Objective To investigate the clinical value and sonographic features of locally advanced cervical carcinoma using transvaginal color Doppler ultrasound(TVCDU). Methods A retrospctive analysis was carried on 2-D sonograms. And color Doppler flow imaging(CDFI) in 106 cases, confirmed by pathology and surgery. Results In locally advanced cervical carcinoma, the characteristerics on TVCDU were cervical enlargment, cervical masses accompanying with uteral effusion and extracervical infiltrative masses.The rich cervical blood flow signals demonstrated as called “fireball" sign. In early cervical carcinoma,the cervix showed no apparent cervical enlargment. The blood flow signals were increased.The average RI was 0.41. Conclusions TVCDU which is very valuable in judging the infiltration extent and in differential diagnosis preoperatively can be used as a routine examination method before locally advanced cervical carcinoma treatment.

OBJECTIVE:To establish a method for the residues determination of Pb,Cd,Cu,As and Hg in Codonopsis pilo-sula,and evaluate the quality evaluation of C. pilosula of Pingshun County in Shanxi province. METHODS:Microwave diges-tion-inductively coupled plasma mass spectrometry was adopted with KED scanning model,RF power was 1 550 W,sampling depth was 5.0 mm,plasma gas(argon)flow rate was 16.0 L/min,helium partial pressure was 0.1 mbar,argon gas was 0.6 mbar, the vacuum degree of 5×10-8 mbar,branch turbopump speed was 1 000 hz,sampling cone aperture was 1.0 mm,skimmer aperture was 0.5 mm,the spray chamber temperature was 2.7 ℃,the data collection was repeated 3 times. RESULTS:The linear range was 0-20 ng/ml for Pb(r=0.999 3),0-10 ng/ml for Cd(r=0.998 5),0-250 ng/ml for Cu(r=0.998 8),0-20 ng/ml for As(r=0.999 0) and 0-1.0 ng/ml for Hg(r=0.997 9);RSDs of precision,stability and reproducibility tests were lower than 3.0%;recoveries were 95.80%-100.20%(RSD=1.85%,n=6),94.50%-98.00%(RSD=1.26%,n=6),98.52%-102.43%(RSD=1.60%,n=6), 94.90%-98.70%(RSD=2.29%,n=6)and 96.00%-101.00%(RSD=1.84%,n=6);the limits of detection were 0.021 0,0.003 4, 0.043 7,0.115 6 and 0.005 6 ng/kg,respectively. Pb,Cd,Cu,and As were detetcted,and Hg was not detected,the range of total contents was 7.185 2~12.558 0 mg/kg. CONCLUSIONS:The method is simple with good precision,stability and reproducibility, and can be used for the residues determination of Pb,Cd,Cu,As and Hg in C. pilosula;heavy metal residues in C. pilosula in Shanxi Pingshun county does not exceed limit values of national and industry standards.

Objective To conduct the amplification,cloning,bioinformatics analysis,prokaryotic expression and purification of enterovirus 71 VP1 gene segment and to initially confirm the biological activity of the recombinant expression product.Methods A pair of specific primers was designed according to GenBank EV71 sequence,viral RNA as a template was extracted from the throat swab specimens in the EV71 patients.EV71 VP1 gene was amplified by RT-PCR.After enzyme digestion,the expression vector pET28a was inserted.The prokaryotic expression vector of pET28a-EV71 VP1 was constructed.Then the E.coli DH5a transforma-tion was performed.IPTG was adopted for induction expression.The expression results were analyzed by using SDS-PAGE and Western blot.The bioinformatics analysis of the sequenced results was performed by the software.Expressed protein was purified and the plates were coated,ELISA was used to test the VP1 specific IgG antibody in serum samples of EV71 positive and COX A16-positive patients.Results The BLAST alignment showed that the homology of the objective gene EV71 VP1 was 99% com-pared with other strains(JQ766207.1)in GenBank.EV71 VP1 protein was about 32×103 ,which mainly existed in the form of in-clusion body.The bioinformatics analysis showed that EV71 VP1 protein was a hydrophilic protein,without transmembrane region and N-terminal signal peptide sequence,the tertiary structure existed.The ELISA results showed that the specific IgG OD value in EV71-positive patients was(2.425±0.521),OD value in COX A16 positive patients was(1.205 ±0.314),the normal control OD value was(0.353±0.128).The sensitivity and specificity of EV71 VP1 protein detection were 84% and 88% respectively.Conclu-sion The pET28a-EV71 VP1 expression vector is successfully constructed;the preliminary analysis on the serum of the infected patients by ELISA shows that the obtained objective protein has higher sensitivity and specificity,which is initially confirmed to have biological activity and can be further used for the related study on EV71 diagnosis and vaccine.

PurposeTo explore the value of ultrasound radio frequency technique in the quantitative evaluation of common carotid artery intima-media thickness (IMT) and elasticity change in patients with dyslipidemia, and to discuss the influence of high density lipoprotein cholesterol (HDL-C) upon carotid artery structure and elasticity function. Materials and Methods A hundred and fifty patients with abnormal serum lipid were selected as case group and further divided into three groups: group A--50 patients with hyperlipidemia and high HDL-C, group B--50 patients with hyperlipidemia and normal HDL-C, group C--50 patients with hyperlipidemia and low HDL-C. Meanwhile, 50 healthy people were enrolled in the study as control group (group D). The ultrasound radio frequency-data technique was employed to analyze the influence of hyperlipidemia and HDL-C upon IMT, CC,β, PWV and AIx.Results The IMT,β, PWV and AIx in group B were higher than those in group D, but group B had lower CC (the differences with statistic significance,t=5.141, 4.219, 4.898, 4.142 and-3.758,P<0.01). Compared with group A, group B had higher IMT,β, PWV and AIx and lower CC (the differences with statistic significance,t=3.425, 2.807, 2.913, 2.209 and-2.091,P<0.05 orP<0.01). Compared with group B, group C had higher IMT,β, PWV and AIx and lower CC (the differences with statistic significance,t=4.937, 2.195, 3.337, 2.067 and-3.049, P<0.05 orP<0.01). Group A had higher IMT and PWV than group D (the differences with statistic significance,t=2.236 and 2.407,P<0.05), but CC,β, and AIx showed no difference between group A and group D (t=-1.693, 1.845 and 1.967,P>0.05). In patients with hyperlipidemia, the increase of HDL-C led to the reduction of the common carotid artery IMT and wall stiffness and the increase of elasticity and vice versa.Conclusion The ultrasound radio frequency technique could measure the real-time common carotid artery IMT and elasticity automatically and accurately, thus provides reliable quantitative index for the evaluation of early artery structure and change of function in lipid lowering therapy.

Objective To investigate differences in the expression of Ras 1,Rac1 and Rho1 genes between yeast and hyphal phases of Trichosporon asahii (T.asahii),and to explore their roles in the formation of hyphae.Methods The yeast phase and hyphal phase of T.asahii were cultured and served as yeast phase group and hyphal phase group respectively.Total RNA was extracted from the 2 groups,and real -time fluorescence-based quantitative PCR (RT-PCR) was performed to measure the mRNA expression of Ras1,Rac1 and Rho1.Results The hyphal formation rate was significantly lower in the yeast phase group than in the hyphal phase group (0.40％ ± 0.53％ vs.99.33％ ± 0.57％,t =13.93,P ＜ 0.05).When the mRNA expression of Ras1,Rac1 and Rho1 in the yeast phase group was all set as 1,that in the hyphal phase group was 25.17 ± 10.99,16.81 ± 7.80,42.61 ± 18.50,respectively,with significant differences between the two groups in the three parameters (t =3.81,3.51,3.90,respectively,all P ＜ 0.05).Conclusion Ras1,Rac1 and Rho1 genes may participate in the regulation of hyphal formation in T.asahii.

Objective To perform the amplification ,sequencing and prokaryotic expression of APH (3′′)‐Ⅰ and AAC (2′)‐Ⅰgenes from the clinically isolated gzch810 strain(SM gzch810)of Stenotrophomonas maltophilia to provide the basic materials for the next step functional test .Methods The SM gzch810 genome chromosome was extracted ,the APH (3′′)‐Ⅰ ,AAC (2′)‐Ⅰ whole genes were amplified by PCR and sequenced after being cloned into pMD18‐T vector .The recombination were subcloned into pGEX‐4T‐1 vector and the expression of the recombinant APH (3′′)‐Ⅰ and AAC (2′)‐Ⅰ were analyzed by SDS‐PAGE .Results The 800bp and 550bp DNA fragments of APH(3′′)‐Ⅰ ,AAC(2′)‐Ⅰ gene were amplified from SM gzch810 by PCR and sequenced ;the sequence comparison analysis showed that DNA and amino acid sequence identities of APH (3′′)‐Ⅰand AAC (2′)‐Ⅰ genes with other strains were 91% and 95% respectively .The sequence of APH (3′′)‐Ⅰand AAC(2′)‐Ⅰ of SM gzch810 were submitted to GenBank(accession number :HQ315852 and HQ315853);two major protein bands corresponding to the expected recombinant GST‐TP fusion proteins (56 × 103 and 46 × 103 respectively) were identified by SDS‐PAGE .Conclusion APH(3′′)‐Ⅰand AAC(2′)‐Ⅰgene of SM gzch810 are successfully cloned and expressed ,which lays a good foundation for further detecting corresponding antibi‐otic resistance and functional evaluation of above two kinds of recombinant E .coli .

Objective To investigate the effect of sinomenine on the purinergic receptors A2A and P2X7 in endotoxemia mouse model and RAW264.7 macrophage model stimulated by lipopolysaccharide(LPS). Methods BALB/c mice were randomly divided into blank control group, model group and sinomenine group. Thirty minutes after the rats of sinomenine group were pretreated with intraperitoneal injection of sinomenine (40, 80, 160 mg/kg), the mice were given intraperitoneal injection of 15 mg/kg LPS to induce endotoxemia model. The serum levels of tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of purinergic receptor A2A and P2X7 in the liver and spleen were detected by reverse transcription-polymerase chain reaction(RT-PCR). RAW264.7 macrophages were divided into blank control group, LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine(300μmol/L) for 2 h, and then LPS group and sinomenine group were treated with LPS (100 ng/mL) for another 8 hours. TNF-α in the cell supernatant was measured by ELISA, and the expression levels of A2A and P2X7 in RAW264.7 cells were detected by RT-PCR. Results Stimulation with LPS could induce the increase of the mouse serum levels of TNF-α and IL-6 as well as the expression of A2A and P2X7 in mouse liver and spleen, and sinomenine had a counteraction on the above indexes(P<0.05) . In-vitro stimulation with LPS could induce the increase of the content of TNF-α and the expression of A2A and P2X7 in RAW264.7 cells , and sinomenine decreased TNF-α content and P2X7 expression (P<0.05) , but had an effect on enhancing A2A expression. Conclusion Sinomenine suppresses the expression of purinergic receptor P2X7 in mouse spleen and liver as well as in RAW264.7 macrophages, but its effect on the expression of A2A in various tissues and cells varies, whose related mechanism is needed further study.

Objective:To investigate the feasibility and safety of laparoscopic liver tumor resection in children.Methods:The clinical data of 6 children undergoing laparoscopic liver tumor resection from June 2018 to March 2020 in the Affiliated Tumor Hospital, Zhengzhou University were retrospectively analyzed.Results:There were 4 males and 2 females among the 6 cases, with the average age of (7±2)years.All the children were admitted to the hospital with a liver occupying examination due to physical discomfort.Preoperative diagnosis: 3 cases of hepatoblastoma, 1 case of primary liver cancer, 1 case of hepatic adenoma and 1 case of hepatic nodular hyperplasia.All the children successfully completed laparoscopic liver tumor resection without conversion to open surgery.Among them, 3 children obtained left hemihepatectomy, 1 child underwent hepatic left lobectomy, 1 child was given VI hepatectomy, and 1 child accepted hepatic caudate lobectomy.The operation time was(90±9)min, and the average intraoperative blood loss was (83±26) mL.All children had no blood transfusion during or after operation.There were no bleeding, bile leakage, infection and liver failure after operation.All children had no gastric tube before surgery and the fluid diet was given on the first day after operation, and the postoperative median hospital stay was 4(3-5) days.The pathology were consis-tent with the preoperative diagnosis, and the resections were all radical operation resections.The follow-up period was from 2 to 23 months, and all the children recovered well and no recurrence was observed.Conclusions:Laparoscopic liver tumor resection in children is safe and feasible, which can minimize trauma, reduce intraoperative bleeding and shorten the hospital stay without increasing the incidence of postoperative complications.Laparoscopic liver tumor resection in children has certain advantages.

OBJECTIVETo explore the value of the strain means of the quantitative parameters of organization diffusion in determining the degree of liver fibrosis.

METHODSA total of 130 patients with chronic hepatitis B were examined for quantitative analysis of organization diffusion before liver biopsy to analyze the performance of the strain means in liver fibrosis of different degrees. All the cases were confirmed by pathology.

RESULTSThe strain means differed significantly between different stages of liver fibrosis. Using the best cut-off values of 116.2, 111.4, 101.6 and 97.5 for diagnosis of S≥1, S≥2, S≥3 and S=4, the diagnostic sensitivities were 91.2%, 90.8%, 95.8% and 89.7%, with specificities of 90.4%, 98.6%, 93.8% and 95.6% and accuracies of 90.6%, 94.7%, 95.5% and 91.8%, respectively.

CONCLUSIONThe strain mean can provide specific numerical indicators to determine the degree of liver fibrosis.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors, and its features of insidious onset, rapid development, and high recurrence bring great challenges to the long-term survival of HCC patients. Molecular-targeted drugs and immune checkpoint inhibitors have become the research hotspots in the treatment of HCC, and a large number of clinical trials have found that combined therapy has achieved a good clinical effect. This article mainly introduces the application of combined therapy with molecular-targeted drugs and immune checkpoint inhibitors and other combined therapies in HCC and points out that combined therapy related to systemic therapy or local therapy has become the latest research hotspot in HCC treatment.