Objective To investigates factors affecting the positive rate of blocking antibody treated by paternal lymphocyte immunotherapy in patients with recurrent spontaneous abortion (RSA).Methods From January 2008 to August 2012,326 RSA cases undergoing treatment in Infertility Center of Qilu Hospital were studied retrospectively.Those patients were divided into 2 groups randomly:260 cases in intradermal injection group were administered via bilateral forearm intradermal injections for immunotherapy once 21 days,then the blocking antibody was determined after 2 (23 cases),3 (73 cases),4 (74 cases),5(90 cases) times respectively,while in subcutaneous injection group,the 66 cases were administered via subcutaneous injection once 21 days,the blocking antibody measured after 3 times; In both cases,the blocking antibody was all determined 2 weeks later.The positive rate of blocking antibodies and the rate of successful pregnancy was recorded,and then followed up after the blocking antibody turning positive.Results (1)Positive rate of blocking antibodies:the positive rate of blocking antibodies were 17％ (4/23),58％ (42/73),72％ (53/74) and 84％ (76/90) in the 2,3,4,and 5 times of intradermal injection group,respectively (P ＜ 0.05).In subcutaneous injection group,the positive rate of blocking antibodies was 38 ％ (25/66),which was significantly lower than that in group intradermal injection receiving 3 times immunotherapy (P ＜0.05).(2) The rate of pregnancy:the 176 patients out of 200 patients were pregnant when antibody was positive after immunotherapy,with 71.6％ (126/176)of patients gained successful pregnancy(the length of pregnancy more than 5 months).Conclusions The route and frequency of administration of immunotherapy could influence the positive rate of blocking antibody.The rate of successful pregnancy will be increased after blocking antibody turning positive.

CRP measurement on automated biochemistry analyzer. The method can be used in clinical diagnosis.

Objective To investigate the feasibility of 80 kV and low tube current on brain CT perfusion(CTP).Methods Sixty patients underwent 80 kV CTP examination,and were randomly divided into five groups according to different tube currents,A group (40 mAs),B group (60 mAs),C group (80 mAs),D group (100 mAs),E group (150 mAs).Twelve patients in each group were selected to analyze the parameters of the caudate nucleus and white matter frontals and temporal,including CBV,CBF and MTT.Subjective image quality,CT value,SD,SNR,CNR and radiation dose were measured and calculated in the 5 groups respectively.Results Differences of CBV and CBF in the caudate nucleus and white matter frontals were existed among A group,B group,and E group (P＜0.05).There were no significant differences of CT value among the 5 groups (P＞0.05).For the SD and SNR,E group was better than A-D groups (P＜0.05).The CNR was higher in E group than that in A group (P＜0.05).The subjective image quality score evaluation in C,D groups were not lower than that in E group(P ＞0.05).The ED of A-D groups were all lower than that of E group (P＜0.05).Conclusion 80 kV/80 mAs brain CTP can obtain accurate perfusion parameter and satisfied image quality,and the radiation dose can be reduced by 46.22％ (2.42 mSv).

Objective:To develop the method based on deep learning to predict the dose distribution of breast-conserving postoperative intensity-modulated radiotherapy(IMRT) for breast cancer, and to evaluate accuracy of the prediction model.Methods:The data of 110 left-sided breast-conserving postoperative IMRT for breast cancer patients were reviewed, among them, 80 cases were randomly selected for training set, 10 cases for validation set and the remaining 20 cases were used as test set.Firstly, the four-channel characteristics of the patients′ computed tomography(CT) images, regions of interest, distances between voxel and planning target volume(PTV), and corresponding dose distributions were taken as input data.The established U-Net was used for training and obtaining prediction model which was utilized to perform dose prediction on the test set, in order to verify the influence of the features of distance between voxel and PTV in dose prediction, and to compare the dose prediction result with the actual manual planned dose.Results:By incorporating the features of distance between voxel and PTV, the model achieved higher accuracy in predicting the dose distribution.The dose scores and dose volume histogram(DVH) scores of the testing set, consisting of 20 patients, were 2.10±0.18 and 2.28±0.08, respectively, and the predicted dose distribution was closer to the manually planned distribution( t=2.52, 2.40, P<0.05). The deviation between the predicted doses of the PTV and the organ at risk (OAR) and the manually planned doses were within 4%, the average dose to the contralateral breast was increased by 13 cGy, all of them within the clinically acceptable range. Except for the statistically significant differences in D2, D98( Di represents the dose received by i%of the PTV volume), Dmean(mean dose) of PTV 60 and V5( Vi was the volume percentage of OAR receiving i Gy dose.), Dmeanof the ipsilateral lung ( t=3.74, 2.91, 2.99, 3.47, 2.29, P < 0.05), there were no statistically significant differences in other parameters. Conclusions:The deep learning-based method can accurately predict the dose distribution of breast-conserving postoperative IMRT for breast cancer, and it has been proven through experiments that by incorporating the features of distance between voxel and PTV can effectively improve the prediction accuracy, which helps physicists to improve the quality and consistency of treatment planning.

Objective To investigate the feasibility of morphology and quantitative parameters for evaluation of lymph node metastatic (LNM) potential in papillary thyroid microcarcinoma (PTMC) with dual?energy CT iodine image. Methods The ninety?five PTMC patients (59 patients with LNM, 36 patients without LNM in the neck) whom underwent dual?energy contrast scanning and confirmed by first postoperative pathology results were enrolled in this retrospective study from July 2014 to December 2016. Dual?energy iodine images were obtained by the dual?energy post?processing software. The iodine image morphology of each patient was analyzed, including size, number, aspect ratio, shape, uncompleted enhanced ring sign, nodule which breaks through the thyroid capsule, microcalcification, and enhanced?degree, and diagnostic efficiency of each imaging feature was calculated. The iodine concentration (IC) and CT value of PTMC solid parts were measured in arterial phase and venous phase images respectively, as well as the NICnod (normalized iodine concentration) and normalized CT value. The independent sample t test was performed to compare quantitative parameters between the LNM group and non?LNM group. ROC curves with quantitative parameters of two phases were used to analyze the diagnostic efficiencies of NIC and normalized CT value. Results Statistically, in the iodine image, the imaging features of LNM group, including multiple lesions, maximum diameter of nodules, aspect ratio, irregular shape, uncompleted enhanced ring sign, and nodule which breaks through the thyroid capsule, had significant differences compared with those of non?LNM group in the neck (χ2 were 14.965,8.724,11.494,8.097,6.324,23.272;P all<0.05), and there were positive correlations with LNM potential (r were 0.397,0.303,0.348,0.309,0.258, 0.495;P all<0.05), the diagnostic efficiency of nodule which breaks through the thyroid capsule was the highest among the imaging features for diagnosis of cervical LNM (sensitivity 77.9%, specificity 72.2%, accuracy 75.8%). During the artery phase, mean NICROI and normalized CT value of non?LNM group were respectively 0.29±0.10,0.33±0.11,and these of LNM group were respectively 0.45±0.15,0.44±0.13.NICROI and normalized CT value of non?LNM group were less than LNM group′s (t=-4.891,－3.462;P all<0.001). During the venous phase, NICROI and normalized CT value of LNM group were higher than non?LNM group′s (t=-3.381,－2.18;P all<0.05). Among all quantitative parameters, the NIC of arterial phase has the highest diagnostic efficacy for the diagnosis of cervical LNM. The area under the ROC curve (AUC) was the largest, 0.814, the sensitivity was 73.5%, the specificity was 79.2%, and the diagnostic threshold was 0.208. Conclusion Dual?energy CT iodine image morphology and quantitative parameters have important clinical value for evaluation of cervical lymph node metastatic potential in PTMC.

AIM:This study examined the inhibitory effect of peiminine on the human colonic adenocarcino-ma cell line SW480 and explored the underlying mechanisms.METHODS:SW480 and human normal colonic epithelial CCD-841CoN cells were treated with different concentrations of peiminine and subjected to the CCK-8 assay to select the optimal treatment time and concentration of the compound.SW480 cell migration and invasion were evaluated by the wound-healing and Transwell assays.Cell cycle progression was analyzed by flow cytometry.The expression levels of cell cycle-related proteins were examined by Western blot.SW480 xenograft tumor model was established in nude mice to ex-amine the effect of peiminine on tumor growth and the expression of cell cycle-related proteins in vivo.RESULTS:Peimi-nine(110 mg/L)significantly inhibited the proliferation of SW480 cells compared with the control group(P<0.01),caused cell cycle arrest at G1 phase,and significantly downregulated the expression of cyclin dependent kinase 2(CDK2),CDK4,CDK6,cyclin D1,p-Rb/Rb,E2F1,E2F3,and E2F4(P<0.05).Peiminine inhibited SW480 xenograft tumor growth,prolonged the survival of model mice,and affected the expression of CDK2,CDK4,CDK6,and cyclin D1 in tu-mor tissues.CONCLUSION:Peiminine promotes G1 phase arrest by down-regulating the expression of CDK2,CDK4,CDK6,and cyclin D1,thereby inhibiting the proliferation of SW480 cells.

AIM:To explore the effect and mechanism of action of the aqueous extract of Fritillaria ussuriensis(FU-AE)against nonalcoholic fatty liver disease(NAFLD).METHODS:The association between Fritillaria ussuriensis Maxir.(FU)and NAFLD was analyzed by network pharmacology.A mouse model of NAFLD was induced in mice by high fat diet(HFD)+10%fructose drinking water,and three doses of Fritillaria ussuriensis aqueous extract were given to the mice for intervention.Colorimetric assay was used for detection of aspartate aminotransferase(AST),alanine aminotrans-ferase(ALT),triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)levels in the serum of experimental mice.Hematoxylin and eosin staining was used to as-sess the pathological and histological changes in the liver of mice and to clarify the anti-NAFLD effect of aqueous extracts of Fritillaria ussuriensis.Liver tissue proteins were extracted,and expression of proteins related to the AMP-activated pro-tein kinase(AMPK)/acetyl-CoA carboxylase(ACC)pathway was detected by Western blot to clarify the mechanism of an-ti-NAFLD action of Fritillaria ussuriensis.The microbial composition of cecum contents was explored using 16S rRNA se-quencing to reveal the modulatory effect of the aqueous extract of Fritillaria ussuriensis on the structure of intestinal flora in mice with nonalcoholic fatty liver disease.RESULTS:Aqueous extract of Fritillaria ussuriensis(high dose)ameliorated exogenous adipocyte infiltration in the liver of mice with NAFLD(P<0.05).AST,ALT,TG,TC and LDL-C levels were significantly decreased(P<0.05)and HDL-C levels were significantly increased(P<0.05)in the high-dose group.Aque-ous extract of Fritillaria ussuriensis(high dose)significantly increased expression of phosphorylated AMPKα,AMPKα,and phosphorylated ACC in the livers of the model mice(P<0.05),significantly reduced expression of ACC(P<0.05),and significantly increased the relative abundance of the potentially beneficial bacteria Faecalibaculum rodentium,Lacto-bacillus johnsonii,Akkermansia muciniphila(P<0.05).CONCLUSION:Aqueous extract of Fritillaria ussuriensis may ameliorate NAFLD in mice by activating the AMPK/ACC pathway and modulating the structure of intestinal flora.

Objective:To investigate the current situation of the use of transjugular intrahepatic portosystemic shunt (TIPS) for portal hypertension, which should aid the development of TIPS in China.Methods:The China Portal Hypertension Alliance (CHESS) initiated this study that comprehensively investigated the basic situation of TIPS for portal hypertension in China through network research. The survey included the following: the number of surgical cases, main indications, the development of Early-TIPS, TIPS for portal vein cavernous transformation, collateral circulation embolization, intraoperative portal pressure gradient measurement, commonly used stent types, conventional anticoagulation and time, postoperative follow-up, obstacles, and the application of domestic instruments.Results:According to the survey, a total of 13 527 TIPS operations were carried out in 545 hospitals participating in the survey in 2021, and 94.1% of the hospital had the habit of routine follow-up after TIPS. Most hospitals believed that the main indications of TIPS were the control of acute bleeding (42.6%) and the prevention of rebleeding (40.7%). 48.1% of the teams carried out early or priority TIPS, 53.0% of the teams carried out TIPS for the cavernous transformation of the portal vein, and 81.0% chose routine embolization of collateral circulation during operation. Most of them used coils and biological glue as embolic materials, and 78.5% of the team routinely performed intraoperative portal pressure gradient measurements. In selecting TIPS stents, 57.1% of the hospitals woulel choose Viator-specific stents, 57.2% woulel choose conventional anticoagulation after TIPS, and the duration of anticoagulation was between 3-6 months (55.4%). The limitation of TIPS surgery was mainly due to cost (72.3%) and insufficient understanding of doctors in related departments (77.4%). Most teams accepted the domestic instruments used in TIPS (92.7%).Conclusions:This survey shows that TIPS treatment is an essential part of treating portal hypertension in China. The total number of TIPS cases is far from that of patients with portal hypertension. In the future, it is still necessary to popularize TIPS technology and further standardize surgical indications, routine operations, and instrument application.

OBJECTIVE To establish the fingerprint and multi-component content determination method of Crataegus pinnatifida leaves from different producing areas， and to evaluate the quality of C. pinnatifida leaves and screen the differential markers. METHODS Seventy-eight batches of C. pinnatifida leaves were collected from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province. High-performance liquid chromatography （HPLC） and Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints and conduct similarity evaluation. Grey correlation analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 19.0， MetaboAnalyst 5.0 and SIMCA 14.1 software. The differential markers affecting the quality of C. pinnatifida leaves were screened with variable importance in the projection （VIP） value greater than 1 and the error line not exceeding the origin as the criterion. Using vitexin rhamnoside as an internal reference， the contents of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin in 78 batches of C. pinnatifida leaves were determined by the same HPLC combined with quantitative analysis of multi- components by single-marker （QAMS）， and the results were compared with external standard method. RESULTS Eight common peaks were calibrated in the fingerprints for 78 batches of C. pinnatifida leaves from 4 producing areas. Five known components were identified， including chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， vitexin rhamnoside （peak 4）， hypericin （peak 7） and isoquercetin （peak 8）； their similarities ranged from 0.871 to 0.998. Average relative correlations of samples from Chengde of Hebei Province， Huludao of Liaoning Province， Yuncheng of Shanxi Province and Linyi of Shandong Province were 0.538， 0.528， 0.462 and 0.435， respectively. CA and PCA showed that the samples from Chengde of Hebei Province and Huludao of Liaoning Province were roughly classified into one category， while the samples from Linyi of Shandong Province and Yuncheng of Shanxi Province were roughly classified into one category； VIP values of peak 1， 2， 3 and 5 were all greater than 1. By QAMS， the relative correction factors of chlorogenic acid， glucosylvitexin， hypericin and isoquercetin were 0.401， 0.993， 1.670 and 1.615 （RSD＜2%）. Compared with external standard method， except for isoquercetin in the two batches of samples （S39 and S41）， there was no significant difference in the content of each component in other batches of samples （the relative deviations≤ 5%）. CONCLUSIONS The established fingerprint and QAMS method are simple to operate and can be used to evaluate the quality of C. pinnatifida leaves. The sample from Chengde of Hebei Province is relatively good in quality. Chlorogenic acid （peak 1）， glucosylvitexin （peak 3）， and the corresponding components of peaks 2 and 5 may be differential markers affecting the quality of C. pinnatifida leaves.

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C