Objective To investigate the protective effect of herba artemisiae scopariae extract on multidrug resistance protein 3(MDR3)gene mutation-induced neonatal parenteral nutrition-associated cholestasis(PNAC)and its possible mechanism.Methods ①Human primary hepatocytes were treated with cell culture in vitro,CRISPR/Cas9 lentivirus infection and MDR3 mutant gene lead-in.The levels of hepatic and biliary biochemical indexes[alanine transaminase(ALT),aspartate transaminase(AST),total bilirubin(TBil),direct bilirubin(DBil),indirect bilirubin(IBil),total bile acid(TBA)]in the supernatant of hepatocytes before and after 16,32,48 hours were compared to determine the time required for fatty acid induction of PNAC hepatocyte model with MDR3 gene mutation.② Human primary hepatocytes were divided into blank control group,MDR3 gene wild type group,MDR3 gene mutation group,and herba artemisiae scopariae extract intervention group according to random number table method.The blank control group was treated with culture medium only,the MDR3 gene wild type group was infected with lentivirus and mixed with wild type MDR3 gene and culture medium,the MDR3 gene mutation group was infected with lentivirus and cultured in culture medium with the mutant genes lead-in of LV-MDR3KI(c.485T>A,c.2793insA,c.1031G>A,c.3347G>A)mutation,while the MDR3 mutant gene was lead-in by lentivirus infection and cultured in culture medium,and then pretreated with 100 g/L herba artemisiae scopariae extract in the herba artemisiae scopariae extract intervention group,then the four groups of hepatocytes were induced with 1%fat emulsion,and the treatment time was the time needed to construct the PNAC hepatocytes model with MDR3 gene mutation.The levels of ALT,AST,TBil,DBil,IBil and TBA in the supernatant of hepatocytes were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression abundance of adenosine triphosphate binding cassette proteins(ABCB4,ABCB11,ABCC2,ABCC3,ABCC4)encoding MDR3,bile salt export pump(BSEP),multidrug resistance associated protein(MRP)2-4,and tumor necrosis factor-α(TNF-α)genes were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared to the blank control group and MDR3 gene wild type group,there was no significant difference in the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 gene mutant group before and 16 hours after induction with 1%fat emulsion,however after treated with 1%fat emulsion for 32 hours and 48 hours,the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 mutant hepatocytes were significantly increased(P<0.05),consequently the time required for fatty acid induction of PNAC hepatocyte model was 32 hours.At 32 hours after treatment with fat emulsion,the levels of ALT,AST,TBil,DBil,TBA in the supernatant of hepatocytes in the herba artemisiae scopariae extract intervention group were significantly decreased[ALT(ng/L):148.3±2.3 vs.164.9±7.0,AST(ng/L):2767.4±78.8 vs.3239.4±107.1,TBil(μmol/L):7.6±0.2 vs.13.6±0.3,DBil(μmol/L):1.8±0.1 vs.5.7±0.2,TBA(μmol/L):3.4±0.2 vs.6.7±0.1,all P<0.05].The ABCB4,ABCC2,ABCC3,ABCC4 mRNA expression of MDR3,MRP2,MRP3,MRP4 in the blank control group,MDR3 wild type group,MDR3 gene mutation group and the herba artemisiae scopariae extract intervention group had no significant difference.The expression of TNF gene mRNA was highly expressed in MDR3 gene mutation group(2-??Ct:1.258±0.200 vs.1.001±0.052),and was low expressed in the herba artemisiae scopariae extract intervention group(2-??Ct:0.387±0.247 vs.1.258±0.200),and there was a significant difference between the two groups(both P<0.05).Compared to the MDR3 gene mutation group,the ABCB11 gene encoding BSEP mRNA expression in the herba artemisiae scopariae extract intervention group was significantly increased(2-??Ct:2.955±0.479 vs.1.333±0.529,P<0.05).Conclusion The herba artemisiae scopariae extract has a protective effect on PNAC induced by MDR3 gene mutation,which may be related to antagonizing inflammatory reaction,decreasing the expression of TNF mRNA and improving the expression of ABCB11 gene encoding BSEP.

Objective:In order to explore the impact of corona virus disease 2019(COVID-19)on the hospitalization of children with bronchiolitis and to improve clinicians′ understanding of the characteristics of bronchiolitis during the COVID-19 epidemic.Methods:This was a multicenter clinical study, and the data have been collected from 23 children′s medical centers in China.All the clinical data were retrospectively collected from children with bronchiolitis who were hospitalized at each study center from January 1, 2019 to December 31, 2021.The results included gender, age at hospitalization, length of stay, respiratory syncytial virus(RSV) test results, severity rating, ICU treatment, and the total number of children hospitalized with respiratory tract infection during the same period.The clinical data of children with bronchiolitis in 2019 before COVID-19 epidemic and in 2020、2021 during COVID-19 epidemic were statistically analyzed and compared.Results:According to a summary of data provided by 23 children′s medical centers, there were 4 909 cases of bronchiolitis in 2019, 2 654 cases in 2020, and 3 500 cases in 2021.Compared with 2019, the number of bronchiolitis cases decreased by 45.94% in 2020 and 28.70% in 2021.In 2019, 2020 and 2021, there were no significant differences in gender ratio, age, and duration of hospitalization.Compared with 2019, the ratio of bronchiolitis to the total number of hospitalizations for respiratory tract infection decreased significantly in 2020 and 2021( χ2=12.762, P<0.05; χ2=84.845, P<0.05).The proportion of moderate to severe bronchiolitis cases in both 2020 and 2021 was lower than that in 2019, and the difference was statistically significant ( χ2=4.054, P<0.05; χ2=8.109, P<0.05).There was no statistically significant difference in the proportion of bronchiolitis cases requiring ICU treatment between 2019, 2020, and 2021 ( χ2=1.914, P>0.05).In 2019, a total of 52.60%(2 582/4 909) of children with bronchiolitis underwent RSV pathogen testing, and among them, there were 708 cases with RSV positive, accounting for 28.00%.In 2020, 54.14%(1 437/2 654) of children with bronchiolitis underwent RSV pathogen testing, and there were 403 cases with RSV positive, accounting for 28.04%.In 2021, 66.80%(2 238/3 500) of children with bronchiolitis underwent RSV pathogen testing, and there were 935 cases with RSV positive, accounting for 41.78%.Compared with 2019 and 2020, the RSV positive rate in 2021 showed a significant increase( χ2=99.673, P<0.05; χ2=71.292, P<0.05). Conclusion:During the COVID-19 epidemic, the implementation of epidemic prevention and control measures reduced the hospitalization rate and severity of bronchiolitis, but did not reduce the positive rate of RSV detection.

Objective:To investigate the clinical significance of detecting serum 25-hydroxyl-vitamin D [25(OH)D] level and albumin/fibrinogen ratio (AFR) in patients with rheumatoid arthritis (RA).Methods:This retrospective study included 131 patients (95 patients with RA and 36 with arthralgia excluding autoimmune diseases), who were treated in Bozhou People′s Hospital from May 2017 to January 2020. Forty healthy controls underwent health checkups during the same period served as healthy control. RA group was divided into high (>5.1, 25 cases), medium (3.2

Objective:To investigate the knowledge, attitudes, and practices (KAP) of pulse oximetry among pediatric healthcare providers in China and analyze the factor influencing the KAP.Methods:A self-developed questionnaire was used for an online research on the KAP of 11 849 pediatric healthcare providers from 31 provinces, autonomous regions, and municipalities of China from March 11 to 14, 2022.The factors influencing the KAP of pulse oximetry among pediatric healthcare providers were examined by Logistic regression. Results:The scores of KAP, of pulse oximetry were 5.57±0.96, 11.24±1.25 and 11.19±4.54, respectively.The corresponding scoring rates were 69.61%, 74.95%, and 55.99%, respectively. Logistic regression results showed that the gender and working years of pediatric healthcare providers, the region they were located, and whether their medical institution was equipped with pulse oximeters were the main factors affecting the knowledge score (all P<0.05). Main factors influencing the attitude score of pediatric healthcare providers included their knowledge score, gender, educational background, working years, region, medical institution level, and whether the medical institution was equipped with pulse oximeters (all P<0.05). For the practice score, the main influencing factors were the knowledge score, gender, age, and whether the medi-cal institution was equipped with pulse oximeters (all P<0.05). Conclusions:Chinese pediatric healthcare providers need to further improve their knowledge about and attitudes towards pulse oximetry.Pulse oximeters are evidently under-used.It is urgent to formulate policies or guidelines, strengthen education and training, improve knowledge and attitudes, equip more institutions with pulse oximeters, and popularize their application in medical institutions.

AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.

Objective To assess the clinical diagnostic performance of liver stiffness measurement (LSM) and aspartate transaminase (AST)-to-platelet (PLT) ratio index (APRI) for liver fibrosis in chronic hepatitis B (CHB) patients with alanine aminotransferase (ALT) less than or equal to five times of the upper limit of normal (≤5×upper limit of normal [ULN]).Methods FibroScan,blood routine and liver function test were conducted at the day or one day before liver biopsy in 383 CHB patients with ALT≤5 × ULN.The Scheuer scoring system was used for liver histologic assessment.APRI was calculated.Based on the results of liver pathology,the areas under receiver operating characteristic curve (AUC) of LSM and APRI for diagnosis of liver fibrosis stage were compared.Results The median LSM were 5.10 kPa for S0 fibrosis stage,5.20 kPa for S1,6.60 kPa for S2,10.10 kPa for S3,and 18.80 kPa for S4.The median APRI values were 0.36,0.38,0.63,0.61 and 1.27,respectively.The AUC of LSM were 0.817 for ≥S2,0.891 for ≥S3 and 0.913 for ≥S4.And the AUC of APRI were 0.717 for ≥S2,0.711 for ≥S3 and 0.746 for ≥S4.The cut-offs of LSM values were 6.8 kPa for ≥S2,8.7 kPa for ≥S3,and 10.9 kPa for ≥S4.Conclusion LSM can accurately assess the degree of liver fibrosis in CHB patients with ALT ≤5 × ULN,which is superior to APRI in clinical utility.

BACKGROUND:Ability of adipose mesenchymal stem cels differentiating into type II alveolar epithelial cels and the regulating mechanism have not been fuly elucidated.
OBJECTIVE:To study the ability of adipose mesenchymal stem cels differentiating into type II alveolar epithelial cels in vitro and the function of Wnt pathway in the regulation of differentiation.
METHODS:Adipose mesenchymal stem cels were obtained from fat tissue of rats and identified by flow cytometry. The adipose mesenchymal stem cels were divided into control group, smal airway growth medium (SAGM) group and Wnt3a group. Control group was treated with normal DMEM medium; SAGM and Wnt3a groups were both treated with smal airway growth medium, and additionaly, the Wnt3a group was treated with Wnt3a, a Wnt signaling pathway agonist. After 10 days, quantitative RT-PCR and immunofluorescence detection were used to test the expression of surfactant proteins B, C, D, type II alveolar epithelial markers. Phosphorylatedβ-catenin and GSK-3β were detected using western blot after 5 and 10 days of induction.
RESULTS AND CONCLUSION: Adipose mesenchymal stem cels with high purity could be successfuly isolated from the adipose tissue of rats, and expressed CD44 and CD29, but not CD11b and CD45. After SAGM treatment, protein and mRNA expressions of surfactant proteins B, C, D were al increased in adipose mesenchymal stem cels (P < 0.01), indicating the ability of adipose mesenchymal stem cels to be induced into type II epithelial cels. Surfactant proteins B, C, D expressions at protein and mRNA levels were significantly higher in the Wnt3a group than the SAGM group (P < 0.01). During the induction progress, the expression of phosphorylated β-catenin gradualy increased, but GSK-3βexpression gradualy decreased in the Wnt3a group (P < 0.01). These findings indicate that Wnt signaling pathways are involved in differentiation of adipose mesenchymal stem cels into type II alveolar epithelial cels.

Objective To analyze the factors related to the household abundance of rodents in rodent-borne disease foci in the western part of Yunnan province.Methods From July 2011 to October 2012,800 households (20 households in 1 village) were randomly selected in 40 natural villages of 10 counties in western Yunnan where rodent borne disease was endemic to conduct a study on relationship between rodent abundance and environmental factors.Five cages were placed in each household for 3 consecutive nights to capture rodents.The rodent species were identified based on their morphological characteristics.The data on potential factors related to rodent abundance were collected through questionnaires and field observation.A dataset was established by using EpiData software and the analysis was performed with hurdle regression model under R software.Results A total of 421 rodents were captured in 800 households,belonging to 9 species,6 genera,2 families,2 orders.Rattus tanezumi was the predominant species (66.03％).The final hurdle regression model showed that the probability of capturing rodents in the households where family member had high education level and the garbage was placed outside declined by 50％-68％ ; The probability of capturing rodents in the households of Dai and Yi ethnic groups increased by 2.16-2.87 times; The probability of capturing rodents in the households where rodents were observed or vegetables grown near houses increased by 1.54-1.59 times; In the households where many rodents were believed to exist,the probability of capturing rodents and the number of rodents captured increased by 1.59 and 1.84 times respectively.The number of rodents captured in the houses with cement or tile floor increased by 3.62 times.Conclusion The household abundance of rodents in the area in western Yunnan,where the rodent-borne disease survey was conducted,seemed to be closely related to the social economy status,human intervention and ecological environment.To control the abundance of rodents effectively,it is necessary to take these factors into consideration.

Objective:To investigate the effects of paeonol on aortic nuclear factor-kappaB ( NF-κB) activity and blood lipid levels in the rats with atherosclerosis ( AS) . Methods:The AS rat model was established by administration of high-fat diet and vitamin D3 ( i. p. ). The rats were divided into 5 groups (n=8) randomly, namely the normal group, model group, positive control group(simvasta-tin, 10 mg·kg-1 ) and paeonol groups with the dose of 20 and 10 mg·kg-1 , respectively. After the 4-week treatment, the serum of rats was collected to determine the lipid levels. The aortic pathologic changes of AS rats were observed under a microscope, and then graded, and aortic NF-κB activity was detected by an immunohistochemistry method. Results:Administration of paeonol improved the severity of aortic pathology in AS rats. The pathologic grade in the paeonol group (20 mg·kg-1 ) showed a significant difference com-pared with that in the model group (P<0. 05). Paeonol (20 and 10 mg·kg-1) markedly decreased the TG, TC and LDL-C levels in the serum of AS rats (P<0. 05 or P<0. 01). Meanwhile, paeonol significantly inhibited the aortic NF-κB activity(IOD value) with (1. 96 ± 0. 55) and (2. 50 ± 0. 80) in 20 and 10 mg·kg-1 group, respectively, which showed significant differences compared with that in the model group (P<0. 05 or P<0. 01). Conclusion:Paeonol shows certain therapeutic effect in AS rats, and the mechanisms may be related with the regulation of lipid mobilization and the inhibition of aortic NF-κB activity.