To establish the amplification-refractory mutation system quantitative real-time PCR (ARMS-qPCR) method based on qPCR technique for detecting the c.2T>C mutation of SLC25A13 gene and validate its diagnostic performance. According to the principle of ARMS-qPCR primer design, the specific primers were designed for the conserved sequence of SLC25A13. The c.2T>C mutation ARMS-qPCR detection assay of SLC25A13 gene and the corresponding Sanger sequencing system were established through the use of the synthetic plasmids of homozygous mutation and 200 human peripheral blood specimens which were verified by Sanger sequencing as templates, and the diagnostic efficacy of the qPCR assay was validated by using nucleic acid extracted from another 200 human peripheral blood specimens and the results obtained were compared with the Sanger sequencing results as the gold standard, and the consistency of the two detection methods was analyzed. The results showed that the qPCR assay could accurately identify artificial plasmids carrying different mutations of SLC25A13 gene, and distinguish between wild type SLC25A13 gene and the c.2T>C mutation. This method was used to detect the mutation status of SLC25A13 c.2T>C in human peripheral blood, and the detection results were 100% consistent with the Sanger sequencing results. Among the 200 blood samples, 8 samples (4%) carried the c.2T>C mutation of SLC25A13 gene and 192 samples (96%) did not carry it. In conclusion, the ARMS-qPCR test established in this study can quickly, simply and accurately detect the c.2T>C mutation of SLC25A13 gene, which is helpful for the diagnosis of citrin deficiency (CD).

To establish the amplification-refractory mutation system quantitative real-time PCR (ARMS-qPCR) method based on qPCR technique for detecting the c.2T>C mutation of SLC25A13 gene and validate its diagnostic performance. According to the principle of ARMS-qPCR primer design, the specific primers were designed for the conserved sequence of SLC25A13. The c.2T>C mutation ARMS-qPCR detection assay of SLC25A13 gene and the corresponding Sanger sequencing system were established through the use of the synthetic plasmids of homozygous mutation and 200 human peripheral blood specimens which were verified by Sanger sequencing as templates, and the diagnostic efficacy of the qPCR assay was validated by using nucleic acid extracted from another 200 human peripheral blood specimens and the results obtained were compared with the Sanger sequencing results as the gold standard, and the consistency of the two detection methods was analyzed. The results showed that the qPCR assay could accurately identify artificial plasmids carrying different mutations of SLC25A13 gene, and distinguish between wild type SLC25A13 gene and the c.2T>C mutation. This method was used to detect the mutation status of SLC25A13 c.2T>C in human peripheral blood, and the detection results were 100% consistent with the Sanger sequencing results. Among the 200 blood samples, 8 samples (4%) carried the c.2T>C mutation of SLC25A13 gene and 192 samples (96%) did not carry it. In conclusion, the ARMS-qPCR test established in this study can quickly, simply and accurately detect the c.2T>C mutation of SLC25A13 gene, which is helpful for the diagnosis of citrin deficiency (CD).

Bupleuri radix is the dried root of Bupleuri radix or narrow-leaved Bupleuri radix of the umbelliferae family,and it is the most commonly used traditional Chinese medicine,which was first published in Shennong's Classic of Materia Medica.It has a variety of pharmacological effects such as anti-inflammatory,antioxidant,hepatoprotective,antitumor,antidepressant,et al.In the modern study,the extract of Bupleuri radix mainly includes a variety of chemical components such as Bupleuri radix saponin,flavonoids and volatile oil.By reviewing the relevant literature at home and abroad,this paper summarizes the research progress on the chemical composition and pharmacological effects of Bupleuri radix,and points out the future research direction to provide a certain reference basis for the subsequent research.

Purpose: This study aims to evaluate the efficacy and safety of a new combination treatment of vinorelbine and pyrotinib in human epidermal growth factor receptor 2 (HER2)–positive metastatic breast cancer (MBC) and provide higher level evidence for clinical practice. Materials and Methods: This was a prospective, single-arm, phase 2 trial conducted at three institutions in China. Patients with HER2-positive MBC, who had previously been treated with trastuzumab plus a taxane or trastuzumab plus pertuzumab combined with a chemotherapeutic agent, were enrolled between March 2020 and December 2021. All patients received pyrotinib 400 mg orally once daily plus vinorelbine 25 mg/m2 intravenously or 60-80 mg/m2 orally on day 1 and day 8 of 21-day cycle. The primary endpoint was progression-free survival (PFS), and the secondary endpoints included the objective response rate (ORR), disease control rate (DCR), overall survival, and safety. Results: A total of 39 patients were enrolled. All patients had been pretreated with trastuzumab and 23.1% (n=9) of them had accepted trastuzumab plus pertuzumab. The median follow-up time was 16.3 months (95% confidence interval [CI], 5.3 to 27.2), and the median PFS was 6.4 months (95% CI, 4.0 to 8.8). The ORR was 43.6% (95% CI, 27.8% to 60.4%) and the DCR was 84.6% (95% CI, 69.5% to 94.1%). The median PFS of patients with versus without prior pertuzumab treatment was 4.6 and 8.3 months (p=0.017). The most common grade 3/4 adverse events were diarrhea (28.2%), neutrophil count decreased (15.4%), white blood cell count decreased (7.7%), vomiting (5.1%), and anemia (2.6%). Conclusion Pyrotinib plus vinorelbine showed promising efficacy and tolerable toxicity as second-line treatment in patients with HER2-positive MBC.

Objective:To compare the differences in exploratory behaviors between infants with high-risk autism spectrum disorder (HR-ASD) and normal development (TD) infants, and to explore the relationship between exploratory behaviors and the severity of ASD symptoms.Methods:A total of 31 infants aged 6-23 months with HR-ASD in the Pediatric Clinic, the Affiliated Brain Hospital of Nanjing Medical University from January 2019 to August 2020 were retrospectively recruited in the HR-ASD group, and 37 TD infants aged from 6 to 23 months in Nanjing were selected as the TD group.The development level of the 2 groups was evaluated by Gesell development scale, and the depth and breadth of exploratory behaviors were measured in free play.Autism Diagnostic Observation Schedule (ADOS) was used to evaluate the severity of ASD symptoms in HR-ASD group.Differences in exploratory behaviors between the 2 groups were analyzed by the independent sample t-test and Chi- square test. Pearson′ s correlation analysis was performed to explore the correlation between exploratory behaviors and symptom severity in HR-ASD group.Binary Logistic regression analysis was used to explore the discrimination efficiency of sexual behavior in the 2 groups. Results:Compared with TD group, the depth and breadth of exploratory behaviors in HR-ASD group were significantly lower (55.06±25.73 vs.132.78±44.69, 4.42±2.20 vs.8.78±3.28)( t=-8.95, -6.53, all P<0.01), and atypical exploratory behavior and avoidance withdrawal behavior were significantly worse [18 cases(58.06%) vs.6 cases(16.22%), 10 cases(32.26%) vs.3 cases(8.11%)]( χ2=15.30, 6.36, all P<0.05). Significant differences in the frequency and duration of object exploration, environmental exploration and social exploration were detected between HR-ASD group and TD group (3 vs. 0 vs. 0, 45 vs. 0 vs. 0)(all P<0.01). Pearson correlation analysis showed that, the frequency of object exploratory behavior was positively correlated with the dimension of ADOS in HR-AD group ( r=0.40, P<0.05); while the duration of object exploratory behavior, the complexity of object explo-ratory behavior and the depth of exploratory behavior were negatively correlated with CARS score ( r=-0.45, -0.47, -0.42, all P<0.05). The depth of exploratory behavior was negatively correlated with the stereotyped dimension of ADOS ( r=-0.40, P<0.05). Binary Logistic regression analysis showed that the depth of exploratory behavior had a discrimination effect ( P<0.01). Conclusions:The characteristics of exploratory behaviors in HR-ASD infants are abnormal, especially the distinguishing effect on the depth of exploratory behaviors, which can be used as a candidate behavioral index for early screening of ASD.

Objective:To explore the significance of empathy and executive function indexes in the diagnosis and classification of autism spectrum disorder (ASD) and attention deficit hyperactivity disorder (ADHD).Methods:According to DSM-Ⅴ diagnostic criteria, 33 children with ASD, 30 children with ADHD and 39 typical development (TD) children and adolescents were enrolled as the research subjects.The empathy and executive function characteristics were compared and analyzed in the subjects.Based on empathy and executive function indicators, the three groups of subjects were diagnosed and classified by machine learning method.Results:The total score of Griffith empathy measure parent ratings(GEM-PR)(ASD: (0.67±0.64), ADHD: (1.00±0.79), TD: (0.98±0.73)) and each factor score ( F=3.595-10.363, all P<0.05) and the total score of behavior rating inventory of executive function(BRIEF)(ASD: (62.79±7.45), ADHD: (59.47±8.77), TD: (49.08±7.91)) and each factor score of the three groups were different ( F=6.557-33.205, all P<0.01). Among them, the scores of empathy and executive function in ASD and ADHD groups were generally higher than those in TD children (all P<0.05). When combined with BRIEF scale and GEM scale, the classification accuracy of the three groups reached 62.75%. Conclusion:Both ASD and ADHD children have damages in empathy and executive function.Combining empathy and executive function indexes are more helpful for diagnosis and classification than single index.

Objective:To evaluate the cost effectiveness of primary prophylaxis (PP) with pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF), PP with recombinant human granulocyte colony stimulating factor (rhG-CSF) and no prophylaxis in women with early-stage breast cancer in China.Methods:Two phase Markov models were constructed for a hypothetical cohort of patients aged 45 with stage Ⅱ breast cancer. The first phase modelled costs and outcomes of 4 cycles docetaxel combined with cyclophosphamide [TC×4, febrile neutropenia (FN) risk>20%] chemotherapy, which assumptions based on literature reviews, including FN rates [base-case (deterministic sensitivity analysis range), 0.29 (0.24-0.35)] and related events [FN case-fatality, 3.4 (2.7-4.1)]. Second phase modelled the long term survival which was link with the relative dose intensity (RDI) [mortality hazard ratio ( HR) of RDI < 85% vs ≥85%, 1.45 (1.00-2.32)]. Clinical effectiveness, therapeutic costs, and economic utilities were estimated from peer-reviewed publications and expert opinions in case of unavailability of published evidences. Results:Compared to rhG-CSF PP and no prophylaxis, the cost of PEG-rhG-CSF PP increased to 5 208.19 RMB and 5 222.73 RMB, respectively. The quality-adjusted life-years (QALYs) enhanced to 0.066 and 0.297, respectively. Accordingly, the incremental cost effectiveness ratios (ICERs) are 79 146.3 RMB and 17 558.77 RMB per QALY, which were both below the willingness to pay (WTP) threshold of three times GDP per capita (18, 000 RMB) recommended by the WHO. Sensitivity analysis suggested that the more clinically effective the primary prophylaxis with PEG-rhG-CSF is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. And the lower the mortality HR of RDI<85% vs ≥85% is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. Conclusion:Although the cost of PP PEG-rhG-CSF is higher, considering the additional benefits, the administrating of PP PEG-rhG-CSF is likely to be a cost-effective alternative to PP rhG-CSF and no prophylaxis in patients with early stage breast cancer whose FN risks are more than 20% in China.

Objective:To evaluate the cost effectiveness of primary prophylaxis (PP) with pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF), PP with recombinant human granulocyte colony stimulating factor (rhG-CSF) and no prophylaxis in women with early-stage breast cancer in China.Methods:Two phase Markov models were constructed for a hypothetical cohort of patients aged 45 with stage Ⅱ breast cancer. The first phase modelled costs and outcomes of 4 cycles docetaxel combined with cyclophosphamide [TC×4, febrile neutropenia (FN) risk>20%] chemotherapy, which assumptions based on literature reviews, including FN rates [base-case (deterministic sensitivity analysis range), 0.29 (0.24-0.35)] and related events [FN case-fatality, 3.4 (2.7-4.1)]. Second phase modelled the long term survival which was link with the relative dose intensity (RDI) [mortality hazard ratio ( HR) of RDI < 85% vs ≥85%, 1.45 (1.00-2.32)]. Clinical effectiveness, therapeutic costs, and economic utilities were estimated from peer-reviewed publications and expert opinions in case of unavailability of published evidences. Results:Compared to rhG-CSF PP and no prophylaxis, the cost of PEG-rhG-CSF PP increased to 5 208.19 RMB and 5 222.73 RMB, respectively. The quality-adjusted life-years (QALYs) enhanced to 0.066 and 0.297, respectively. Accordingly, the incremental cost effectiveness ratios (ICERs) are 79 146.3 RMB and 17 558.77 RMB per QALY, which were both below the willingness to pay (WTP) threshold of three times GDP per capita (18, 000 RMB) recommended by the WHO. Sensitivity analysis suggested that the more clinically effective the primary prophylaxis with PEG-rhG-CSF is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. And the lower the mortality HR of RDI<85% vs ≥85% is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. Conclusion:Although the cost of PP PEG-rhG-CSF is higher, considering the additional benefits, the administrating of PP PEG-rhG-CSF is likely to be a cost-effective alternative to PP rhG-CSF and no prophylaxis in patients with early stage breast cancer whose FN risks are more than 20% in China.

Objective To investigate the correlation between the feature of lobulation, degree of lobulation on CT images and epidermal growth factor receptor gene (EGFR) mutations in advanced pulmonary adenocarcinoma. Methods Two hundred and one patients who were diagnosed with advanced pulmonary adenocarcinoma between January and December 2017 and had undergone a chest CT and EGFR mutation testing at Shanghai Pulmonary Hospital were enrolled in this retrospective study. The clinical and imaging data of the patients were analyzed. The patients were classified into EGFR mutations group (107 cases) and wild type group (94 cases) according to whether EGFR gene mutation occurred. The feature of lobulation (described as presence of lobulation, absence of lobulation, or obscured margin), degree of lobulation (including deep lobulation, shallow lobulation, no lobulation, and obscured) on CT images were statistically compared between the two groups. Results Based on the presence or absence of lobulation sign, there were 99, 0, 8 cases with lobulation, no lobulation, obscured margin in the EGFR mutations group and 80, 1, 13 cases in the wild type group, respectively. There was no significant difference in lobulation sign between the EGFR mutations group and wild type group (P=0.152). According to the degree of lobulation, there were 32, 67, 0, 8 cases of deep lobulation, shallow lobulation, no lobulation, obscured margin in EGFR mutations group and 60, 20, 1, 13 cases in wild type group. Significant differences of frequency were found regarding deep lobulation and shallow lobulation between the two groups (P<0.001). However, there was no significant difference between the two groups in the presence of no lobulation, and obscured margin (P>0.05). Conclusions EGFR mutations are significantly associated with shallow lobulation in advanced pulmonary adenocarcinoma. Conversely, deep lobulation is more likely to appear in advanced pulmonary adenocarcinoma with wild?type EGFR. However, there is no association between EGFR mutation status and the presence of lobulation.

Objective: To investigate the correlation between the feature of lobulation, degree of lobulation on CT images and epidermal growth factor receptor gene (EGFR) mutations in advanced pulmonary adenocarcinoma. Methods: Two hundred and one patients who were diagnosed with advanced pulmonary adenocarcinoma between January and December 2017 and had undergone a chest CT and EGFR mutation testing at Shanghai Pulmonary Hospital were enrolled in this retrospective study. The clinical and imaging data of the patients were analyzed. The patients were classified into EGFR mutations group (107 cases) and wild type group (94 cases) according to whether EGFR gene mutation occurred. The feature of lobulation (described as presence of lobulation, absence of lobulation, or obscured margin), degree of lobulation (including deep lobulation, shallow lobulation, no lobulation, and obscured) on CT images were statistically compared between the two groups. Results: Based on the presence or absence of lobulation sign, there were 99, 0, 8 cases with lobulation, no lobulation, obscured margin in the EGFR mutations group and 80, 1, 13 cases in the wild type group, respectively. There was no significant difference in lobulation sign between the EGFR mutations group and wild type group (P=0.152). According to the degree of lobulation, there were 32, 67, 0, 8 cases of deep lobulation, shallow lobulation, no lobulation, obscured margin in EGFR mutations group and 60, 20, 1, 13 cases in wild type group. Significant differences of frequency were found regarding deep lobulation and shallow lobulation between the two groups (P<0.001). However, there was no significant difference between the two groups in the presence of no lobulation, and obscured margin (P>0.05). Conclusions EGFR mutations are significantly associated with shallow lobulation in advanced pulmonary adenocarcinoma. Conversely, deep lobulation is more likely to appear in advanced pulmonary adenocarcinoma with wild-type EGFR. However, there is no association between EGFR mutation status and the presence of lobulation.