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.

Stroke has a high incidence and disability rate, and rehabilitation is an effective means to reduce the disability rate of patients. To systematize rehabilitation assessment, which is the foundation for rehabilitation therapy, we summarize the assessment methods commonly used in research and clinical applications, including the various types of stroke rehabilitation scales and their applicability, and related biomedical detection technologies, including surface electromyography (sEMG), motion analysis systems, transcranial magnetic stimulation (TMS), magnetic resonance imaging (MRI), and combinations of different techniques. We also introduce some assessment techniques that are still in the experimental phase, such as the prospective application of artificial intelligence (AI) with optical correlation tomography (OCT) in stroke rehabilitation. This review provides a useful bibliography for the assessment of not only the severity of stroke injury, but also the therapeutic effects of stroke rehabilitation, and establishes a solid base for the future development of stroke rehabilitation skills.
Artificial Intelligence ; Humans ; Magnetic Resonance Imaging ; Stroke ; Stroke Rehabilitation/methods* ; Transcranial Magnetic Stimulation/methods*

An integrated segmentation method for 3D ultrasound carotid artery was proposed. 3D ultrasound image was sliced into transverse, coronal and sagittal 2D images on the carotid bifurcation point. Then, the three images were processed respectively, and the carotid artery contours and thickness were obtained finally. This paper tries to overcome the disadvantages of current computer aided diagnosis method, such as high computational complexity, easily introduced subjective errors et al. The proposed method could get the carotid artery overall information rapidly, accurately and completely. It could be transplanted into clinical usage for atherosclerosis diagnosis and prevention.
Algorithms ; Angiography ; methods ; Carotid Arteries ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; methods ; Ultrasonography

Objective To evaluate the value of dynamic contrast-enhanced MRI (DCE-MRI)in diagnosing benign and malignant solitary pulmonary nodule (SPN)with Meta-analysis.Methods The databases including PubMed,EBSCO,Cochrane Library,Ovid, CBM,VIP,Wan Fang Database and CNKI were searched to select the literatures about the diagnosis of SPN with DCE-MRI.Inclusion criteria were established,according to the validity criteria for diagnostic research published by the Cochrane Methods Group on Screening and Diagnostic Tests .Methodological quality was assessed by using the quality assessment of diagnostic studies (QUADAS-2)instrument.Meta-analysis was performed by Stata 12.0 and Meta-Disc 1.4.According to the results of heterogeneity of the included articles,proper effect model was selected to calculate the pooled sensitivity,specificity,positive likelihood ration,negative likelihood ration,and diagnostic odds ratio.Summary receiver operating characteristic (SROC)curve was drawed and the area under the curve (AUC)was calculated.Results A total of 17 studies involving 1255 lesions were included.The results of Meta-analysis showed the pooled sensitivity,specificity,positive likelihood ration,negative likelihood ration,and diagnostic odds ratio were 0.95,0.81,4.9,0.06 and 85,respectively.The AUC of SROC was 0.97.Conclusion DCE-MRI has relatively high sensitivity and specificity in the differential diagnosis of benign and malignant SPN.

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 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 analyze the clinical characteristics,the complications and the difference of pregnancy outcome of the patients with systemic lupus erythematosus (SLE) during the different timing of gestation so as to guide the rational pregnancy of the patients with SLE.Methods A systematic review of the clinical data of 96 patients with SLE from January 2000 to January 2018,in Nanjing Hospital Affiliated to Nanjing Medical University,was systematically reviewed,including 60 cases of guiding pregnancy group and 36 cases of non guiding pregnancy group.The general situation,clinical characteristics,pregnancy complications and pregnancy outcome were compared and statistically analyzed.Results There was no significant difference in age and pregnancy between guiding pregnancy group and non guiding pregnancy group (P＞0.05).The rate of lupus activity in guiding group during pregnancy was lower than that in non guiding pregnancy group:25.0％ (15/60) vs.100.0％ (36/36),and the difference was statistically significant (P＜0.01).The main clinical manifestations of SLE during pregnancy were thrombocytopenia,lupus nephritis and rash.The other was infection,arthritis and so on.Among them,the incidences of thrombocytopenia,lupus nephritis and rash in guiding pregnancy group were lower than those in non guiding pregnancy group,and the differences were statistically significant (P＜0.05 or ＜0.01).The incidences of preeclampsia,preterm birth,fetal intrauterine growth restriction (IUGR),low birth weight infants (LBW) and fetal distress in the guiding pregnancy group were lower than those in non guiding pregnancy group,but the difference was not statistically significant (P＞0.05).The pregnancy loss rate was lower and the live birth rate of the guiding pregnancy group was significantly higher than those in non guiding pregnancy group:0 vs.33.3％ (12/36),100.0％ (60/60) vs.66.7％ (24/36),and the differences was statistically significant (P＜0.05).In the guiding pregnancy group,the gestational weeks of termination of pregnancy and the weight of the newborn were greater than those in the non guiding pregnancy group (except for those with pregnancy loss):(269.4 ± 8.0) d vs.(250.8 ± 18.5) d,(3 005.2 ± 305.9) g vs.(2 462.5 ± 507.6) g,and the differences were statistically significant (P＜0.05).The rate of cesarean section in guiding pregnancy group was lower than that in non guiding pregnancy group,and the difference was statistically significant (P=0.019).The mortality rate of pregnant women in guiding pregnancy group was lower than that in non guiding pregnancy group,but there was no statistical difference (P=0.133).Conclusions The main clinical features of pregnancy with SLE during pregnancy are thrombocytopenia,lupus nephritis and rash.Disease activity can increase the incidences of thrombocytopenia,lupus nephritis and rash.In the guiding pregnancy group,the rate of lupus activity during pregnancy is significantly lower than that in the non guiding pregnancy group,and the complications during pregnancy are decreased.Lupus activity is an important factor affecting pregnancy outcome.Choosing the remission stage and control period is the key for successful SLE pregnancy.SLE patients have higher risk of complications than normal people,such as preeclampsia,premature birth,IUGR,LBW,pregnancy loss,and so on.SLE patients should strengthen maternal and child monitoring during pregnancy to minimize perinatal complications and improve perinatal quality.

Objective:To study the effects of Jianpi Qinghua Decoction on hepatocyte apoptosis under non-alcoholic fatty liver disease (NAFLD), and to discuss its mechanism.Methods:Totally 29 C57 mice were randomly selected and fed a 60% high fat diet for 16 weeks, while the remaining 6 mice were given regular feed as the normal group. After successful modeling, 12 mice with larger body weight were divided into TCM group and model group using a random number table method, while continuing to receive high-fat feed. The TCM group was orally administered Jianpi Qinghua Decoction 20.961 g/kg. The normal group and model group were orally administered an equal volume of distilled water once a day, with continuous intervention for 4 weeks. The levels of GOT and GPT in blood were detected by ELISA, the deposition of triglyceride in liver was detected by oil red O, and the apoptosis of liver cells was detected by TUNEL fluorescence staining. Western blot was used to detect the expressions of cleaved Caspase-3, Caspase-3, Bax, Bcl-2, JNK and p-JNK.Results:Compared with the model group, the body weight and GPT in the TCM group significantly decreased ( P<0.05), TG deposition was significantly reduced, apoptosis range of liver cells was significantly reduced, and cleaved Caspase-3/Caspase-3, p-JNK/JNK and the expression of Bax significantly decreased ( P<0.05). The expression of Bcl-2 increased ( P<0.05). Conclusion:Jianpi Qinghua Decoction can inhibit JNK protein phosphorylation and effectively reduce liver cell apoptosis in NAFLD mice, which may delay the progression of NAFLD towards cirrhosis and liver cancer.

Objective:To investigate the changes of various cytokines and coagulation function in B cell acute lymphoblastic leukemia(ALL) patients with different CRS scores during CD19-CAR-T cell immunotherapy.Methods:87 patients with B-ALL hospitalized in the First Affiliated Hospital of Soochow University and 30 normal controls were enrolled into this study from July 2018 to October 2020. The age of the patients was 32(20, 56) years old and 36(41.4%) were female. All these coagulation indicators, prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer, fibrinogen (Fg) were analyzed by automatic blood coagulation in B-ALL patients before and after treated with CAR-T cell. The ratio of CD19-CAR-T cells and the expression of IL-6, IL-10, IFN-γ, TFN-α, IL-2, IL-4, and IL-17A were analyzed using flow cytometry. The patients′ clinical parameters were detected, and the CRS classification of severity was made according to the standard of consensus.Results:Patients with CRS>3 had prolonged PT and APTT, increased D-dimer, and decreased fibrinogen ( P<0.05). The levels of cytokines of IFN-γ, IL-6, and IL-10 were significantly higher in patients with CRS>3 than that in controls ( P<0.05).The D-dimer level is positively correlated with IL-10. Conclusion:Patients with severe CRS grading have significant coagulation dysfunction in CD19-CAR-T cell immunotherapy. Cytokines IFN-γ, IL-6, and IL-10 may affect coagulation function and CRS grading during CD19-CAR-T cell immunotherapy.

Objective:Based on the degradation of skeletal muscle protein MuRF mediated by pFoxO1 when insulin resistance occurs, this paper explores the content change of skeletal muscle protein and the effect of Jianpi Qinghua formula when insulin resistance occurs.Methods:C57 mice were fed with high-fat food and made as the model of obesity accompanied by insulin resistance. Then they were divided into model group, Jianpi Qinghua formula group and metformin group according to random number table method with 10 mice in each group. Jianpi Qinghua formula group was orally administered with water decoction 20.961 g/kg, and the metformin group was orally administered with metformin suspension 18.498 g/kg, once a day for 12 consecutive weeks. Intraperitoneal Glucose Tolerance Tests (IPGTT) was used after the model was established and intervened respectively. The relative protein content of pFoxO1, FoxO1, MuRF, MyoD and myosin were detected by Western blot method, and the localization of MyoD and myosin was detected by immuno-histochemistry.Results:Compared with the model group, the blood glucose of IPGTT at 0 min, 60 min and 120 min of both Jianpi Qinghua formula group and Metformin group decreased ( P<0.05). Compared with model group, the ratio of pFoxO1/FoxO1 protein expression level (0.27±0.07, 0.24±0.14 vs. 0.05±0.03) of both Jianpi Qinghua formula group and Metformin group increased ( P<0.05), and the relative expression of MuRF protein (1.22±0.42, 1.15±0.32 vs. 3.21±0.35) of both Jianpi Qinghua formula group and Metformin group decreased ( P<0.05). The relative protein expression of MyoD (1.42±0.45 vs. 0.40±0.11) and myosin (0.80±0.11 vs. 0.51±0.08) relative protein expression of Jianpi Qinghua formula group was significantly higher than that of model group ( P<0.05). Immunohistochemical staining showed that MyoD (5.06±1.72 vs.2.28±0.83) and myosin (60.28±7.47 vs. 39.77±3.34) of Jianpi Qinghua formula group significantly increased compared with model group ( P<0.05). Conclusion:Jianpi Qinghua formula could effectively increase the content of skeletal muscle protein, enhancing the phosphorylation of FoxO1 in skeletal muscle and the inhibition of MuRF degradation pathway.