Changes in Spectrum of Respiratory Pathogen Infections and Disease Severity Among Children in Hohhot, China: Impact of COVID-19 Prevention Measures.

BACKGROUND This retrospective study evaluated the effects of specific COVID-19 preventive measures, including the use of medical masks, nucleic acid testing, and patient isolation, on respiratory infections, disease severity, and seasonal patterns among children in Hohhot, located in northern China. Understanding these alterations is pivotal in developing effective strategies to handle pediatric respiratory infections within the context of continuous public health initiatives. MATERIAL AND METHODS At the First Hospital of Hohhot, throat swabs were collected from 605 children with community-acquired respiratory between January 2022 and March 2023 for pathogen infection spectrum detection using microarray testing. RESULTS Among the patients, 56.03% were male, and their average age was 3.45 years. SARS-CoV-2 infections were highest between October 2022 and January 2023. Influenza A peaked in March 2023, and other pathogens such as respiratory syncytial virus and influenza B virus disappeared after December 2022. The proportion of mixed infections was 41.94% among SARS-CoV-2 patients, while other pathogens had mixed infection rates exceeding 57.14%. Before December 2022, the mean WBC count for Streptococcus pneumoniae and Haemophilus influenzae was 8.83×109/L, CRP was 18.36 mg/L, and PCT was 1.11 ng/ml. After December 2022, these values decreased significantly. Coughing, difficulty breathing, running nose, and lower respiratory tract infection diagnoses decreased in December 2022, except for SARS-CoV-2 infections. CONCLUSIONS SARS-CoV-2 peaked around November 2022, influenza A peaked in March 2023, and other pathogens like respiratory syncytial virus and influenza B virus were greatly reduced after December 2022. Inflammatory markers and respiratory symptoms decreased after December 2022, except for SARS-CoV-2.

A novel maxillary transverse deficiency diagnostic method based on ideal teeth position.

BACKGROUND: This study proposed a novel maxillary transverse deficiency diagnostic method and evaluated the skeletal Class I and the mild skeletal Class III groups. METHODS: Pre-treatment data from 30 mild skeletal Class III and 30 skeletal Class I patients were collected and uploaded to the Emeiqi Case Management System to design the ideal teeth positions. On these positions, the first bi-molars width was measured at the central fossa and center resistance, the maxillary first bi-premolars width was measured at the central fossa, and the mandibular first bi-premolars width was measured at the distal contact point by Mimics, then width differences of two groups were calculated respectively. RESULTS: At ideal teeth positions, there was no statistically significant difference in the maxillomandibular width in the premolar area between the two groups, but there was in the molar area, and this difference was caused by the difference in mandible width between the two groups. CONCLUSIONS: We proposed a new transverse diagnostic method and found that even the Class I group was not quite up to standard in the molar area on ideal teeth positions, and the Class III group had more severe maxillary transverse deficiency than the Class I group. Meanwhile, the maxillary transverse deficiency in the Class III group was mainly caused by the larger width of the mandible.

Construction of MicroRNA-Target Interaction Networks Based on MicroRNA Expression Profiles of HRV16-infected H1-HeLa Cells.

In the present study we investigated the changes in miRNA levels inhuman rhinovirus 16 (HRV16)-infected cells. A small RNA deep sequencing experiment was performed through next-generation sequencing. In total, 53 differentially expressed miRNAs were confirmed by RT-qPCR, including 37 known miRNAs and 16 novel miRNAs. Interaction networks between differentially expressed miRNAs and their targets were established by mirDIP and Navigator. The prediction results showed that QKI, NFAT5, BNC2, CELF2, LCOR, MBNL2, MTMR3, NFIB, PPARGC1A, RSBN1, TRPS1, WDR26, and ZNF148, which are associated with cellular differentiation and transcriptional regulation, were recognized by 12, 11, or 9 miRNAs. Many correlations were observed between transcriptional or post-transcriptional regulation of an miRNA and the expression levels of its target genes in HRV16-infected H1-HeLa cells.

Genotype and mutation patterns of macrolide resistance genes of Mycoplasma pneumoniae from children with pneumonia in Qingdao, China, in 2019.

OBJECTIVES: This study assessed the incidence and resistance of Mycoplasma pneumoniae (MP) in children in Qingdao, China, in 2019. METHODS: We detected MP infection in 78 pharyngeal swabs from children with pneumonia by qPCR. The RepMP4 element in the P1 adhesin gene, domain V of the 23S rRNA gene, and the L4/L22 ribosomal proteins were amplified by nested PCR. Evolutionary analysis was conducted based on the P1 gene sequence. Resistance mutations in domain V of the 23S rRNA gene and L4/L22 ribosomal proteins were analysed. RESULTS: The incidence of MP infection in children with pneumonia was 59.0% (46/78). The mean duration of MP infection was longer than that of non-MP infection. According to P1 gene sequencing of 21 samples, 12 (57.1%) were type 1 and 9 (42.9%) were type 2. Drug resistance mutations A2063G in domain V of 23S rRNA gene and T508C in L22 were identified from all sequenced MP. However, mutations at positions 2064 and 2617 were not found in this study. C162A mutation appeared in most type 2 samples. A430G mutation appeared in one type 1 sample and in several type 2 samples. T279C mutation in L22 was mostly found in type 2 samples. CONCLUSION: The incidence of MP infection was 59.0% in children with pneumonia in Qingdao in 2019. Type 1 MP infection was slightly more common than type 2, indicating that the genotype of MP is gradually shifting from type 1 to type 2. Macrolide resistance mutation A2063G could be detected in all sequenced MP.

Detection and Analysis of Ebola Virus in Sierra Leone-China Friendship Biosafety Laboratory from March 11 to April 20, 2015.

Ebola virus disease reemerged in Western Africa in 2014. Chinese Center for Disease Control and Prevention dispatched the first Ebola virus (EBOV) detection team to run newly established Sierra Leone-China Friendship Biological Safety Laboratory. The aims of study were to understand epidemiology, clinical manifestations and survival time of EBOV in patient's blood. A total of 913 specimens were tested between March 11 and April 20, 2015. EBOV positivity occurred in 7.37% of the blood and 0.53% in throat swabs. Most commonly reported symptoms of laboratory confirmed patients were intense fatigue, anorexia, and fever. EBOV RNAs persisted in blood for almost 4 weeks and the real-time RT-PCR Ct values showed close correlation with the sampling time after onset.

Good laboratory practices guarantee biosafety in the Sierra Leone-China friendship biosafety laboratory.

BACKGROUND: The outbreak of Ebola virus disease (EVD) in West Africa between 2014 and 2015 was the largest EDV epidemic since the identification of Ebola virus (EBOV) in 1976, and the countries most strongly affected were Sierra Leone, Guinea, and Liberia. FINDINGS: The Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab), a fixed Biosafety Level 3 laboratory in the capital city of Sierra Leone, was established by the Chinese government and has been active in EBOV detection since 11 March 2015. Complete management and program documents were created for the SLE-CHN Biosafety Lab, and it was divided into four zones (the green, yellow, brown, and red zones) based on the risk assessment. Different types of safe and appropriate personnel protection equipment (PPE) are used in different zones of the laboratory, and it fully meets the Biosafety Level 3 laboratory standards of the World Health Organization. CONCLUSION: Good preparedness, comprehensive risk assessment and operation documents, appropriate PPE, effective monitoring and intensive training, together with well-designed and reasonable laboratory sectioning are essential for guaranteeing biosafety.

[Coreceptor usage of human immunodeficiency virus-1 in primary infection through sexual contact transmission].

OBJECTIVE: To evaluate the coreceptor usage characters and evolution of human immunodeficiency virus-1 (HIV-1) primary infection. METHODS: A total of 12 HIV-1 primary infection cases were recruited from a high-risk population for this prospective cohort trial. The blood samples were collected at the earliest time after infection and after a viral setpoint respectively. RNA was extracted from plasma. The env genes were amplified through reverse transcription-PCR (RT-PCR) and nested PCR. And the env fragments were ligated to T vector and then sequenced. The pattern of coreceptor usage was predicted with five different online tools. RESULTS: Among them, 11 were predicted as CCR5 virus transmission and 1 as CXCR4 virus transmission. The viruses in 1 case were detected as a switch of coreceptor usage from CCR5 to CXCR4. The consistency of five different prediction rules was 88.3%. CONCLUSION: Although the CCR5 usage virus is predominant in primary infection through sexual transmission, CXCR4 usage virus can also be detected. A switch of coreceptor usage may happen within the first year post-infection. Adopting multiple rules may improve the efficiency and validity for the prediction of coreceptor usage based on genotypes.

Phosphorylation of Raptor by p38beta participates in arsenite-induced mammalian target of rapamycin complex 1 (mTORC1) activation.

Cell growth is influenced by environmental stress. Mammalian target of rapamycin (mTOR), the central regulator of cell growth, can be positively or negatively regulated by various stresses through different mechanisms. The p38 MAP kinase pathway is essential in cellular stress responses. Activation of MK2, a downstream kinase of p38α, enhances mTOR complex 1 (mTORC1) activity by preventing TSC2 from inhibiting mTOR activation. The p38ß-PRAK cascade targets Rheb to inhibit mTORC1 activity upon glucose depletion. Here we show the activation of p38ß participates in activation of mTOR complex 1 (mTORC1) induced by arsenite but not insulin, nutrients, anisomycin, or H(2)O(2). Arsenite treatment of cells activates p38ß and induces interaction between p38ß and Raptor, a regulatory component of mTORC1, resulting in phosphorylation of Raptor on Ser(863) and Ser(771). The phosphorylation of Raptor on these sites enhances mTORC1 activity, and contributes largely to arsenite-induced mTORC1 activation. Our results shown here and in previous work demonstrate that the p38 pathway can regulate different components of the mTORC1 pathway, and that p38ß can target different substrates to either positively or negatively regulate mTORC1 activation when a cell encounters different environmental stresses.

Inactivation of Rheb by PRAK-mediated phosphorylation is essential for energy-depletion-induced suppression of mTORC1.

Cell growth can be suppressed by stressful environments, but the role of stress pathways in this process is largely unknown. Here we show that a cascade of p38ß mitogen-activated protein kinase (MAPK) and p38-regulated/activated kinase (PRAK) plays a role in energy-starvation-induced suppression of mammalian target of rapamycin (mTOR), and that energy starvation activates the p38ß-PRAK cascade. Depletion of p38ß or PRAK diminishes the suppression of mTOR complex 1 (mTORC1) and reduction of cell size induced by energy starvation. We show that p38ß-PRAK operates independently of the known mTORC1 inactivation pathways--phosphorylation of tuberous sclerosis protein 2 (TSC2) and Raptor by AMP-activated protein kinase (AMPK)--and surprisingly, that PRAK directly regulates Ras homologue enriched in brain (Rheb), a key component of the mTORC1 pathway, by phosphorylation. Phosphorylation of Rheb at Ser 130 by PRAK impairs the nucleotide-binding ability of Rheb and inhibits Rheb-mediated mTORC1 activation. The direct regulation of Rheb by PRAK integrates a stress pathway with the mTORC1 pathway in response to energy depletion.

[Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito].

An artificial intron consisting of the 5'-donor site (from the first intron of the human beta-globin gene) and the 3'-acceptor site (from the intron of an immunoglobulin gene heavy chain variable region) was obtained with a splice overlap extension PCR and was then inserted in frame into the coding sequence of nostructural protein NS1 gene fused to GFP gene in a recombinant mosquito densovirus plasmid p7NS1-GFP. The constructed plasmid was named as p7NS1-Intron-GFP. The plasmid p7NS1-Intron-GFP was co transfected with the helper plasmid pUCA into C6/36 cells, then the packaged recombinant and wild type viruses were purified and recovered. The second-instars of Aedes albopictus larvae were exposed to recombinant and wild type virus mixed stock. The high level GFP expression in C6/36 cells and larvae was observed under fluorescence microscope, indicating that the inserted artificial intron exerted its normal function in self-splicing both in vitro and in vivo. This study laid a foundation for application of an artificial intron in insect cells and development of new strategy for genetic engineering technology of mosqtuito and its pathogens.

SEM and TEM study of the armed male terminal genitalia of the tapeworm Paraechinophallus japonicus (Cestoda: Bothriocephalidea).

For the first time, the ultrastructure of the armed cirrus of an echinophallid cestode, Paraechinophallus japonicus (Yamaguti, 1934), has been studied with the use of scanning and transmission electron microscopy. Two sets of eversible copulatory organs (approximately 300 microm in length and approximately 130 microm in width) are present on the dorsal side of each segment near the lateral margin of the strobila. Except for the terminal portion, the cirrus is covered with large spines (up to 40 mircom long, measured from SEM photomicrographs) composed of 2 parts. The basal portion contains a lobed electron-dense outer region that gives way to a reticular meshwork of electron-dense material. The apical region of the spines, composed of a homogeneous, moderately electron-dense matrix, is slightly curved distally. Spines are covered with a cortical zone. Between the spines, the distal cytoplasm is covered with microvilli of about 1.2 microm in length. The wall of the cirrus sac, which is approximately 500 microm long and approximately 250 microm wide, is composed of 2 layers of muscles, i.e., an internal layer of circular muscles and external longitudinal muscles. Microvilli on the cirrus of P. japonicus are reported for the first time in the Cestoda, whereas the spines on the cirrus may represent a synapomorphy of bothriocephalidean cestodes of the Echinophallidae.

[Improved preparation of pure alive eggs of Schistosoma japonicum].

To prepare a large amount of pure alive Schistosoma japonicum eggs, rabbit was infected with 2000 cercariae and its liver was taken aseptically 38-45 days after infection and homogenized. The homogenate was screened through different sieves(60, 120, 200, 300, 360 meshes per inch respectively), and washed with 1.2% NaCl. The eggs and leftover were then digested with 0.25% trypsin for 2 hours, sieved over 360 meshes per inch and washed with RPMI 1640 medium. The collected eggs reached to (95.1 +/- 6.4)% of live eggs, with a high efficiency.

Alveolar Echinococcus species from Vulpes corsac in Hulunbeier, Inner Mongolia, China, and differential development of the metacestodes in experimental rodents.

Adults of alveolar Echinococcus species with different uterine structures were collected from Vulpes corsac in the Hulunbeier Pasture of Northeastern China in 2001. They were Echinococcus multilocularis Leuckart, 1863 (type No. 3, similar to E. m. multilocularis), with vaselike uterus; Echinococcus cf. sibiricensis Rausch et Schiller, 1954 (type No. 1), with pyriform uterus; and Echinococcus sp. (type No. 2) with spherical uterus at segment top. The metacestode development in rodents also differed among those 3 parasites. In the case of E. multilocularis (type No. 3), many germinal cells grew on the inner surface of early cysts, most of which metastasized into host tissue to form brood vesicles or from the germinal cell layer on the inner surface of the vesicle wall. Cells also had an appearance of proliferating by means of alveolar buds from alveolar tissue that developed outward to form new alveolar foci. In Echinococcus cf. sibiricensis (type No. 1), the formation of alveolar vesicles was due to the metastasizing of germinal tissue into host tissue; protoscoleces grew in the center of alveolar vesicles. In type No. 2 (Echinococcus sp.), the formation of the alveolar vesicle was by multiplication of germinal cell layers on the inner surface of alveolar cysts; protoscoleces grew from the germinal cell layer and mesh in the vesicles. On the basis of uterine structure and on differences in development of metacestodes in experimental rodents, we propose that the 3 types of Echinococcus represent 3 independent species: E. multilocularis, Echinococcus sibiricensis, and Echinococcus sp. (type No. 2-as yet under study).

[Construction of the female subtractive cDNA library and screening of the specific expressing genes].

OBJECTIVE: To screen the Schistosoma japonicum female specific expressing genes. METHODS: S. japonicum adult worms were collected from the rabbits' vein after six-week infection by affusing method. The adult worms were stabilized by RNA-later liquid, the male and female worms were carefully separated with nipper. The high quality total RNA was extracted and mRNA was obtained after purification. Double stranded cDNAs were synthesized after reverse transcription. Female subtractive (female as tester, male as driver) and male subtractive (male as tester, female as driver) cDNA libraries were constructed. The differentially expressed genes were further screened by dot-blot hybridization. The clones were selected and sequenced, which showed apparently higher signals when hybridizing with the female subtracting male probes, than those signals when hybridizing with the male subtracting female probes. The homology of these sequences was searched with BLAST program. The semi-quantitative PCR was applied to test the differential gene expression in female and male adult worms. RESULT: Female subtracting male and male subtracting female cDNA libraries were constructed with SSH technique. After dot-blot hybridization, 50 clones were tested to be the potential female differentially expressed genes and were sequenced. 42 expressing sequence tags (ESTs) were received. After bioinformatics analysis, 17 fragments (about 40.5%) showed high identity with the S. japonicum egg-shell protein genes, 17 sequences (about 40.5%) were highly homologous to unknown S. japonicum genes and partly homologous to female specific 800 protein. 8 fragments (about 19.0%) showed high identity with other S. japonicum unknown genes. The fragments in clones of 577, 579, 668, 695, 720, and 708 were tested by RT-PCR to be the differentially expressed genes in female adult worms using S. japonicum actin gene as the internal standard. These fragments were highly homologous to S. japonicum egg shell protein gene AY222885, AY222895, AB017097, AF519182, M32281, and S. japonicum unknown gene AY813556 respectively. CONCLUSION: SSH is essential to screen the differentially expressed genes of S. japonicum female worms. A number of female specific genes have been found by this method.

A new pseudophyllidean from a freshwater fish, Gambusia affinis, in China.

Coelobothrium gambusiense n. sp. (Bothriocephalidae) was collected and described from the intestine of the freshwater fish Gambusia affinis (Baird and Girard) (Poeciliidae) in Fujian Province, Peoples' Republic of China. It is the first record of Coelobothrium in China. The parasite closely resembles Coelobothrium monodi Dollfus, 1970, from Capoeta damascina (Valenciennes, Cyprinidae) in Iran and Coelobothrium oitense Kugi and Matsuo, 1990, from Tribolodon hakonensis (GUnther, Cyprinidae) in Japan in general morphological characters, the scolex, and the incomplete proglottids. The third species of Coelobothrium is distinguished from its congeners by its much shorter strobila, presence of a neck, a bilobed ovary instead of a transversely elongated ovary, larger eggs, different final host and locality, and other morphological characters.

Saccocoelium megasacculum n. sp., a new haploporid trematode from Liza carinatus in the Taiwan strait.

Saccocoelium megasacculum n. sp. (Digenea: Haploporidae) was collected from the intestine of the mugilid fish. Liza carinatus (Cuvier and Valenciennes), in the Taiwan Strait. It is the first record of Saccocoelium in China. The parasite most closely resembles Saccocoelium obesum Looss, 1902 and Saccocoelium tensum Looss, 1902 in general morphology and body size, but it is easily distinguished from them in having a larger hermaphroditic sac in relation to body size; larger eggs; smaller pharynx, testis, ovary, and vitellaria; and a uterine seminal receptacle instead of a true seminal receptacle.

Parabothriocephalus psenopsis n. sp. (Eucestoda: Pseudophyllidea) in Psenopsis anomala from the Taiwan Strait, China.

Palabothriocephalus psenopsis n. sp. (Eucestoda: Pseudophyllidea) is described from the gastrointestinal tract of Psenopsis anomala caught off the coast of Xiamen, China. The new species most closely resembles, but differs from, Parabothriocephalus segmentatus in its possession of muscular globular appendages on the posterior margin of the proglottids, a limited proglottid number (9-13), and the shorter strobila (7.6-13.2 mm vs. 165 mm). In addition, the uterus of P. psenopsis is strongly coiled, whereas that of P. segmentatus is S shaped; P. segmentatus has a spherical expansion in the middle of the vagina, whereas that of P. psenopsis does not. Finally, P. psenopsis differs from Parabothriocephalus gracilis. Parabothriocephalus sagitticeps, and Parabothriocephalus macruri by the posterolateral expansion of the proglottids.