ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

Objective:To explore potential predictors of refractory Mycoplasma pneumoniae pneumonia (RMPP) in early stage. Methods:The prospective multicenter study was conducted in Zhejiang, China from May 1 st, 2019 to January 31 st, 2020. A total of 1 428 patients with fever >48 hours to <120 hours were studied. Their clinical data and oral pharyngeal swab samples were collected; Mycoplasma pneumoniae DNA in pharyngeal swab specimens was detected. Patients with positive Mycoplasma pneumoniae DNA results underwent a series of tests, including chest X-ray, complete blood count, C-reactive protein, lactate dehydrogenase (LDH), and procalcitonin. According to the occurrence of RMPP, the patients were divided into two groups, RMPP group and general Mycoplasma pneumoniae pneumonia (GMPP) group. Measurement data between the 2 groups were compared using Mann-Whitney U test. Logistic regression analyses were used to examine the associations between clinical data and RMPP. Receiver operating characteristic (ROC) curves were used to analyse the power of the markers for predicting RMPP. Results:A total of 1 428 patients finished the study, with 801 boys and 627 girls, aged 4.3 (2.7, 6.3) years. Mycoplasma pneumoniae DNA was positive in 534 cases (37.4%), of whom 446 cases (83.5%) were diagnosed with Mycoplasma pneumoniae pneumonia, including 251 boys and 195 girls, aged 5.2 (3.3, 6.9) years. Macrolides-resistant variation was positive in 410 cases (91.9%). Fifty-five cases were with RMPP, 391 cases with GMPP. The peak body temperature before the first visit and LDH levels in RMPP patients were higher than that in GMPP patients (39.6 (39.1, 40.0) vs. 39.2 (38.9, 39.7) ℃, 333 (279, 392) vs. 311 (259, 359) U/L, both P<0.05). Logistic regression showed the prediction probability π=exp (-29.7+0.667×Peak body temperature (℃)+0.004×LDH (U/L))/(1+exp (-29.7+0.667×Peak body temperature (℃)+0.004 × LDH (U/L))), the cut-off value to predict RMPP was 0.12, with a consensus of probability forecast of 0.89, sensitivity of 0.89, and specificity of 0.67; and the area under ROC curve was 0.682 (95% CI 0.593-0.771, P<0.01). Conclusion:In MPP patients with fever over 48 to <120 hours, a prediction probability π of RMPP can be calculated based on the peak body temperature and LDH level before the first visit, which can facilitate early identification of RMPP.

Previous research on traditional Chinese medicine tongue inspection has mainly focused on its development,clinical application,and objectification,ignoring the fundamental question of "why tongue inspection is important in traditional Chinese medicine diagnosis" and neglecting the dominant role of traditional Chinese medicine brain thinking activities during diagnosis. Therefore,applying modern cognitive neuroscience technology to explore the impact of tongue inspection information on traditional Chinese medicine diagnostic thinking from an empirical research perspective may open up a new path for future tongue inspection research. This article proposes the combined application of eye tracking and event-related potential technology,drawing on the experimental paradigm of text reading inference correction in cognitive psychology,to collect eye movement and electroencephalogram data of participants induced by changes in tongue information during the reading process of traditional Chinese medicine medical records. This method can achieve synchronous observation of the focus areas and electroencephalogram signals of the subjects during the reading process,and provide cognitive scientific evidence for "why tongue inspection is important in traditional Chinese medicine diagnosis" through statistical data analysis. This research approach integrates the theories,techniques,and methods of traditional Chinese medicine and cognitive science,and is of great significance for developing cognitive thinking research in traditional Chinese medicine.

Objective:To retrospectively analyze the independent risk factors of complicated appendicitis(CA)in children under five years old and establish a clinical prediction model, and to evaluate the clinical application of this model.Methods:A retrospective analysis was performed on children under five years old who underwent appendectomy at Children′s Hospital of Shanghai Jiao Tong University School of Medicine from January 2018 to December 2021.The children were divided into CA group and uncomplicated appendicitis group according to whether there was sign of perforation or gangrene in appendiceal tissue after operation.The differences in clinical features and preoperative laboratory test results between two groups were compared.The independent risk factors of CA were identified and a clinical prediction model was established.The clinical prediction model was verified by receiver operating characteristic curve.Results:A total of 140 children were enrolled in this study, including 84 cases in the CA group and 56 cases in uncomplicated appendicitis group.Univariate and binary Logistic regression analysis showed that the duration of symptoms>23.5 h( OR=6.650, 95% CI 2.469-17.912, P<0.05), abdominal muscle tension( OR=3.082, 95% CI 1.190-7.979, P<0.05) and C-reactive protein>41 mg/L ( OR=3.287, 95% CI 1.274-8.480, P<0.05) were independent risk factors for CA( P<0.05). The clinical prediction model of CA was constructed by the above mentioned three independent risk factors.The area under the receiver operating characteristic curve of the clinical prediction model was 0.881(95% CI 0.825-0.936), the sensitivity was 77.4%, the specificity was 87.5%, the positive predictive value was 91.3% and the negative predictive value was 70.0%. Conclusion:Acute appendicitis in children under five years old is more likely to progress to CA if the duration of symptoms>23.5 h, the level of C-reactive protein is increased, and the abdominal muscle tension is accompanied.The clinical prediction model of CA constructed by common clinical information in pediatric clinics has good prediction efficiency, which provides a simple and feasible reference method for clinicians to distinguish CA from uncomplicated appendicitis.

Objective:To analyse China′s Traditional Chinese Medicine(TCM) health service policy from the perspective of policy life cycle, so as to provide reference for improving the policy system of TCM health service.Methods:The official websites of national government agencies were searched for policy documents on TCM health service from 2012 to 2022, using the keywords of " Chinese medicine" " Traditional Chinese Medicine" and " health service", and policy text entries were extracted and coded. A two-dimensional analytical framework of policy tools(supply, demand, and environment) and policy life cycle(agenda setting, policy formulation, policy implementation, and policy evaluation) was constructed to quantitatively analyze the policy texts.Results:A total of 58 policy documents were included, containing 2 898 text entries. The distribution of policy instruments was uneven, with a high percentage of entries using supply-based instruments(40.57%, 1 176 entries), followed by environmental instruments(38.86%, 1 126 entries) and demand-based instruments(20.57%, 596 entries). From the perspective of policy cycle, the policy tools were mainly concentrated in the stages of policy formulation(53.77%, 1 558 entries) and policy implementation(36.86%, 1 068 entries), while the policy tools in the stages of agenda setting(4.97%, 144 entries) and policy evaluation(4.40%, 128 entries) needed to be strengthened.Conclusions:In recent years, the distribution of policy tools in China′s TCM health service policy was uneven, and the articulation of the stages in the policy life cycle needed to be strengthened.

Objective:To investigate how medical students process Western medicine(WM)terms vs.traditional Chinese medicine(TCM)terms and how this is impacted by analogical priming from the perspective of cognitive neuroscience.Methods:The experiments were designed as a 2(TCM terms vs.WM terms)x 2(correct terms vs.incorrect terms)x 2(analogical priming task vs.non-priming task)scheme.A total of 26 medical stu-dents completed a non-priming judgment task and an analogical priming judgment task on medical terms.During the tasks,the participants were asked to make correct/incorrect judgments on WM terms and TCM terms,and their behavioral data and event-related potentials(ERPs)were recorded.Results:Behaviorally,the response speed and accuracy of WM terms were higher than those of TCM terms(both P＜.001),indicating a prominent concreteness effect.Analogical priming shortened the response time to medical terms(P＜.001),and the response time to TCM terms was shortened more significantly(P=.001).For ERPs,WM terms induced a larger P200(an early positive ERP component),a smaller N400(a negative ERP component),and a higher late positive ERP component,indicating supe-riority of attention adjustment,smaller-scale semantic activation,and a higher cost of late semantic analysis and integration.However,the analogical priming eliminated the difference between WM and TCM terms in P200 and N400 while maintaining it in the LPC.This suggests that WM terms are sensitive to analogical attention adjustment,and TCM terms are susceptible to analogical semantic integration.Conclusion:Our findings suggest that WM and TCM disease behavior terms may initially differ in concreteness or the use of a verbal-linguistic system.Analogy is more conducive to understanding TCM terms.This research provides important neuroscientific evidence of the difference in thinking between TCM and WM.

Objective:To evaluate the clinical efficacy of extensively hydrolyzed protein formula(eHF) in very low birth weight(VLBW) infants.Methods:A prospective controlled signal-center trial was conducted in this study, the preterm infants with gestational age of 28-33 weeks and birth weight of 999-1 500 g who were hospitalized at Department of Neonatology, Zhuhai Maternal Hospital within the first 12 hours between January and December 2018, were selected.They were assigned into breast feeding group(HM) and formula feeding group according to the mothers′ disease and parents′ breastfeeding willingness.The formula feeding group was assigned into eHF group and preterm formula (PF)group according to the parents′ breastfeeding willingness.The infants discharging or dead before achieving full feeding, discharging within 28 days after birth, with congenital malformation (complex congenital heart disease, digestive system malformation, etc) and severe sepsis were rolled out. Chi- square test and One- Way ANOVA were used for statistical analysis.Prospective study was conducted among the 3 groups comparing the incidence of feeding intolerance, duration of meconium discharge, the time to regain birth weight and reach full enteral feeding, average hospital stay, incidence of necrotizing enterocolitis of newborn(NEC), cholestasis and extrauterine growth retardation(EUGR), the growth rate of head circumference, length and weight in the first 4 weeks of life, and blood biochemical indices at the first 2 weeks and 4 weeks of life. Results:A total of 102 infants were enrolled, 35 cases in the eHF group, 37 cases in the PF group and 30 cases in the HM group.Compared with the PF group [54.0%(20/37 cases)], the eHF group[22.0%(8/35 cases)] and the HM group [16.7%(5/30 cases)] had lower incidence of feeding intolerance, and the differences were statistically significant( χ2=7.366, 9.901, all P<0.05). The time to regain birth weight[(8.9±1.8) d, (9.1±1.4) d vs.(10.8±2.9) d], time for achieving full enteral feeding [(42.8±2.8) d, (42.3±3.3) vs.(45.5±3.4) d], the duration of meconium discharge [(7.2±1.8) d, (6.6±1.8) d vs.(8.7±2.1) d], and average hospital stay [(52.9±1.1) d, (52.3±1.2) d vs.(54.1±1.2) d]in the eHF group and HM group were shorter than those in PF group, and the differences were statistically significant(all P<0.05); and there was no statistically significant difference between the eHF group and the HM group(all P>0.05). There was no statistically significant difference in the incidence of incidence of NEC, cholestasis and EUGR, the growth rate of head circumfe-rence, length and weight in the first 4 weeks of life, the serum albumin, creatinine, urea nitrogen at first 2 weeks and 4 weeks of life among the 3 groups (all P>0.05). Compared with the PF group, the serum total bilirubin levels in the eHF group and the HM group were lower at 2 weeks [(109.4± 4.6) μmol/L, (110.2±1.0) μmol/L vs.(115.0±7.6) μmol/L]and 4 weeks after birth[(79.3±9.7) μmol/L, (80.0±1.7) μmol/L vs.(81.5±8.4)μmol/L], and the differences were statistically significant(all P<0.05), but no statistically significant difference was found between the eHF goup and the HM group(all P>0.05). Conclusions:For VLBW infants, eHF can reduce feeding intole-rance, promote defecation, achieve full feeding faster, promote bilirubin metabolism, shorten hospital stay, does not affect growth and development in short-term.

A case of ichthyosis follicularis,alopecia and photophobia syndrome caused by a novel mutation c.1165C＞T in the membrane-bound transcription factor protease site 2 (MBTPS2) gene was firstly reported.The proband presented with dry skin,congenital hairlessness,follicular keratotic papules,photophobia,epilepsy,and mental and motor retardation.Next-generation and Sanger sequencing analysis confirmed that the proband and his mother both had a c.1165C＞T (p.pro389Ser) mutation in exon 9 of the MBTPS2 gene.According to the clinical manifestations of the patient and genetic characteristics of the MBTPS2 gene mutation,the patient was diagnosed with ichthyosis follicularis,alopecia and photophobia syndrome.

Clinical features of and genetic mutations in two cases of pseudohypoparathyroidism type Ⅰ a(PHP Ⅰ a) with early-onset skin nodules were analyzed.Both of the two patients were males,and their ages at onset were 2 and 3 months respectively.They both presented with early-onset skin nodules as the main clinical manifestation,and were clinically characterized by a round face,short neck and early obesity.Histopathological examination of skin lesions showed subcutaneous ectopic osteogenesis in both patients.The first patient had low blood calcium,high blood phosphorus,high parathyroid hormone (PTH),and gene sequencing showed a heterozygous mutation c.399delT causing a T base deletion at position 399 in exon 5 of the GNAS gene.The second patient had normal blood calcium and phosphorus levels as well as normal PTH levels at early stage,and gene sequencing showed a heterozygous mutation c.939delT causing a T base deletion at position 939 in exon 9 of the GNAS gene.The blood PTH level was found to increase in the second patient after 1-year follow-up.Both the patients were confirmedly diagnosed with PHP Ⅰa.After treatment with vitamin D3,no new skin nodules occurred,and the blood calcium and phosphorus levels returned to normal.