-dose groups showed significantly reduced anxiety-like behavior in the Y-maze and elevated plus maze （P<0.05， P<0.01）， significantly decreased levels of MCP-1， TNF-α， and IL-1β in the brain （P<0.01）， a significant increase in the number of neuronal positive cells in the somatosensory cortex and hippocampus CA1 region （P<0.01）， significant inhibition of microglia and astrocyte activation （P<0.05， P<0.01）， and a significant decrease in the expression of phosphorylated PI3K， Akt， and NF-κB proteins （P<0.05， P<0.01）. ConclusionGinsenoside Rb₁ has neuroprotective effects on LPS-induced inflammation in mice， which may involve the regulation of the PI3K/Akt signaling pathway.

OBJECTIVE To study the pharmacokinetic changes in the plasma and cerebrospinal fluid of bronchial asthma model rats after the complication of Ephedra sinica and Prunus armeniaca. METHODS SD male rats were randomly divided into blank group， model group， E. sinica group （12 g/kg， calculated by raw drug， similarly hereinafter）， P. armeniaca group （6 g/kg） and E. sinica-P. armeniaca drug-pair group （12 g/kg of E. sinica+6 g/kg of P. armeniaca）， with 6 rats in each group. Except for the blank group， the bronchial asthma model was induced by spraying rats in each group with an equal volume mixture of 2% acetylcholine chloride and 0.4% histamine phosphate， once a day， for 7 d. One hour before modeling every time， rats in each group were gavaged with the corresponding drug/normal saline， once a day， for 7 d. After the final administration and provocation of asthma， blood and cerebrospinal fluid collection were performed at different time points. The plasma and cerebrospinal fluid samples were pre-treated （with geranylgeranyl as the internal standard）， and the mass concentrations of ephedrine/pseudoephedrine， methyl ephedrine and amygdalin in both samples were determined by liquid chromatography-tandem mass spectrometry. DAS 2.0 pharmacokinetic software was used to determine the main pharmacokinetic parameters through the non-atrial chamber model and to compare the changes of the pharmacokinetic parameters before and after the combination of the two drugs. RESULTS Compared with E. sinica group， cmax and AUC0-21.33 h （or AUC0-10.67 h） of ephedrine/pseudoephedrine and methyl ephedrine in the plasma and cerebrospinal fluid of rats were significantly reduced in E. sinica-P. armeniaca drug-pair group， while CLZ/F and VZ/F were significantly increased （P＜0.05 or P＜0.01）； tmax of methyl ephedrine in the cerebrospinal fluid was significantly shortened （P＜ 0.05）.Compared with P. armeniaca group， the t1/2 of amygdalin in the plasma of rats in E. sinica-P. armeniaca drug-pair group was significantly shortened， and CLZ/F was significantly increased （P＜0.01）； the tmax of bitter amygdalin in the cerebrospinal fluid was significantly shortened， and the AUC0-10.67 h， CLZ/F， and VZ/F were significantly increased （P＜0.01）. CONCLUSIONS The combination of E. sinica and P. armeniaca accelerates the absorption and elimination of ephedra alkaloids， thus reducing the accumulation of ephedra alkaloids in the bronchial asthma model rats.