Objective To investigate the efficacy of extracorporeal shock wave therapy(ESWT)combined with ultrasound-guided corticosteroid injection(CSI)for the treatment of primary frozen shoulder(PFS).Methods Ninety-nine patients with PFS who visited the pain department of the Affiliated Hospital of Xuzhou Medical University between April 2024 and July 2024 were enrolled and randomly divided into three groups according to the randomized number table method:ESWT group(T group),CSI group(I group),and combined treatment group(TI group),with 33 patients in each group.Visual analogue scale(VAS)scores,shoulder range of motion(SROM),and Constant-Murley shoulder scores(CMS)were recorded before treatment and at 1,4,8,and 12 weeks post-treatment.Additionally,the patients'Ascens Insomnia Scale(AIS)scores were recorded before treatment and 1 month after treatment.The occurrence of adverse effects and the use of remedial medications during the treatment period were also documented.Results Compared with pre-treatment,VAS scores decreased,and SROM and CM scores improved at all time points after treatment in all three groups(P<0.05).AIS scores also decreased in all three groups at 1 month post-treatment(all P<0.05).Intergroup comparisons revealed that the TI group exhibited signifi-cantly lower VAS pain scores,greater SROM(forward flexion and backward extension),and higher CM scores at 4,8,and 12 weeks post-treatment compared to the T and I groups(Bonferroni-corrected P<0.05).No statistically significant differences were observed between the T and I groups for these measures(Bonferroni-corrected P>0.05).Additionally,there were no statistically significant differences in AIS scores or adverse effects occurrence among the three groups at 1 month post-treatment(P>0.05).Conclusion The combined treatment demonstrated greater efficacy compared to trigger point extracorporeal shock wave therapy alone and periapical steroid injection alone,resulting in significant improvement in the patient's clinical symptoms and quality of life.

Objective To investigate the efficacy of extracorporeal shock wave therapy(ESWT)combined with ultrasound-guided corticosteroid injection(CSI)for the treatment of primary frozen shoulder(PFS).Methods Ninety-nine patients with PFS who visited the pain department of the Affiliated Hospital of Xuzhou Medical University between April 2024 and July 2024 were enrolled and randomly divided into three groups according to the randomized number table method:ESWT group(T group),CSI group(I group),and combined treatment group(TI group),with 33 patients in each group.Visual analogue scale(VAS)scores,shoulder range of motion(SROM),and Constant-Murley shoulder scores(CMS)were recorded before treatment and at 1,4,8,and 12 weeks post-treatment.Additionally,the patients'Ascens Insomnia Scale(AIS)scores were recorded before treatment and 1 month after treatment.The occurrence of adverse effects and the use of remedial medications during the treatment period were also documented.Results Compared with pre-treatment,VAS scores decreased,and SROM and CM scores improved at all time points after treatment in all three groups(P<0.05).AIS scores also decreased in all three groups at 1 month post-treatment(all P<0.05).Intergroup comparisons revealed that the TI group exhibited signifi-cantly lower VAS pain scores,greater SROM(forward flexion and backward extension),and higher CM scores at 4,8,and 12 weeks post-treatment compared to the T and I groups(Bonferroni-corrected P<0.05).No statistically significant differences were observed between the T and I groups for these measures(Bonferroni-corrected P>0.05).Additionally,there were no statistically significant differences in AIS scores or adverse effects occurrence among the three groups at 1 month post-treatment(P>0.05).Conclusion The combined treatment demonstrated greater efficacy compared to trigger point extracorporeal shock wave therapy alone and periapical steroid injection alone,resulting in significant improvement in the patient's clinical symptoms and quality of life.

Objective To explore the presence and potential interactions of microbes and bacteriophages in the blood of healthy individuals by employing in-depth bioinformatics mining to analyze the structure and function of the blood microbi-ome ecosystem.Methods Blood plasma samples from 1 600 voluntary blood donors collected at Mianyang Central Blood Station from 2012 to 2018 were subjected to DNA extraction and library construction.High-throughput sequencing was con-ducted using the Illumina HiSeq 4500 platform,followed by extensive bioinformatics analysis.Microbial abundance in blood samples was analyzed using metagenomic analysis software such as Bowtie2,Trimmomatic and Kraken.Subsequent phage-ome analysis included sequence quality control,assembly,identification,clustering and functional annotation using software such as Megahit,geNomad,CheckV and eggNOG-mapper.Phylogenetic trees,species annotation and host analysis and pre-diction for the identified blood bacteriophages were constructed using iTOL,BLAST and PhaBOX software.Results Met-agenomic sequencing identified microbes across 36 phyla,151 orders,338 families,338 genera and 3 757 species in the plasma samples.At the species level,the most abundant species included Bacillus cereus,Lactobacillus murinus,L.johnso-nii,Faecalibacterium prausnitzii,B.thuringiensis,L.reuteri,Cutibacterium acnes,Dietzia sp.JS16-p6b,Mycoplasma hyo-rhinis,M.hyopneumoniae and Staphylococcus aureus.Through phageome analysis,202 viral Operational Taxonomic Units(vOTUs)were identified,revealing 24 types of bacteriophages.Host analysis using the viral host database completed mat-ches for 15 potential bacteriophage hosts,including Stenotrophomonas maltophilia,Rhodoferax lacus,Pseudoalteromonas marina,Thalassotalea loyana,Vibrio alginolyticus,V.tasmaniensis,V.vulnificus,Pseudomonas sp.,Agrobacterium sp.ST15.13/040,Enterococcus gallinarum,Flavobacterium sp.,Thermotoga naphthophila,Chryseobacterium sp.RU33C,L.acidipiscis and Neisseria mucosa.Conclusion The study of the healthy human blood microbiome and phageome reveals the presence of microbes and phages in the blood,which may have profound impacts on human health.

Cancer stands as one of the predominant causes of mortality globally, necessitating ongoing efforts to develop innovative therapeutics. Historically, natural products have been foundational in the quest for anticancer agents. Bulbocodin D (BD) and Bulbocodin C (BC), two bibenzyls derived from Pleione bulbocodioides (Franch.) Rolfe, have demonstrated notable in vitro anticancer activity. In human lung cancer A549 cells, the IC_50s for BD and BC were 11.63 and 11.71 μmol·L^-1, respectively. BD triggered apoptosis, as evidenced by an upsurge in Annexin V-positive cells and elevated protein expression of cleaved-PARP in cancer cells. Furthermore, BD and BC markedly inhibited the migratory and invasive potentials of A549 cells. The altered genes identified through RNA-sequencing analysis were integrated into the CMap dataset, suggesting BD's role as a potential signal transducer and activator of transcription 3 (STAT3) inhibitor. SwissDock and MOE analyses further revealed that both BD and BC exhibited a commendable binding affinity with STAT3. Additionally, a surface plasmon resonance assay confirmed the direct binding affinity between these compounds and STAT3. Notably, treatment with either BD or BC led to a significant reduction in p-STAT3 (Tyr 705) protein levels, regardless of interleukin-6 stimulation in A549 cells. In addition, the extracellular signal-regulated kinase (ERK) was activated after BD or BC treatment. An enhancement in cancer cell mortality was observed upon combined treatment of BD and U0126, the MEK1/2 inhibitor. In conclusion, BD and BC emerge as promising novel STAT3 inhibitors with potential implications in cancer therapy.
Humans ; Lung Neoplasms/metabolism* ; STAT3 Transcription Factor/metabolism* ; Antineoplastic Agents/chemistry* ; A549 Cells ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation

Objective:To learn about the characteristics of natural foci and the spatial distribution of mosquitoes and ticks in Yadong County, Tibet Autonomous Region (Tibet for short).Methods:The eco-geographical characteristics, demographic information, agricultural and animal husbandry data of the natural foci in Yadong County, Tibet were collected from the Yadong County 2020 statistical yearbook, vector data were collected from the Yadong County Center for Disease Control and Prevention, and the data of the meteorological station in Yadong County from 2010 to 2021 were collected from the World Weather Network. In July 2021, a total of 20 mosquito and tick sampling points were selected for field investigation according to the distribution characteristics of vegetation and animal husbandry of Yadong County. The light trap method was used to trap mosquitoes, and the manual inspection and flag dragging method were used to catch ticks. The spatial distribution characteristics of mosquitoes and ticks were analyzed.Results:The natural epidemic foci in Yadong County, Tibet, were high in the north and low in the south, with an elevation difference of about 5 000 m. It was rich in water resources, and the average runoff of Yadong River was 20.1 m 3/s. The climate in the north was cold and dry, and the south was mild and humid. From 2010 to 2021, the annual average station air pressure in Yadong County was 452.8 mmHg (1 mmHg = 0.133 kPa), the maximum monthly average temperature, precipitation and relative humidity of air were 8.7 ℃, 134.5 mm and 81.3%, respectively, all in July. A total of 6 897 diptera insects were attracted by the light trap method, all of which were non-blood-sucking mosquitoes. The total density was 163.77 pieces/(lamp·h), the dominant population was Muscaridae, accounting for 89.69% (6 186/6 897). In different sampling areas, Xiayadong Township collected the most mosquitoes, accounting for 67.17% (4 633/6 897); the others were Yadong County and Shangyadong Township, accounting for 27.36% (1 887/6 897) and 5.47% (377/6 897), respectively. A total 2 014 host animals were examined, and 23 parasitic ticks were caught, of which 20 were of the genus Ixodes and 3 were of the genus Haemophilus. No free ticks were caught in all ticks sampling points. Conclusions:The climate and ecological environment of the natural foci in Yadong County, Tibet have obvious vertical gradient. Mosquitoes and ticks are active, but the density is not high, the density of mosquitoes is generally high in the south and low in the north.

OBJECTIVE: To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. METHODS: We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. RESULTS: This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. CONCLUSION By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans ; COVID-19 ; CRISPR-Cas Systems ; Genotype ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2/genetics* ; RNA ; COVID-19 Testing

Objective:To investigate the molecular epidemiologic and clinical characteristics of norovirus (NoV) in hospitalized children with acute gastroenteritis (AGE) in Tianjin to provide theoretical basis for clinical diagnosis and treatment.Methods:Fecal specimens were collected from children with AGE in Tianjin Children′s Hospital from August 2017 to July 2020. NoV was detected by real-time fluorescence reverse transcription-polymerase chain reaction (RT-PCR). Partial sequence of the capsid protein VP1 of positive samples was amplified by conventional RT-PCR. The products were sent for sequencing and genotyping based on the sequence. As the same time, the clinical symptoms were compared among children infected with different NoV genotypes.Results:Among the 6 432 specimens, 1 703 (26.5%) were positive for NoV. Sequence analysis showed that 761 identified NoV strains could be divided into 7 genotypes. Genotypes GII.4, GII.3, GII.2, GII.17, GII.1, GII.13 and GII.6 accounted for 55.5% (422/761), 36.7% (279/761), 4.9% (37/761), 0.9% (7/761), 0.8% (6/761), 0.8% (6/761) and 0.5% (4/761), respectively. All of the GII.4 belonged to GII.4 Sydney 2012. There was also a significant difference in the seasonal distribution of children infected with GII.4 and other genotypes ( χ2=103.53, P<0.001). Children infected with GII.4 NoV were more likely to have diarrhea, vomiting and dehydration ( χ2=8.42, P=0.004; χ2=20.39, P<0.001; χ2=4.99, P=0.025). Conclusions:GII.4 and GII.3 were mainly genotypes of NoV infection in Tianjin from 2017 to 2020 and genotypes were diverse. In addition, children with GII.4 NoV were more likely to suffer from diarrhea, vomiting and dehydration.

Objective:To understand the pathogen infection and epidemiological characteristics of children with diarrhea in Tianjin.Methods:Stool samples from 1 466 children with diarrhea in Tianjin Children's Hospital from August 2017 to July 2018 were collected, and all samples were tested for five intestinal-related pathogens (norovirus, rotavirus, Clostridium difficile toxin, adenovirus, and astrovirus). Results:Among the 1 466 samples, 627 samples were positive for nucleic acid detection of intestinal pathogens, with a positive rate of 42.8%. The detection rate of norovirus was the highest (26.3%), followed by rotavirus (15.3%), Clostridium difficile toxin (4.6%), adenovirus (4.1%), and astrovirus (1.84%). The infection has obvious seasonality. The positive detection rates of the five pathogens were similar among children of different sexes, and only the positive detection rates of norovirus and rotavirus were statistically significant among different ages ( P<0.05). There were 110 cases of mixed infection, and the mixed infection of norovirus and rotavirus was the most common, with a total of 37 cases. Conclusions:The pathogen spectrum of infant infectious diarrhea in Tianjin is complex and diverse, and the main pathogens are norovirus and rotavirus.

Objective:To analyze the pathogenic bacteria and epidemiological characteristics in children with respiratory tract infection in Tianjin area.Methods:Retrospective case analysis was performed on 2 392 hospitalized children in the wards of respiratory diseases, intensive care unit and special care ward of Tianjin Children′s Hospital from June 2018 to May 2019. Thirteen pathogenic bacteria in deep sputum and bronchoalveolar lavage fluid samples were detected by loop-mediated isothermal amplification. The laboratory data and clinical characteristics of the infected children were analyzed, and the comparison between groups was performed by t test or χ 2 test. Results:Among 2 392 cases, 1 407 were males and 985 females. There was no significant difference in the detection rate between males and females (72.5% (1 020/1 407) vs.74.2% (731/985), χ 2=0.87, P=0.35). A total of 1 751 strains and 12 kinds of positive respiratory pathogens were detected, with a detection rate of 73.2%. Among them, 913 (38.2%) strains were Mycoplasma pneumoniae (MP), 514 (21.5%) were Streptococcus pneumoniae (Sp), 381 (15.9%) were Methicillin-resistant Staphylococcus aureus (MRSA) and 279 (11.7%) were Hemophilus influenzae (Hi). There was significant difference in the detection rate of pathogens among different age groups (χ2=83.67, P<0.01). The positive rate of alveolar lavage fluid group was higher than that of deep sputum fluid group [81.6% (614/752) vs. 69.3% (1 137/1 640), χ 2=39.89, P<0.01]. The length of hospital stay of children infected with different pathogens was significantly different (all P<0.01). There was significant difference in duration of fever among children infected with different pathogens (χ2=228.69,103.56, 3.96, 27.38,24.50,41.66, all P<0.05). There were 63 (7.7%) cases of atelectasis, 260 (31.9%) cases of pleurisy and 120 (14.7%) cases of pleural effusion in MP children. Children with Sma were most likely to involve the heart system (2/9), and children with Eco infection had a higher incidence of complications such as those of blood (3/19), urinary (2/19), digestive systems(4/19), systemic inflammatory response syndrome and sepsis (1/19). Conclusions:The main bacterial pathogens of respiratory tract infection in children in Tianjin were MP, Sp, MRSA and Hi. It is suggested that clinicians should not only pay attention to the respiratory symptoms of children, but also pay attention to the complications caused by bacterial pathogen infection, so as to prevent the deterioration of the disease and improve the prognosis.

Objective:To investigate the molecular epidemiological characteristics of norovirus (NoV) in hospitalized children with sporadic acute gastroenteritis in Tianjin in 2019.Methods:Fecal specimens and clinical data were collected from 3 116 hospitalized children with sporadic acute gastroenteritis possibly caused by viral infection in Tianjin Children′ Hospital between January and December, 2019. Real-time quantitative PCR was used to detect NoV. Partial sequences of RNA-dependent RNA polymerase (RdRp) and capsid genes of NoV were amplified by RT-PCR. Sequence alignment and phylogenetic analysis were performed for further analysis.Results:Among the 3 116 specimens, 809 (26.0%) were positive for NoV. There were significant differences in NoV detection rate between different age groups ( P=0.000), and the highest NoV detection rate (31.6%) was observed in the age group of 7-12 months. Moreover, the detection rate of NoV varied with seasons ( P=0.000), and the NoV detection rate was highest in winter (39.0%). Based on the sequence analysis of RdRp and capsid genes, 286 identified NoV strains belonged to six genotypes, which were GⅡ.P12-GⅡ.3, GⅡ.P16-GⅡ.2, GⅡ.P17-GⅡ.17, GⅡ.Pe-GⅡ.2, GⅡ.Pe-GⅡ.3 and GⅡ.Pe-GⅡ.4. The predominant genotype was GⅡ.Pe-GⅡ.4 Sydney 2012 (61.2%), followed by GⅡ.P12-GⅡ.3 (33.6%, 96/286), GⅡ.Pe-GⅡ.3 (2.4%, 7/286), GⅡ.P16-GⅡ.2 (2.1%, 6/286), GⅡ.Pe-GⅡ.2 (0.3%, 1/286) and GⅡ.P17-GⅡ.17 (0.3%, 1/286). Patients carrying the NoV of GⅡ.Pe-GⅡ.4 Sydney 2012 genotype were likely to suffer from vomiting than those positive for NoV of GⅡ.P12-GⅡ.3 genotype. Conclusions:NoV was an important pathogen causing acute gastroenteritis in children. GⅡ.Pe-GⅡ.4 Sydney 2012 and GⅡ.P12-GⅡ.3 were the major genotypes of NoV in hospitalized children with sporadic acute gastroenteritis in Tianjin in 2019.