OBJECTIVE To establish a ultra-high-performance liquid chromatography-mass spectrum/mass spectrum(UPLC-MS/MS) method for the determination of anlotinib in human plasma and assessment of clinical application. METHODS Zanubrutinib was used as internal standard and the extraction process was performed through protein precipitation method using acetonitrile, followed by separation on an Ultimate XB-C18(100 mm×2.1 mm, 3.0 μm) column using acetonitrile and 10 mmol·L−1 ammonium acetate-0.1% formic acid step-elution gradient. The flow rate was 0.6 mL·min−1 and injection volume was 5 μL. The mass analysis was performed by positive ion electrospray ionization in multiple-reaction monitoring mode, and the mass spectrometer was set at m/z 408.1→339.1 for anlotinib and m/z 472.2→290.1 for internal standard, respectively. The specificity, standard curve and lower limit of quantification, precision and recovery, matrix effect and stability of the method and clinical application were investigated. RESULTS The method was validated over the concentration range of 1.0−100.0 ng·mL−1, with R2=0.998 4. The precision RSD was<9%, the recovery and matrix effect were 104.81%−107.32% and 102.54%−105.26%, respectively, and this method had good stability and was not affected by matrix effect. The method had been used for determined 52 advanced non-small cell lung cancer patients treated with anlotinib. The trough plasma concentration (Ctrough) was measured on day 43 after initiation of anlotinib treatment. Anlotinib Ctrough were higher than lower limit of quantitation (1.0 ng·mL−1) from 52 patients. The plasma concentration of anlotinib Ctrough was (11.38±4.29)ng·mL−1 with 37.66% coefficients of variation, which were shown large inter-patient variability. CONCLUSION This method is high sensitivity, specificity and accurate, and suitable for determination of anlotinib in human plasma.

Objective:To understand the epidemiological characteristics of public health emergency events (PHEE) of varicella in China from 2006 to 2021 and related response performances.Methods:The data of varicella PHEE in 31 provinces of China from 2006 to 2021 were collected through the Public Health Emergency Management Information System, Microsoft Excel 2019 software and SPSS 26.0 statistical software were used to conduct descriptive epidemiological, statistical analysis on the time, area, location distribution, scale and epidemic management.Results:A total of 11 443 PHEE involving 341 048 related cases were reported from 2006 to 2021, with an annual attack rate of 1.78%-3.80% and a total attack rate of 2.33% (341 048/14 624 042). The number of PHEE and related cases of varicella decreased from 1 107 (35 349) in 2007 to 262 (6 884) in 2012 ( Z=-2.40, P<0.001), then increased year by year to 1 318 (42 649) in 2019 ( Z=2.58, P<0.001), with a significant decline since 2020. The varicella PHEE in China presents the seasonal characteristics,the peak is from April to June and from October to December, respectively. The sub-peak of varicella PHEE in eastern China generally appears 1-2 months earlier than in central and western China. Varicella PHEE reports are mainly distributed in eastern China, the attack rate is relatively high in western China, school-reported varicella PHEE was 88.26% of the total reports (10 099/11 443). The epidemic scale of varrcella PHEE typically range from 10 to 29 cases per year among the given outbreaks. The M ( Q1, Q3) of average number of cases, average duration, and average reporting interval of PHEE were 23 (16,35), 20 (14, 26) days, and 9 (5,19) days, respectively, and the reporting interval was positively correlated with the duration ( r=0.854, P<0.001). Conclusions:The varicella PHEE in China from 2006 to 2021 has not been effectively controlled. Schools are the key places to prevent and control varicella PHEE. Improving the sensitivity of varicella PHEE monitoring, strengthening the timely disposal of varicella epidemic, and promoting varicella vaccination are effective measures to prevent and control varicella PHEE.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective:To explore the level of fractional exhaled nitric oxide (FeNO) in preschool children in Beijing and analyze the influencing factors.Methods:Preschool students aged 3-5 in Beijing were selected from May to July 2017.Healthy children were screened through questionnaire survey and on-site physical examination, and their FeNO levels were detected.Results:A total of 317 healthy children were enrolled, including 161 males and 156 females.There was no significant difference in FeNO levels between different genders ( P>0.05). The geometric mean value of FeNO was 8.2 ppb in male(95% CI: 7.0-9.3 ppb)and 8.7 ppb in female (95% CI: 7.4-10.1 ppb). There were statistical differences in FeNO levels among the 3 age groups ( F=4.63, P<0.05). The geometric mean value of FeNO was 7.2 ppb in 3-year-old children (95% CI: 5.5-8.9 ppb), 8.8 ppb in 4-year-old children (95% CI: 7.4-10.2 ppb), and 9.2 ppb in 5-year-old children (95% CI: 7.7-10.7 ppb). The geometric mean value of FeNO of healthy preschool children was 8.5 ppb (95% CI: 7.6-9.3 ppb). There was a positive correlation between FeNO and height ( r=0.135, P<0.05), but there was no significant correlation between FeNO and weight. Conclusions:The geometric mean value of FeNO in healthy preschool children in Beijing is 8.5 ppb (95% CI: 7.6-9.3 ppb)and the 95% reference value is 23.5 ppb.The level of FeNO changes slightly with the increase of age, and height also affects the FeNO.It is highly feasible to use on-line tidal breathing method to measure FeNO of preschool children.

Objective:To construct a Soochow University model of comfortable medical procedures during peri-anesthesia period for patients undergoing radical resection of lung cancer through thoracoscope.Methods:In this prospective study, eight hundreds and sixty patients undergoing radical resection of lung cancer through thoracoscope were enrolled in our hospital in 2018.During peri-anesthesia period, the 860 patients were randomly(by the random number table method)divided into the control group(the group C receiving routine medical procedures, n=430)and the comfort group(the group S receiving the comfortable medical procedures of Soochow University model, n=430). The scores of Visual Analogue Scale(VAS), Richmond Agitation-Sedation Scale(RASS), and Thirsty Analogue Scale(TAS)were recorded at 5 min after extubation(T1), at out of the post-anesthesia care unit(T2), at 1 h(T3), 6 h(T4), 12 h(T5), 24 h(T6), 2 d(T7)and 3 d after surgery(T8)in two groups.The incidences of postoperative sore throat(POST), nausea and vomiting(PONV), catheter-related bladder discomfort(CRBD), thirst and hypothermia were recorded at T2, T6 and T8, respectively.Meanwhile, the peri-anesthesia comfort questionnaire(PCQ)and peri-anesthesia satisfaction questionnaire(PSO)were completed at T6 and T8.Results:The incidences of POST(48.6% vs.16.2%), PONV(24.9% vs.13.0%), CRBD(78.8% vs.20.9%)and thirst(74.9% vs.20.0%)were higher in the group C than in the group S at T2( P<0.05). The comfort score and satisfaction score were lower in the group C than in the group S at T6(3.14±1.04 vs.4.92±1.42, 2.67±0.89 vs.3.30±1.01)and at T8(3.84±1.83 vs.5.05±1.77, 2.74±0.84 vs.3.26±1.06)( P<0.05). Conclusions:The Soochow University model of comfortable medical procedures during peri-anesthesia period for patients undergoing radical resection of lung cancer through thoracoscope has been successfully constructed, and this model can reduce the incidence of complications, and improve the comfort and satisfaction during peri-anesthesia period.

Objective To investigate the therapeutic effect of Jianpi Yiqi, Bushen Fenjing combined with oxaliplatin+CF/5-FU on colorectal cancer.MethodsEighty patients with colorectal cancer who were treated in our hospital from January 2014 to December 2015 were randomly divided into observation group(40cases) and control group (40cases).The control group was given oxaliplatin+CF/5-FU treatment, the observation group in the control group based on the application of Spleen Qi, Bushen fill essence treatment, observed before and after treatment of two groups of serum tumor markers, clinical symptoms and living conditions, changes, record adverse reactions.ResultsThe observation effect of 82.50% was significantly higher than that of the control group (62.50%).The QOL score (37.8±5.6) was significantly higher than that of the control group (33.5±3.1) and the adverse reaction rate was 30.00% (P<0.05).The difference between the two groups was significant (P<0.05).ConclusionThe effect of invigorating spleen qi and tonifying siren solution combined with oxaliplatin+CF/5-FU on colorectal cancer can improve the clinical symptoms and improve the quality of life of patients.

Objective To explore the influence of group B Streptococcus screening during pregnancy and the incidence of the ear‐ly‐onset GBS disease for newborns .Methods Totally 47 cases of pregnant women with premature rupture of membranes (PROM ) , which were GBS positive and accepted antibiotic treatment ,who were chosen as the experimental group .While 73 cases of pregnant women with premature rupture of membranes (PROM) ,which were not accept GBS screening and antibiotic treatment ,were chosen as control group .The neonatal clinical manifestations were observed .The swab specimens were collected from throat and detected of GBS by using PCR method .Results The experimental group showed no occurrence of neonatal group B streptococcal infection , dyspnea ,cyanosis and fever .Totally 7 cases of the control group had group B Streptococcus infection .Totally 2 cases had dyspnea and 2 cases had cyanosis .Totally 4 cases had fever .The neonatal research indicators of these two groups were statistically signifi‐cant differences (P<0 .05) .Conclusion The group B Streptococcus screening during pregnancy would effectively reduce the inci‐dence of neonatal infection of group B Streptococcus .

Objective To observe any effect when eye acupuncture is combined with computer-assisted cognition training to ameliorate cognitive dysfunction after stroke.Methods Stroke patients with cognitive impairment were randomly divided into an eye acupuncture group (n=30),a computer group (n=30) and a combination group (n=30).The eye acupuncture group was given eye acupuncture,the computer group was given rehabilitative cognition training with specialized equipment and the combination group was given both eye acupuncture and the computeraided training.The treatment lasted 2 months.The Loewenstein occupational therapy cognitive asessment (LOTCA)and the modified Barthel index were used to evaluate the patients' cognitive function and daily life ability before and after treatment.Results Before training there was no significant difference in average LOTCA and MBI results among the three groups.After 2 months of training,all three groups had significantly higher scores in both evaluations.Moreover,the combination group's average score on organization ability,perceptual ability,thinking operation ability and concentration,and also their average total score and MBI score were significantly better than those of the other two groups.Conclusions Eye acupuncture combines synergistically with cognitive rehabilitation training to ameliorate cognitive dysfunction after stroke.Together they promote cognition,ADL ability and an early return to normal family and social life better than either alone.

Objective To characterize the mutation of phenylalanine hydroxylase (PAH) gene in patients with phenylketonuria(PKU) in Ningxia area,China.Methods Seventy-three children diagnosed with PKU at the Child and Maternal Healthcare Hospital of Ningxia Hui Autonomous Region between January 2010 and June 2013,and 100 non-PKU children randomly chosen from children with normal results in PKU screening were enrolled in the study.Venous blood was collected and the PAH gene sequence was determined by direct DNA sequencing after amplification with the polymerase chain reaction technique.The new gene mutations were defined based on the national and international literature search and databases.The source of the newly discovered mutations was also measured by examining and sequencing the blood samples of their parents.The Chi-square test was used for statistical analysis.Results Among 146 alleles of the 73 PKU children,the detection rate of mutation of PAH gene was 79.5％ (116/146),including 37 types of mutations occurring in 11 exons other than exon 2 and exon 13.The 37 different mutations included 22 missense mutations (59.5％,22/37),six nonsense mutations(16.2％,6/37),six splice site mutations(16.2％,6/37) and three deletion mutations(8.1％,3/37).p.R243Q(17.1％,25/146),EX6-96A ＞ G (6.8％,10/146),p.R241C(6.2％,9/146),p.R413P (5.5％,8/146),p.Rl11X(4.8％,7/146) and IVS4-1G ＞ A(4.8％,7/146) were found to have a higher mutation frequency.Meanwhile,p.R243Q was the most common mutation among Han and Hui ethnic groups with a frequency of 18.8％(12/64) and 15.9％ (13/82),respectively.In contrast,p.R241C showed a significant higher frequency in the Hui group [9.8％(8/82) vs 1.6％(1/64),x2=4.17,P=0.04].Four new mutations of PAH genes,including p.Q304K,p.H107R,p.F392I and p.N223I,were discovered after literature search and comparative studies.Conclusions PAH gene mutations in children with PKU in Ningxia area are unique and are characterized by the diversity and complexity of mutation occurrence in this ethnic region.

Background and objectiveWIF-1 is an important tumor-suppressing gene in lung cancer, and its encoding protein WIF-1 can reduce proliferation and promote apoptosis by inhibiting Wnt/β-catenin signaling in lung can-cer. hTis study constructs a eukaryotic expression plasmid carrying WIF-1 using FDA-approved clinical plasmid pVAX and explores the anti-tumor effect of pVAX-WIF-1 on A549 lung cancer cellsin vitro andvivo.MethodshTe DNA fragment of human WIF-1 coding sequence was ampliifed by PCR and was cloned into the multiple cloning sites of eukaryotic expression vector pVAX to construct pVAX-WIF-1. A recombinant plasmid was transfected into lung cancer A549 cells, and the expression ofWIF-1 genes was veriifed by Western blot atfer transfection. Subsequently, the effect of pVAX-WIF-1 on cell apoptosis and proliferation was identiifed by MTT assay, staining A549 cells with Hoechst 3235, and lfow cytometry. Finally, the A549 sub-cutaneous xenogratf was established to detect the effect of pVAX-WIF-1 on lung tumor growthin vivo.Results hTe results of restriction enzyme digestion, PCR, and sequencing indicated that eukaryotic expression plasmid pVAX-WIF-1 was successfully constructed. hTe protein expression level of WIF-1 was increased in the transfected A549 cells. Further results showed that transfection with pVAX-WIF-1 signiifcantly inhibited proliferation and promoted apoptosis in A549 cells. Moreover, pVAX-WIF-1 signiifcantly inhibited the tumor growth of the A549 subcutaneous xenogratfin vivo.ConclusionhTe recombinant eukaryotic expression vector pVAX-WIF-1 was successfully constructed. Transfection with pVAX-WIF-1 could signiifcantly inhibit proliferation and promote apoptosis of lung cancer A549 cells and also effectively inhibit the tumor growth of the A549 subcutaneous xenogratfin vivo. Our research can contribute to clinical applications of WIF-1 in lung cancer gene therapy.