Abstract: In the present study, the compound XL-12 from our previous work was utilized as a lead compound. Through the optimization of the terminal phenyl ring, 12 target compounds were designed and synthesized. The structures of all target compounds were confirmed by 1H NMR, 13C NMR, and H RMS. In vitro enzyme activity assay showed that most compounds demonstrated significant inhibitory activity toward Bruton’s tyrosine kinase (BTK) and Janus kinase 3 (JAK3). Among them, compound I-3 exhibited moderate cell proliferation inhibitory activity toward Daudi cells and BaF3-JAK3 cells. In the evaluation of anti-inflammatory activity in vitro, compound I-3 could effectively inhibit the production of inflammatory factors IL-6; besides, it exhibited superior anti-inflammatory activity compared to ibrutinib in xylene-induced ear swelling model in mice.

Objective To prepare and identify rabbit anti-cyclin dependent kinase 6 (CDK6) antibody. Methods The recombinant pET21a (+)/CDK6 was successfully constructed, then the recombinant plasmid was transformed into E.coli BL21 (DE3) competent cells and was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) for protein expression, which was detected by SDS-PAGE and Western blot analysis. The expressed protein was purified by nickel-chelating nitrilotriacetic acid (Ni-NTA) agarose and then analyzed by SDS-PAGE. Japanese white rabbits were immunized with purified CDK6 protein for many times every two weeks. The blood was collected at 0, 2, 4 and 6 weeks after immunization, and serum was separated from blood. The titer was detected by indirect ELISA. Western blot analysis, immunofluorescence assay and immunohistochemistry were employed to determine the specificity. Results High purity CDK6 protein and high specificity of rabbit anti-CDK6 antibody were successfully prepared. The titer of CDK6 rabbit serum antibody reached 1:30 000 after immunization, which could specifically recognize the CDK6 protein expressed in cervical cancer cell line and cervical cancer tissues. Conclusion The high titer and specificity of rabbit anti-CDK6 antibody is successfully prepared.
Animals ; Female ; Humans ; Rabbits ; Antibodies ; Antibody Specificity ; Blotting, Western ; Cyclin-Dependent Kinase 6 ; Enzyme-Linked Immunosorbent Assay ; Uterine Cervical Neoplasms

Objective:To explore the obstacle factors and promoting factors of exercise guidance and implementation after breast cancer surgery, so as to provide reference and basis for the formulation of exercise program after breast cancer patients.Methods:Using the objective sampling method, a total of 10 medical staff and 10 patients with breast cancer after operation from the breast surgery department of the Seven People's Hospital Affiliated to Shanghai University of Traditional Chinese Medicine hospital in Shanghai were selected as research objects from July to September 2022. A semi-structured interview was conducted. The interview outline was developed based on the knowledge transformation framework, and the results were analyzed by the content analysis method.Results:The study extracted three levels of influencing factors. The system level included caregiver workload, educational materials, norms and responsibilities and trust in evidence-based evidence. The practitioner level included the level of exercise knowledge, the concern about patient compliance and the emphasis on exercise. The patient level included sports knowledge level, personality and interest, venous line, adverse reaction of treatment, psychological state, weather environment, attention to health and concern of family members.Conclusions:The medical staff need to consider the postoperative exercise guidance for breast cancer patients from the department, themselves and patients, and should fully consider the patients' conditions and subjective experience to improve the patients' exercise compliance.

Objective:To analyze the efficacy and safety of enzyme replacement therapy (ERT) in Chinese patients with Fabry disease.Methods:A retrospective analysis of the clinical manifestations, genetic variations, family screening, treatments and adverse reactions was conducted in five patients with Fabry disease admitted to the First Affiliated Hospital of Zhejiang University College of Medicine from July 2020 to May 2021. The dosage of agalsidase β was 1 mg/kg by intravenous pump once every 2 weeks.Results:Five male patients with median age of 37 years old (29-51 years old) were diagnosed based on clinical features, family history, α-galactosidase A (α-Gal A) activity, genetic analysis results and kidney biopsy. The clinical manifestations varied in these five patients. All patients had abnormal electrocardiogram, abnormal cardiac ultrasonography and abnormal urinalysis results, three experienced acroparaesthesia during childhood (one patient had persistent pain until adulthood), three had cutaneous angiokeratoma, four had renal insufficiency, four had hypohidrosis, four had diarrheas, four had cornea verticillata and two had high-frequency hearing loss. Two missense mutations of the GLA gene were identified: c.272T>C(p.I91T) and c.868A>G(p.Met290Val). Two nonsense mutations were c.1024C>T(p.Arg342*) and c.838C>T(p.Gln280*). Furthermore, the frameshift mutation c.348del p.(Ile117Phefs*4) was detected, which was not included in the known database, presented with classical Fabry disease. There was no serious adverse reaction during agalsidase β infusion in 5 patients. ERT reduced the plasma globotriaosylsphingosine (lyso-GL-3) levels after treatment of 2-10 months ( P<0.05), and the long-term diarrhea symptom were significantly improved. Conclusions:The clinical manifestations of Fabry disease are varied. Severe adverse events rarely occur in patients treated with short-term ERT. Plasma lyso-GL-3 levels decrease significantly after treatment.

Objective:To explore the clinical application value of group A Streptococcus (GAS) antigen rapid detection method in children suffering from GAS infection.Methods:A total of 44 733 children with suspected GAS infection who were admitted to the Outpatient and Inpatient Departments of Shenzhen Children′s Hospital from January 2015 to December 2019.Throat swab specimens from all children were collected, and BinaxNOW Strep A Test reagent was used for GAS antigen rapid detection.Among them, the throat swabs of 346 children were inoculated with blood culture medium for traditional bacterial culture, and then the GAS antigen rapid detection was tested.The sensitivity and specificity of the two methods were compared, and according to the result of the GAS antigen rapid detection, its age, gender and seasonal trends were analyzed.SPSS 19.0 software was applied for statistical analysis of the data.Results:Among the 346 children tested by both methods, the results of bacterial culture were adopted as the reference method, the sensitivity of the rapid detection method for GAS antigen was 89.41%(152/170 cases), and the specificity was 94.32%(166/176 cases) compared with culture methods.A total of 44 733 cases GAS antigen were tested in children in Shenzhen, of which 10 024 cases were positive, with the positive detection rate of 22.41%.The trend of GAS antigen rapid detection was consistent with the five-year trend, with the high positive rate of 3-8 years, of which 4-6 years of positive rate was the highest.The two seasonal peaks were evident each year, with peaks occurring in April-June, and November and January of next year.The detection rate ratio of male and female was 1.74∶1, and the gender difference was significant ( χ2=27.93, P<0.000 1). GAS antigen rapid detection rate in different clinical departments from high to low in order are as follows: dermatology outpatient (52.34%), emergency clinic (47.74%), internal medicine outpatient (37.36%), infectious disease area (19.71%), five-level disease area (10.27%), internal medicine area (8.63%), surgical areas (7.34%) and neonatal areas (0). Conclusions:GAS antigen rapid detection method and bacterial culture method have high coincidence rate, and high sensitivity and specificity, and can be popularized and applied in the diagnosis of GAS infectious diseases in children.GAS detection rate is higher in outpatient emergency department and dermatology clinics.There are obvious differences from seasonal and population (age and gender) in the positive detection of GAS antigen.No neonates were found.

Objective To investigate aluminum content in commercially available flour products in Minhang District, Shanghai, and to assess the risk of dietary aluminum exposure among permanent residents in Minhang. Methods From 2018 to 2019, a total of 227 samples including fried flour products, steamed flour products, puffed products and baked flour products, were collected in Minhang District to detect the aluminum content. The food frequency table was used to investigate the consumption of flour foods by permanent residents in Minhang District and to assess population dietary aluminum exposure risk. Results The average content of aluminum in 227 kinds of flour products was 31.66 mg/kg, and the exceeding standard rate was 3.96%. The over-standard foods were all fried flour products, and the over-standard rate reached 10.47%. Moreover, the aluminum over standard rate of flour products produced by itinerant vendors and restaurants was significantly higher than that of flour products produced by supermarkets and manufacturers. The average dietary intake of aluminum in Minhang residents was 0.18 mg/kg per week, accounting for 9.03% of the provisional tolerable weekly intake (PTWI). The aluminum exposure of the population with high consumption of foods was 1.4 mg/kg per week, accounting for 69.90% of the PTWI. Among the four types of flour products, fried flour products had the highest contribution rate to aluminum exposure. The daily consumption of fried flour products in males was significantly higher than that in females (P<0.05), and the 18-34 age group was significantly higher than the 60 years old and above group (P<0.05). Conclusion The risk of the average dietary aluminum exposure of flour foods among Minhang residents is relatively low, but people with high consumption of foods would be confronted with increased health risks. It is necessary to strengthen the supervision on the use of aluminum-containing additives in itinerant vendors and restaurants, and actively advocate reducing the frequency and the consumption of fried flour products, thus reduce the exposure to the high risk of dietary aluminum in fried flour products

Objective: To summarize the clinical features of leukoencephalopathy with vanishing white matter disease (VWM) in children. Methods: A retrospective cohort study was performed on 54 genetically diagnosed VWM patients in Peking University First Hospital from January 2007 to March 2019. Paper registration form and electronic medical record system were used to collect the data,and the children were divided into five groups according to the age of disease onset:<1 year, 1-<2 years, 2-<4 years, 4-<8 years and 8-<18 years respectively. The progression of motor function, episodic aggravation, epileptic seizures, survival time, brain magnetic resonance imaging (MRI) and genotype features were analyzed and compared. Non-parametric test, χ² test or Fisher′s exact test were used for comparison among groups; Kaplan-Meier survival curve was adopted to delineate the survival status of the children. Results: Fifty-four VWM patients were included in the study, including 34 males and 20 females.The age of disease onset was 2 years and 8 months (ranged from 6 months to 9 years and 7 months). Onset age was less than 1 year in 5 cases; onset age was 1-<2 years in 12 cases; onset age was 2-<4 years in 25 cases; onset age was 4-<8 years, in 10 cases; onset age was 8-<18 years in 2 cases; 94% (51/54) of patients had complaint of motor regression at the first visit; 87% (47/54) of patients suffered from episodic aggravation. Episodic seizures occurred in 43% (23/54) patients. In survivors with disease durations of 1-3 years, in 38% (9/24) patients the disease was classified as grades Ⅳ-Ⅴ by gross motor function classification system (GMFCS). For the onset age 1-<2 years group, 1 patient was classified as GMFCS Ⅳ among 3 survivors with disease durations of 1-3 years. As for the 2-<4 years group, 6 patients were classified as GMFCS Ⅳ-Ⅴ among 15 patients with disease durations of 1-3 years, whereas 1 patient was classified as GMFCS Ⅳ-Ⅴ among 4 patients with disease durations of 1-3 years in the 4-<8 years group. Lesions, liquefaction and diffusion restriction in brain MRI were compared among different groups, and it was revealed that the earlier the age of disease onset was, the more likely the subcortical white matter (frontal lobe P<0.01,temporal and parieto-occipital lobe both P=0.002), internal capsule (anterior limb P<0.01, posterior limb P=0.00) and brain stem (midbrain P=0.001, pons P<0.01) were to be involved. In addition, internal capsule (anterior limb P=0.002, posterior limb P=0.005) and brain stem (midbrain P=0.001, pons P=0.003) showed more diffuse restricted diffusion. Moreover, the subcortical white matter (frontal and parieto-occipital lobe both P<0.01, temporal lobe P=0.005) showed earlier rarefaction. The 1-year and 2-year survival rates of the overall patients were 81% and 75% respectively, while the 15-year survival rate was 45%. EIF2B5 gene variation was the most common, which accounts for 43% (23/54), followed by EIF2B3 (22%, 12/54). Conclusions The majority of VWM patients complained of motor regression at the first visit, episodic aggravation and epileptic seizures are common in the course. Earlier age at onset is associated with more rapid clinical progression, shorter survival time as well as more extensive lesions, liquefaction and diffusion restriction in brain MRI. The most common variant gene is EIF2B5, followed by EIF2B3.

Objective:To detect the blood flow sepectrum of fetal ductus venosus (DV) with color Doppler ultrasound, and to explore the application of the fetal DV in screening the chromosome abnormalities.Methods:The blood flow spectra of fetal DV in antenatal examination normal group (320cases, with 11-40weeks of gestation) and antenatal examination abnormal group (48cases, within 11-35weeks of gestation) were detected with color Doppler and pulse Doppler ultrasound, and chromosome examination was performed in abnormal group.The fetal DV blood flow patterns (S wave, D wave, a wave) were observed, and karyotype analysis was performed in abnormal group.Results:The normal Doppler spectrum of the DV in fetuses showed the same three-phase waves while the abnormal Doppler spectrum of the DV in fetuses showed a wave reserved or absented.A total of 48cases of chromosome in abnormal group were examined;12cases of chromosome abnormality were found in this study (10cases of DV abnormality and 2cases of DV normality) .A total of 36cases of normal chromosomes were found which (11cases of abnormal DV and 25cases of normal DV) .The abnormal detection rate of DV blood flow spectrum in chromosome abnormal group was higher than that in chromosome normal group (χ2=9.723, P=0.002) .Conclusion:The abnormality of fetal DV blood flow spectrum can increase the detection rate of chromosome abnormalities and ultrasonography could be used as the primary basis for screening the chromosome abnormality in early pregnancy..

To summarize the clinical features of leukoencephalopathy with vanishing white matter disease (VWM) in children. Methods A retrospective cohort study was performed on 54 genetically diagnosed VWM patients in Peking University First Hospital from January 2007 to March 2019. Paper registration form and electronic medical record system were used to collect the data,and the children were divided into five groups according to the age of disease onset:<1 year, 1-<2 years, 2-<4 years, 4-<8 years and 8-<18 years respectively. The progression of motor function, episodic aggravation, epileptic seizures, survival time, brain magnetic resonance imaging (MRI) and genotype features were analyzed and compared. Non?parametric test, χ2 test or Fisher's exact test were used for comparison among groups;Kaplan?Meier survival curve was adopted to delineate the survival status of the children. Results Fifty?four VWM patients were included in the study, including 34 males and 20 females.The age of disease onset was 2 years and 8 months (ranged from 6 months to 9 years and 7 months). Onset age was less than 1 year in 5 cases; onset age was 1-<2 years in 12 cases; onset age was 2-<4 years in 25 cases; onset age was 4-<8 years, in 10 cases; onset age was 8-<18 years in 2 cases; 94% (51/54) of patients had complaint of motor regression at the first visit; 87% (47/54) of patients suffered from episodic aggravation. Episodic seizures occurred in 43% (23/54) patients. In survivors with disease durations of 1-3 years, in 38% (9/24) patients the disease was classified as grades Ⅳ?Ⅴ by gross motor function classification system (GMFCS). For the onset age 1-<2 years group, 1 patient was classified as GMFCSⅣamong 3 survivors with disease durations of 1-3 years. As for the 2-<4 years group, 6 patients were classified as GMFCS Ⅳ-Ⅴamong 15 patients with disease durations of 1-3 years, whereas 1 patient was classified as GMFCS Ⅳ-Ⅴ among 4 patients with disease durations of 1-3 years in the 4-<8 years group. Lesions, liquefaction and diffusion restriction in brain MRI were compared among different groups, and it was revealed that the earlier the age of disease onset was, the more likely the subcortical white matter (frontal lobe P<0.01,temporal and parieto?occipital lobe both P=0.002), internal capsule (anterior limb P<0.01, posterior limb P=0.00) and brain stem (midbrain P=0.001, pons P<0.01) were to be involved. In addition, internal capsule (anterior limb P=0.002, posterior limb P=0.005) and brain stem (midbrain P=0.001, pons P=0.003) showed more diffuse restricted diffusion. Moreover, the subcortical white matter (frontal and parieto?occipital lobe both P<0.01, temporal lobe P=0.005) showed earlier rarefaction. The 1?year and 2?year survival rates of the overall patients were 81% and 75% respectively, while the 15?year survival rate was 45%. EIF2B5 gene variation was the most common, which accounts for 43% (23/54), followed by EIF2B3 (22%, 12/54). Conclusions The majority of VWM patients complained of motor regression at the first visit, episodic aggravation and epileptic seizures are common in the course. Earlier age at onset is associated with more rapid clinical progression, shorter survival time as well as more extensive lesions, liquefaction and diffusion restriction in brain MRI. The most common variant gene is EIF2B5, followed by EIF2B3.

Objective To explore the effect of low-frequency electroacupuncture (EA) on neuropathic pain induced by spinal nerve injury and its underlying mechanism.Methods Thirty-two male Sprague-Dawley rats were randomly divided into a normal group,a sham spared nerve injury (SNI) group,an SNI group and an SNI+EA group,each of 8.The rats in the SNI and SNI+EA groups were given SNI surgery,while those of the sham-SNI group only had the sciatic nerve and its branches exposed without any lesion.EA at 2 Hz was applied over the ipsilateral Zusanli and Kunlun acupoints daily for 14 days after the surgery.The ipsilateral paw withdrawal threshold (PWT) was measured,along with protein kinase A (PKA) levels in the dorsal horn of the spinal cord,calcitonin gene-related peptide (CGRP) and substance P (SP) levels along with transient receptor potential V1 (TRPV1).Results Compared to the normal group,the SNI groups all showed significant decreases in their PWTs on the affected side and significant increases in PKA,TRPV1,CGRP and substance P on the affected side.Compared to SNI group,the average ipsilateral PWT in the SNI+EA group increased significantly after EA treatment,while PKA levels,TRPV1,CGRP levels and SP expression all decreased significantly.Conclusion Electroacupuncture at low frequency can effectively relieve neuropathic pain,perhaps through down-regulation of PKA in the spinal cord and by decreasing pain hypersensitivity related to CGRP and SP.