ObjectiveTo analyze the infection characteristics of human respiratory syncytial virus (HRSV) among children hospitalized with acute lower respiratory tract infection (ALRTI) in a specialized pediatric hospital in Shanghai, so as to provide evidence-based support for optimizing the prevention and control strategies and clinical diagnosis and treatment of respiratory tract infections in children in this region. MethodsA retrospective analysis was performed to the clinical and etiological data of 29 260 children hospitalized for ALRTI in Shanghai Children’s Hospital from January 2021 to December 2024. HRSV and 12 other common respiratory pathogens were detected with multiplex polymerase chain reaction (PCR) and capillary electrophoresis. Demographic and clinical data were collected for statistical analyses. A total of2 412 cases with positive HRSV were divided into the severe group and the non-severe group. Clinical characteristics between the two groups were compared using the Mann-Whitney U test and the chi- square (χ²) test. Additionally, the related influencing factors of severe HRSV infection were explored. ResultsThe overall positivity rate of HRSV from 2021 to 2024 was 8.24% (2 412/29 260), with statistically significant differences observed across the four years (χ²=389.42, P<0.001). The highest positivity rate was in 2021 (14.76%), with a high prevalence throughout the year. In 2022, when non-pharmaceutical interventions (NPIs) were implemented, the HRSV positivity rate was the lowest (4.93%), with a winter-dominant epidemic pattern. In 2023, after the NPIs were lifted, the HRSV positivity rate showed a slight rebound (8.14%), presenting a double-peak pattern. In 2024, the HRSV positivity rate slightly decreased compared to that in 2023 (6.29%), exhibiting a winter and spring-dominant epidemic pattern. Among the hospitalized children with ALRTI, the HRSV positivity rate in males (8.85%) was higher than that in females (7.51%), and the difference was statistically significant (χ²=17.33, P<0.001). Age distribution showed that 82.26% (1 984/2 412) of HRSV infections occurred in children aged 3 years old and below. Besides, as age increased, the infection rate of HRSV showed a gradually decreasing trend (P<0.001). Among the 2 412 children with HRSV infection, the proportion of severe cases was 22.31% (538/2 412), while the non-severe cases accounted for 77.69% (1 874/2 412). Compared with non-severe cases, severe cases were more frequently presented with high fever, longer duration of wheezing, as well as higher rates of underlying diseases or co-infection with Mycoplasma pneumoniae (P<0.001). ConclusionThe prevalence intensity of HRSV varied yearly from 2021 to 2024. After the removal of NPIs in 2023, a slight rebound with a double-peak epidemic pattern was observed. HRSV remained a common pathogen in children hospitalized for ARLTI, and children aged 3 years old and below constituted the highest proportion for infection. Compared with non-severe cases, those with severe HRSV infections were more prone to presenting with high fever and a longer duration of wheezing. Children with positive HRSV who had underlying diseases or co-infection with Mycoplasma pneumonia were more likely to develop severe conditions.

The irrational use of Chinese patent medicines （CPM） is becoming more and more prominent， which makes the demand for clinical practice guidelines of CPM gradually increase. In order to make domestic scholars understand the latest developments and existing problems of the CPM guidelines， and promote its development， this paper introduced the concept of CPM guidelines， summarized the characteristics of the two development modes， namely “taking CPM as the key” and “taking disease/syndrome as the key”， and analyzed the current methodological status of developing and reporting CPM guidelines. Based on the existed problems， three suggestions have been put forward to optimize the quality of CPM guidelines， which were clarifying the target users and scope of CPM guidelines， establishing an open and transparent mechanism of the personnel involvement and process steps， and formulating implementable and operable recommendations for the use of CPM.

The identification of clinical questions for clinical practice guidelines of Chinese patent medicine （CPM） is important for subsequent evidence retrieval， evaluation of evidence quality， formation of recommendations. This paper described a methodological proposal for the identification of clinical questions for CPM guidelines to highlight the characteristics of Chinese patent medicine and reflect its effect in specific stage of the disease. Considering four aspects， namely， the drug of Chinese patent medicine （D）， the specific disease stage （S）， comparison （C）， and specific outcome （O）， DSCO framework has been proposed to formulate the clinical questions. Multi-source information through scientific research， policy or standard documents， and clinical data are suggested for collecting clinical questions， and clear selection criteria should be set to finalize the clinical questions to be addressed by the guideline. In addition， the above process needs to be transparently and publicly reported in order to ensure the clarity and completeness of the guidelines.

Objective:To observe the expression changes of microRNA(miR)-122 in liver tissue of rats infected with Clonorchis sinensis and its correlation with expression level of inflammatory cytokines. Methods:Totally 24 SPF grade Wistar male rats were selected and randomly divided into a control group (100 μl physiological saline gavage), a 4-week infection group (100 Clonorchis sinensis metacercariae gavage), and an 8-week infection group (100 Clonorchis sinensis metacercariae gavage) based on body weight (100-120 g) using a random number table method, with 8 rats in each group. Starting from the third week of infection, rat feces were collected and directly smeared with physiological saline for identification of Wistar rat animal models infected with Clonorchis sinensis. After 4 and 8 weeks of infection, the rats in the 4- and 8-week infection groups were euthanized, while 4 rats in the control group were euthanized, respectively. The heart blood and left lobe liver tissue and serum samples were collected from each group of rats. Using hematoxylin-eosin (HE) staining to observe liver pathological damage under the light microscope, real-time fluorescence quantitative PCR to detect the expression level of miR-122 in liver tissue, and Luminex 200 liquid suspension chip to detect the expression levels of serum inflammatory cytokines [tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6 (IL-6)]. The correlation between miR-122 and inflammatory cytokines was analyzed using Pearson correlation. Results:Under the light microscope, the morphology of hepatocytes in control group was normal, and no inflammatory cell infiltration was observed. There was inflammatory cells such as lymphocyte, eosinophil and other inflammatory cell infiltration around the portal area in the 4-week infection group. The hepatocytes of the 8-week infected rats were arranged in a disordered manner, with varying degrees of swelling, loose and lightly stained cytoplasm, and some hepatocytes showed watery degeneration; additionally, bile duct dilation and thickening of the bile duct wall were observed in the liver tissue. There were statistically significant differences of liver miR-122 (1.00 ± 0.32, 2.57 ± 0.60, 3.63 ± 1.63), serum TNF-α [(0.14 ± 0.06), (0.43 ± 0.09), (0.61 ± 0.10) ng/ml], and IL-6 expression levels [(0.03 ± 0.01), (1.06 ± 0.24), (1.48 ± 0.33) ng/ml] in control group, 4- and 8-week infection groups ( F = 13.36, 69.99, 82.23, P < 0.001). There was no statistically significant difference in expression level of IL-1β between different groups ( F = 2.15, P = 0.141). The Pearson correlation analysis showed that the expression level of miR-122 was positively correlated with the expression levels of inflammatory cytokines TNF-α and IL-6 ( r = 0.67, 0.80, P < 0.001). Conclusion:Clonorchis sinensis infection can increase the expression of miR-122 in the host liver tissue, and the miR-122 is closely related to the expression levels of inflammatory cytokines TNF-α and IL-6.

Objective:To analyze cytokine pattern of systemic onset juvenile idiopathic arthritis (SOJIA) combined with macrophage activation syndrome (MAS) in children, and study the early diagnostic value in MAS.Methods:The clinical data of 157 children with SOJIA from January 2013 to March 2018 in Taizhou Hospital of Zhejiang Province were retrospectively analyzed. Among them, SOJIA combined with MAS was in 15 cases (SOJIA combined with MAS group), and simple SOJIA was in 142 cases (simple SOJIA group). The peripheral blood levels of interleukin (IL)-2, IL-4, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were measured by flow cytometry cytometric beads array. The characteristics of cytokine pattern was analyzed.Results:The IL-10 and IFN-γ in SOJIA combined with MAS group were significantly higher than those in simple SOJIA group: 40.5 (7.9, 236.9) ng/L vs. 4.1 (2.0, 98.7) ng/L and 55.8 (18.5, 500.0) ng/L vs. 4.4 (1.4, 30.1) ng/L, and there were statistical differences ( P<0.01); there were no statistical difference in IL-2, IL-4, IL-6 and TNF-α between 2 groups (P>0.05). Pearson correlation analysis result showed that IL-10 was positive correlated with IFN-γ in SOJIA children with MAS ( r = 0.638, P = 0.011). Receiver operating characteristic curve analysis result showed that the area under the curve (AUC) of IFN-γ in predicting MAS was 0.991, 95% CI 0.974 to 1.000, the optimal critical value was 18.45 ng/L, the sensitivity was 92.5%, and the specificity was 95.1%; the AUC of IL-10 in predicting MAS was 0.944 (95% CI 0.893 to 0.996), the optimal critical value was 7.75 ng/L, the sensitivity was 91.7%, and the specificity was 81.7%. Conclusions:The significant increased IL-10 and IFN-γ is helpful for the early diagnosis MAS in children with SOJIA.

Objective:To investigate the prevalence and molecular characteristics of Escherichia coli ( E. coli) producing a novel Shiga toxin 2k subtype in goat in Lanling county, Shandong province. Methods:In November 2019, 512 goat fecal samples were collected from different households in Lanling county, Shandong province. After enriched with EC broth, stx-positive samples were detected by PCR and inoculated in CHROMagar? ECC agar and CHROMagar? STEC agar. The whole genomes of stx-positive strains were sequenced. Based on the genomic senquences, the stx subtype, serotype, multi-locus sequence type and virulence genes of each strain were analyzed. Results:Eighty-six strains of Shiga toxin-producing E. coli (STEC) were isolated from 512 goat fecal samples. Five stx subtypes were identified and 37 strains were positive for stx2k. The 86 STEC strains belonged to 20 O∶H serotypes and 18 different sequence types (STs). Conclusions:STEC strains circulating in goats in Lanling county, Shandong province were heterogeneous in stx subtypes, serotypes and virulence gene profiles, and a certain proportion of strains producing a novel Shiga toxin 2k subtype were detected.

Objective To evaluate the efficacy of wax therapy for diabetic peripheral neuropathy, in order to provide suggestions for treatment and prevention.Methods Cochrane Library, JBI, PubMed, EMbase, CBM, CNKI, WANFANG DATA, VIP were searched by computer from inception to October, 2016 for randomized control trails in wax therapy for diabetic peripheral neuropathy. Two researchers searched and selected articles independently and the quality was assessed in accordance with Cochrane Manual. The articles were analyzed with Review Manager 5.3, and the evidence quality was assessed with GRADE profiler 3.6.1 software.Results A total of 8 randomized controlled trials were included. The number of literature quality grade A of original research is 3, while the number of literature quality grade B of original research is 4,and the number of literature quality grade C of original research is 1. Meta-analysis showed that the sensory nervus peronaeus conduction velocities improved by wax therapy for diabetic peripheral neuropathy patients,[MD=2.51, 95%CI(1.35,3.66), P<0.01]. By the GRADE system grading, the evidence quality is moderate.Conclusions The effect of wax therapy for diabetic peripheral neuropathy can improve the sensory nurve conduction velocities and speed of blood circle, so as to prevent ulcer of diabetic feet. Therefore, the clinical curative effects of the therapy require large-sample and high quality studies to confirm.

OBJECTIVETo investigate the serum levels of Th1/Th2 cytokines in children with non-systemic juvenile onset idiopathic arthritis (non-SOJIA).

METHODSClinical data of 41 children with non-SOJIA, including 11 cases of polyarthritis, 10 cases of oligoarthritis and 20 cases of enthesitis related JIA (ERA), admitted in Children's Hospital of Zhejiang University School of Medicine during November 2012 and May 2015 were retrospectively analyzed. Serum levels of Th1/Th2 cytokines including IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ were measured by flow cytometry in patients with non-SOJIA, and compared with those in patients with SOJIA (SOJIA group, n=85) and healthy children (control group, n=202); their correlations with erythrocyte sedimentation rate and C reactive protein and CRP were analyzed.

RESULTSCompared with the healthy control group, serum levels of IL-2, IL-6 and IFN-γ were significantly increased in patients with non-SOJIA (2.9 vs. 2.6 pg/mL, 9.9 vs. 6.4 pg/mL, 6.3 vs. 5.1 pg/mL, allP<0.05),while levels of TNF-α and IL-10 were significantly decreased (2.7 vs. 3.9 pg/mL, 2.9 vs. 7.1 pg/mL, both P<0.01). Compared with the SOJIA group, serum levels of IL-6 and IL-10 were significantly decreased in patients with non-SOJIA (9.9 vs. 33.5 pg/mL, 2.9 vs. 4.1 pg/mL, both P<0.01), while levels of IL-4 and IL-10 were significantly increased (3.1 vs. 2.3 pg/mL, 6.3 vs. 4.4 pg/mL, both P<0.05). Serum levels of IL-6 in patients with polyarthritis or ERA were higher than that in patients with oligoarthritis (12.7 and 11.0 vs. 4.2 pg/mL, both P<0.05). A positive correlation of IL-6 or TNF-α level with C reactive protein was observed in patients with ERA.

CONCLUSIONSThe results indicate that Th1/Th2 imbalance and Th1 predominance may exist in children with non-SOJIA; and IL-6 may be involved in the pathogenesis of non-SOJIA children with polyarthritis.

Mycobacterium (M.) vaccae is a fast-growing species of saprophytic bacteria that is widely distributed. To understand the host immune responses induced by M. vaccae isolated from bovine submaxillary lymph nodes, C57BL/6 mice were infected with reference strain M. vaccae Bacillus Calmette-Guérin (BCG) and isolated M. vaccae using intraperitoneal injections. Comparison of the bacterial replication and organ pathology between M. vaccae and M. vaccae BCG revealed that M. vaccae was more malignant than M. vaccae in mice. We also demonstrated that serum from the M. vaccae-infected mice contained a higher expression level of gamma-interferon (IFN-γ), tumor necrosis factor alpha, monocyte chemoattractant protein-1, interleukin (IL)-4, IL-12, IL-10 and transforming growth factor beta than did the other groups, especially after week 4. Furthermore, when the numbers of CD3⁺CD4⁺IFN-γ⁺ and CD3⁺CD4⁺IL4⁺ cells in the infected mice were observed by flow cytometry, we found that a powerful T helper 1 (Th1) response was induced by M. vaccae infection, which was associated with the emergence of CD3⁺CD4⁺IFN-γ⁺ cells. However, the Th1 response declined over time, which was associated with appearance of the CD4⁺CD25⁺FoxP3⁺ and CD4⁺CD25⁺CD152⁺Treg cell reaction. In addition, a strong Th2 response was found. Finally, we found that M. vaccae infection increased the production of type I IFNs, which was associated with a reduced Th1 response.
Animals ; Bacillus ; Bacteria ; Chemokine CCL2 ; Flow Cytometry ; Injections, Intraperitoneal ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukins ; Lymph Nodes ; Mice* ; Mycobacterium bovis ; Mycobacterium* ; Pathology ; Transforming Growth Factor beta ; Tumor Necrosis Factor-alpha

OBJECTIVE:To establish the ion chromatography for the mass fractions of 9 inorganic anions (fluoride ion,for-mate radical,chloride ion,nitrite radical,bromide ion,nitrate radical,sulfate radical,oxalate radical and phosphate radical) in Ganmao qingre granules. METHODS:Ion chromatograph was used. The column was Ion Pac AS11-HC with the mobile phase of po-tassium hydroxide solution by gradient elution(concentration:30 mmol/L,time:15 min),the column temperature was room tem-perature,the volume was 25 μl. RESULTS:The 9 inorganic anions were basically achieved separation and had good linear relation-ship with r of 0.999 2-0.999 9,respectively;RSDs of precision,stability and reproducibility tests were less than 3.50%. Average re-covery was 92.47%-103.51%(RSD=0.42%-3.41%,n=6),respectively. CONCLUSIONS:The established method is fast,accu-rate,sensitive and repeatable and can be used for the determination of 9 inorganic anions in Ganmao qingre granules.