Objective To investigate the effects of respiratory training on motor function in patients with acute stroke. Methods From 2012 to 2014, 80 patients with acute stroke were randomly assigned into treatment group and control group equally. The control group re-ceived routine rehabilitation training, while the treatment group received respiratory training in addition. All the patients were assessed with Fugl-Meyer Assessment (FMA) and modified Bathel Index (MBI) before and eight weeks after treatment. Results The scores of FMA and MBI improved more in the treatment group than in the control group (t>3.938, P<0.001) after treatment. Conclusion Respiratory training may promote the recovery of motor function in acute stroke patients.

Objective To analyze the nucleotide sequences and genetic polymorphisms of UL138 gene of low passage human cytomegalovirus ( HCMV) strains isolated from infants in Guangzhou province. Methods The low passage strains of HCMV were isolated from urine samples of 10 infants with HCMV in-fection in Guangzhou province and identified by multiplex PCR.The UL138 genes were amplified, cloned and identified with sequencing.The sequences were analyzed together with the homologous sequences of 10 clinical isolates published in GenBank.The sequences of UL138 genes were analyzed by using bioinformatics softwares for investigation of the post-translational modification sites, isoelectric points and second structures of UL138 proteins.Results Three low passage strains of HCMV ( D2, D3 and D52) were isolated from in-fants with congenital HCMV infection.The complete sequences of UL138 genes of the three strains were sub-mitted to GenBank after sequencing identification with the GenBank accession numbers of DQ180375, DQ180387 and DQ180359, respectively.The UL138 gene sequences of the three clinical isolates were high-ly conservative.Among the 841 base pairs of the UL138 gene sequences, mutations were identified in 16 sites with base substitution, no any insertion and deletion mutation was found.The 16 mutations resulted in 7 amino acid changes.No additional or deleted sites were found with regard to the post translational modifi-cation sites of UL138 protein in all clinical isolates except the Toledo strain.The isoelectric point of UL138 protein was 6.51 for all clinical isolates.Conclusion The UL138 genes and the deduced amino acid se-quences of HCMV strains isolated from infants in Guangzhou were highly conservative, regardless of the poly-morphism of UL138 gene.This study paved the way for further investigation on HCMV infection and its path-ogenic mechanism.

Objective: To investigate the association between dietary selenium intake and hypertension among Zhejiang residents . Methods: By multistage stratified random sampling method,four urban sites and two rural sites out of Zhejiang Province,four communities or villages out of each site,then 20 households out of each community or village were selected,and all the family members of the selected households were recruited as participants. The questionnaire of Chinese Health and Nutrition Survey was used to collect information about socio-demographic characteristics and dietary selenium intake. The blood pressure,blood lipid and other data were collected via physical examination. A multivariate logistic regression model was used to analyze the relationship between dietary selenium intake and hypertension . Results: A total of 1 222 participants with complete dietary selenium intake data were included for analysis. The number（%）of participants with selenium intake higher than the level of estimated average requirement（EAR）,between the levels of EAR and recommanded nutrient intake （RNI）,between the levels of RNI and upper intake（UI）and higher than the level of UI were 729 （59.66%）,151（12.36%）,341（27.91%）and 1（0.01%）,respectively. There were 283 （30.53%）patients with hypertension out of 927 participants examined. The mean amount of selenium intake in patients with hypertention was（43.06±20.96）μg/d,which was significantly lower than（51.56±30.06） μg/d in non-hypertention participants（P<0.05）. After adjusting for age,body mass index,total cholesterol,triglyceride and diabetes mellitus in the multivariate logistic regression model,dietary selenium intake significantly reduced the risk of hypertension（OR=0.985,95%CI：0.978-0.993） . Conclusion About 60% of residents in Zhejiang Province had lower dietary selenium intake than estimated average requirement. Higher selenium intake was associated with lower risk of hypertension.

Objective: To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression. Methods: UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by ³²P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager. Results: UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA. Conclusions UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Objective:To explore the value of gadolinium-ethoxybenzyl- diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced MRI nomogram model for preoperative prediction of Ki-67 expression in hepatocellular carcinoma (HCC).Methods:Data of 85 patients of HCC confirmed by postoperative pathology, who underwent preoperative Gd-EOB-DTPA enhanced MRI between September 2016 and August 2019 in the First Affiliated Hospital of Soochow University were retrospectively evaluated. According to postoperative immunohistochemistry Ki-67 index, the 85 patients were divided into Ki-67 low expression group(Ki-67 index ≤10%, n=20) and Ki-67 high expression group (Ki-67 index >10%, n=65). Clinical data (hepatitis, cirrhosis, etc.), qualitative imaging parameters (tumor margin, capsule, etc.) were compared by χ 2 test and quantitative parameters [lesion-to-normal parenchyma ratio-arterial phase (LNR-AP), lesion-to-normal parenchyma ratio-portal phase (LNR-PP), lesion-to-normal parenchyma ratio-equilibrium phase (LNR-EP) and lesion-to-normal parenchyma ratio-hepatobiliary phase (LNR-HBP)] were compared by independent sample t test. The above statistically significant parameters were included in multivariate logistic regression to identify the independent predictors of Ki-67 high expression and then the nomogram model for predicting Ki-67 expression of HCC was established. Results:alpha-fetoprotein (AFP) tumor margin, arterial rim enhancement between the Ki-67 low expression group and the Ki-67 high expression group had significant differences (χ 2 were 8.196, 10.538 and 4.717, respectively, P<0.05). LNR-AP, LNR-PP, LNR-EP and LNR-HBP between the two groups had significant differences ( t were 2.929, 2.773, 2.890 and 3.437, respectively, P<0.05).The result of multivariate logistic regression revealed that AFP≥20 μg/L, non-smooth tumor margin and low LNR-HBP were the independent predictors of Ki-67 high expression (odds ratio were 4.090, 3.509 and 0.042, respectively, P<0.05).The Gd-EOB-DTPA enhanced MRI nomogram model for predicting Ki-67 expression of HCC was established successfully. The Area under the receiver operating characteristic curve of the nomogram was 0.837 and the corrected predictive curve fitted the ideal curve, which suggested the model had a good predictive efficiency. Conclusion:Gd-EOB-DTPA enhanced MRI nomogram model has great value in preoperative prediction of Ki-67 expression of HCC, which provided a personalized prediction method for Ki-67 expression in patient with HCC.

Objective: To investigate lunch supply of public primary school canteens in Zhejiang Province, and to provide a basis for the scientific guidance of school lunch. Methods: During May to Jun. and Sept. to Oct. in 2019, lunch food supply was weighed and recorded and the number of diners in 44 public primary school canteens were summarized. Each investigation lasted for one week. Results: Food was mainly based on the taste of the students (40.91%) in the school canteen. About 45.45% of the schools referred to the Nutrition Guidelines of School Meals for students meals when making recipes in the school canteen. The supplies of cereals, vegetables, fruits, livestock and poultry meat, fish and shrimp, eggs, milk, soybean nuts, vegetable oil and salt were 109.05, 118.01, 0, 63.96, 9.25, 11.31, 0, 10.68, 10.47, 2.54 g. The supply of vegetable oil was basically the same as the recommended amount ( P >0.05). The supplies of energy, protein, calcium, iron, zinc, vitamin A, vitamin B 1, vitamin B 2, vitamin C, dietary fiber were 820.84 kcal, 32.79 g, 164.18 mg, 7.84 mg, 4.71 mg, 23.07 μgRAE, 0.41 mg, 0.35 mg, 20.47 mg, 2.34 g, 37.56% of energy from fat and 48.47% of energy from carbohydrate. The supply of vitamin B 1 was basically the same as the recommended amount ( P >0.05). There were no significant differences in all kinds of food and nutrients between urban and rural primary schools ( P >0.05). Conclusion Lunch supply is not optimistic in public primary school canteens in Zhejiang Province, with unreasonable structure and fails to agree with current nutritional recommendations. It is suggested that the scientific guidance of students meals should be carried out according to the survey results combined with the characteristics of local diet.

Objective: To understand current status and related factors of breakfast among primary and secondary school students in Zhejiang Province, so as to provide a scientific basis for improving breakfast habits of primary and secondary school students. Methods: During May to November of 2023, 33 326 students from grade four to six of primary schools and grade one to two of secondary schools were selected from 90 counties and cities in Zhejiang Province by using the stratified cluster random sampling method. General information and breakfast consumption were collected by questionnaire. Multivariate Logistic regression was used to analyze the related factors of breakfast. Results: About 81.29% of the primary and secondary school students reported regular breakfast consumption. The rate of regular breakfast consumption was higher on the school days (92.23%) than on the weekends (85.17%), and higher in primary school students (85.83%) compared to secondary school students (74.71%), with statistically significant differences ( χ 2=827.42, 655.03, P <0.01). About 49.19% of primary and secondary school students had their breakfast within 10 minutes or less, and 83.30% of primary and secondary school students had 3-5 food groups for breakfast. The proportions of students who consumed cereals and potatoes, milk, and eggs were respectively 18.76%, 28.85%, 14.63%. About 22.84%, 28.00 %, 32.60% and 32.23% of the students had no meat, soybeans, vegetables and fruits in their breakfast. The results of multivariate Logistic regression analysis showed that girls, rural area, secondary school, place of living (dormitory, others), migrant parent (one or both outside the hometown), late bedtime (22:00-22:59, 23:00 and later) and late wake up time (9:00 and later) on the weekends were positively correlated with no having breakfast every day ( OR=1.22, 1.40, 1.46, 1.20, 1.20, 1.34, 1.36, 1.41 , 3.51, 2.32, P <0.05). The time of physical activity per day (30-<60, 60-<90, 90-120, >120 min), bedtime (21:00-21:59, 22:00-22:59) and wake up time (6:00-6:59, 7:00-7:59) on school days were negatively correlated with no having breakfast every day ( OR=0.75, 0.64, 0.67, 0.64, 0.77, 0.82, 0.75, 0.67, P <0.05). Conclusions There is a considerable number of primary and secondary school students with irregular breakfast consumption, which are related to multiple factors. Therefore, it is necessary to strengthen nutrition education and improve the behavior of breakfast for primary and secondary school students.

Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.

Objective:To explore the value of different machine learning models based on Gd-EOB-DTPA enhanced MRI hepatobiliary phase radiomics features in preoperative prediction of microvascular invasion (MVI) of hepatocellular carcinoma (HCC).Methods:The data of 132 patients with HCC confirmed by pathology in the First Affiliated Hospital of Soochow University from January 2015 to May 2020 were retrospectively analyzed, including 72 cases of positive MVI and 60 cases of negative MVI. According to the proportion of 7∶3, the cases were randomly divided into training set and validation set. The radiomics features of hepatobiliary phase images for HCC were extracted by PyRadiomics software. The clinical and radiomics features of the training set were screened by the least absolute shrinkage and selection operator (LASSO) regression with 5 fold cross-validation, and then the optimal feature subset was obtained. Six machine learning algorithms, including decision tree, extreme gradient boosting, random forest, support vector machine (SVM), generalized linear model (GLM) and neural network, were used to build the prediction models, and the ROC curves were used to evaluate the prediction ability of the models. DeLong test was used to compare the differences of area under the curve (AUC) for 6 machine learning algorithms.Results:Totally 14 features selected by LASSO regression were obtained to form the optimal feature subset, including 2 clinical features (maximum tumor diameter and alpha-fetoprotein) and 12 radiomics features. The AUCs of decision tree, extreme gradient boosting, random forest, SVM, GLM and neural network based on the optimal feature subset were 0.969, 1.000, 1.000, 0.991, 0.966, 1.000 in the training set and 0.781, 0.890, 0.920, 0.806, 0.684, 0.703 in the validation set, respectively. There were significant differences in the AUCs between extreme gradient boosting and GLM or neural network ( Z=2.857, 3.220, P=0.004, 0.001). The differences in AUCs between random forest and SVM, GLM, or neural network were significant ( Z=2.371, 3.190, 3.967, P=0.018, 0.001,<0.001). The difference in AUCs between SVM and GLM was statistically significant ( Z=2.621 , P=0.009). There were no significant differences in the AUCs among the other machine learning models ( P>0.05). Conclusion:Machine learning models based on Gd-EOB-DTPA enhanced MRI hepatobiliary phase radiomics features can be used to preoperatively predict MVI of HCC, particularly the extreme gradient boosting and random forest models have high prediction efficiency.