Objective To detect the changes of cell proliferation and IFN-γsusceptibility of human pancreatic can-cer cells after suppressor of cytokine signaling-1 (SOCS1) gene silencing, and to explore the SOCS1 as the target of anti-tu-mor therapy through enhancing the function of IFN-γ. Methods Western blot assay, PCR and real-time PCR were used to verify the down regulation of SOCS1 in human pancreatic cancer cell (PANC1) after transfection;subsequently, PANC1 was stimulated with IFN-γ. Western blot assay was also used to detect the expression of signal transducers and activators of tran-scription (STAT)1 and phosphorylation STAT(pSTAT)1;and the change of IFN-γsusceptibility was detected by MTT assay. Real-time PCR was used to detect the mRNA of interferon regulatory factor-1(IRF-1). Flow cytometry was used to detect the cell cycle. Results The expression levels of SOCS1 mRNA and protein were significantly decreased in small hairpin SCOS1 (shSOCS1) transfected PANC1 cells. After the silence of SOCS1, the expression levels of IRF-1 and pSTAT1 in-creased significantly (P<0.05), and the median inhibitory concentration(IC50)of IFN-γfor PANC1 cells decreased signifi-cantly (P<0.01). The cell count of shSOCS1 cells dropped significantly compared with that of control group after the SOCS1 silencing for 72 hours (P<0.05). The cell cycle arrest was promoted at the G0/G1 phase, but the percentage of cells in S phase and G2/M decreased compared to that of control groups (P<0.05). Conclusion After the inhibition of SOCS1 gene expression, the proliferation ability of human pancreatic cancer cell line PANC1 decreased, and the sensitivity of PANC1 cells to IFN-γwas enhanced.

A method based on in situ formed ionic liquids microextraction-ultra performance liquid chromatography(ISFILM-UPLC) was established for the determination of vancomycin in human serum.The samples were pretreated by vortex and centrifugation.[C6MIM] [Br] and NaPF6 were applied as the extracting agent,and Metronidazole as the internal standard.Methanol was applied to redissolve the precipitate.The separation was carried out on a BEH C18 column,and the mobile phase consisted of methanol and 0.05 mol/L monopotassium phosphate solution.The flow rate was 0.2 mL/min,the injection volume was 5 μL,and the column temperature was 18 ℃.The key parameters of this method such as the specificity,accuracy and precision were validated.This method showed good correlation with the immunoassay.It shows great potential for the application in clinical practice.

Objective:To explore the value of HDlive Flow in prenatal diagnosis of the velamentous placenta.Methods:A total of 2 723 pregnant women underwent prenatal ultrasonography in the second trimester and delivered at Zhejiang Provincial People's Hospital from January 2019 to January 2020, and 48 of them were diagnosed as having velamentous placenta confirmed by postpartum clinical and pathological examination and were included in this retrospective analysis. Two-dimensional echocardiography-color Doppler flow imaging (2D-CDFI) and HDlive Flow were both performed during the prenatal ultrasound examination. The sonographic features of velamentous placenta by HDlive Flow were summarized and the prenatal detection rate between two methods were compared using Chi-square test. Results:The incidence of velamentous placenta was 1.8% (48/2 723) in our hospital. Out of the 48 enrolled cases, 45 were diagnosed by HDlive Flow with a detection rate of 93.8% (45/48), three of them were complicated by vasa previa; the other three cases were misdiagnosed as battledore placenta. There were 38 cases diagnosed by 2D-CDFI with a detection rate of 79.2% (38/48), which was lower than HDlive Flow ( χ2=4.360, P=0.037); the other ten cases were misdiagnosed as battledore placenta. The sonographic features by HDlive Flow were as follow: (1) Umbilical cord attached to fetal membranes outside the placenta in 41 cases with the umbilical vessels distributing along the fetal membrane in a mesh pattern; (2) In three cases, the umbilical cord insertion was located on fetal membranes at the edge of placenta; (3) One case was shown that umbilical cord and the branches of umbilical vessels were inserted into the placenta in a "λ" shape. Conclusions:The anatomy of the umbilical cord, umbilical blood vessels and placenta can be directly shown under HDlive Flow, which can improve the prenatal detection rate of the velamentous placenta.

Surface plasmon resonance (SPR) sensor technology,with its features of real-time,fast,no need for labeling,no background interference and non-destructive to samples,etc.,has been widely used in the field of biotechnology,medicine,environmental science and drug detection.This article gives an overview of the recent popular studies and their progress in SPR technology is reviewed,especially giving a detailed overview of the surface modification technique,related hyphenated techniques and application of SPR in clinical examination.Hyphenated techniques is discussed from three aspects of molecular imprinting technique,SPR based immunoassay and nucleic acids SPR biosensor as well.

Sepsis is a life-threatening organ dysfunction caused by dysregulated host responses to infection. Despite decades of research, it remains the leading cause of death in intensive care units (ICUs). None of the current treatment, including antibiotics, organ protection and liquid resuscitation, is specifically effective for sepsis. Immunosuppression is one of the currently accepted pathogenesis and immunotherapy is one of the hot spot of current sepsis research. Immune related treatments include restricting the release of pathogen toxin and its removal, controlling the excessive inflammatory reaction and apoptosis inhibition, etc. Numerous pre-clinical studies using immunomodulatory agents such as interleukin-7 (IL-7), anti-programmed cell death-1 (PD-1) antibody and others, have demonstrated reversal of T cell dysfunction and improved survival resulting from reviewing recent advances in immunotherapy of sepsis. Therefore, immunotherapy may be a new way of sepsis treatment.

Objective To investigate the effects of human papilloma virus 16 E6 (HPV16 E6) on endoplasmic reticulum (ER) stress-autophagic response in the cervical cancer C33A cells.Methods Polymerase chain reaction was used for detecting the integration of HPV DNA.The eukaryotic expression vector of HPV16 E6 was constructed and transfected via lipofectamine into C33A cells.Experimental cells were classified into 3 groups:pcDNA3.1--HPV16 E6 group,pcDNA 3.1-group and C33A group.Western blot was used to measure expression of protein of HPV16 E6,Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 in transfected cells.Results here was no HPV DNA integration in C33A cells that were confirmed as the intervention cells.Eukaryotic expression vector pcDNA3.1--HPV16 E6 was constructed successfully.The eukaryotic expression vector pcDNA3.1--HPV16 E6 significantly improved the expression of protein of HPV 16 E6 in C33A cells.The protein expression of Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 were significantly improved after transfection with vector pcDNA3.1 +-HPV16 E6 (P ＜ 0.05).Furthermore,LC3 Ⅱ protein level was reduced by treatment with ER stress inhibitor.Conclusion HPV16 E6 can improve autophagy through the ER stress pathway,and this response may play an important role in the process of HPV16 E6 inducing cervical cancer,providing one of the new strategies for gene therapy of cervical carcinoma.

Objective:To evaluate our self-designed easily-breaking compression bone bolt plus bone plate in the treatment of tibial plateau fracture.Methods:From July 2018 to December 2018, 12 patients with tibial plateau fracture were treated at Emergency Center of Trauma, The Third Hospital of Hebei Medical University using our self-designed easily-breaking compression bone bolt plus bone plate. They were 8 males and 4 females, aged from 20 to 65 years (average, 45.6 years), with 6 left sides and 6 right sides injured. According to Schatzker classification, there were one case of type I, 3 cases of type Ⅱ, 3 cases of type Ⅲ, 2 cases of type Ⅳ, one case of type Ⅴ and 2 cases of type Ⅵ. The width of tibial plateau was measured and compared before and after operation on X-ray films. Operation time, blood loss and fracture union time were recorded. Loss of reduction and postoperative complications were followed up. Knee function was evaluated at the final follow-up by Rasmussen scoring.Results:For this cohort, operation time averaged 54.6 min, blood loss 25 mL, and fracture union time 17.2 weeks. No nonunion or delayed union was observed. The postoperative width of tibial plateau was (78.9±7.2) mm, significantly narrower than the preoperative value [(87.4±6.1) mm] ( P < 0.05). No loss of reduction or surgical complications occurred postoperation. Deep venous thrombosis of the lower extremity developed in 2 patients but recovered after treatment. The Rasmussen scoring for knee joint function at the final follow-up yielded 9 excellent, 2 good and one poor. Conclusion:In treatment of tibial plateau fractures, our self-designed easily-breaking compression bone bolt plus bone plate can restore width of tibial plateau, compress fracture fragments tightly and allow for early exercise, leading to fine functional recovery of the knee joint.

Objective:To explore the factors related to functional recovery of the knee in patients with tibial plateau fracture complicated with intercondylar ridge fracture.Methods:Included in this retrospective study were 63 patients who had been treated at Emergency Center of Trauma, The Third Hospital of Hebei Medical University from January to June 2019 for tibial plateau fracture complicated with intercondylar ridge fracture. They were 43 males and 20 females, aged from 18 to 66 years (average, 43.9 years). According to our comprehensive classification of tibial plateau fractures, there were 27 simple ones (20 cases of type Ⅰ and 7 cases of type Ⅱ) and 36 complicated ones (16 cases of type Ⅲ, 17 cases of type Ⅳ, one of type Ⅴ and 2 cases of type Ⅵ). To screen out major influencing factors, a multiple linear regression model was used to analyze the 12 factors that might affect functional recovery of the knee by the Hospital for Special Surgery (HSS) scoring: age, gender, occupation, fracture type, way of medical payment, cause of injury, operation method, starting time for rehabilitation, postoperative brace installation, time from injury to operation, length of hospital stay, and presence or absence of a free intercondylar ridge fracture fragment.Results:The 63 patients were followed up for 6.0 to 7.1 months (average, 6.1 months). The HSS knee scores ranged from 45 to 100 points (average, 92.4 points). The HSS knee scores were significantly different between different fracture types ( P<0.05). The HSS scores were significantly higher for the patients without a free intercondylar ridge fracture fragment than for those with ( P<0.05). Multiple linear regression model analysis of the 5 variables with P<0.02 in the single factor analysis (age, fracture type, way of medical payment, hospital stay and presence or absence of a free intercondylar ridge fracture fragment) and HSS scores showed that only fracture type and presence or absence of a free intercondylar ridge fracture fragment had a significant impact on the knee function. Their regression equation was Y=125.591-7.790a-17.277b (Y indicates HSS score, a indicates fracture type and b indicates presence or absence of a free intercondylar ridge fracture fragment). Conclusions:The short-term prognosis for tibial plateau fractures of comprehensive types Ⅰ&Ⅱ (simple ones) may be better than that for tibial plateau fractures of comprehensive types Ⅲ, Ⅳ, Ⅴ&Ⅵ (complicated ones). For patients with a free intercondylar ridge fracture fragment on their lateral knee X-ray film, effective reduction and fixation is indicated but is not for those without.

Objective:To analyze the effects of espO gene knockout on the biological characteristics of enterhemorrhagic Escherichia coli (EHEC). Methods:Two-step methods mediated by the suicide plasmid pCVD442-Δ espO and plasmid pTrc99a were used to construct the espO gene-deleted strain (Δ espO) and the complemented mutant (CΔ espO), respectively. HeLa cells were infected with different EHEC strains to analyze the biological functions and lethal effects of espO gene during infection. Results:PCR, electrophoresis and gene sequencing showed that the Δ espO and CΔ espO mutants were successfully constructed. Compared with the wild-type strain, neither the Δ espO nor CΔ espO mutant showed significant difference in growth rate, indicating that the espO gene had no influence on the growth and replication of EHEC. Furthermore, EspO could activate the tumor necrosis factor receptor (TNF)-induced NF-κB signaling pathway, while the effector protein NleB could inhibit the process. EspO could not inhibit the death of HeLa cells induced by TNF or TNF-related apoptosis-inducing ligand (TRAIL) after EHEC infection. Conclusions:In this study, we successfully constructed the espO gene-deleted and complemented mutants of EHEC and preliminarily analyzed the interaction between espO gene and host cells and the effects of espO gene on cell apoptosis during infection, which provided reference for further research on the in vitro biochemical activity and in vivo pathogenic roles of EspO.

Objective To study the effect of bifidobacterium on intestinal tissue of necrotizing enterocolitis (NEC)in newborn rats and its regulation of Wnt/β-Catenin signal pathway.Methods Seventy -five newborn SD rats were randomly divided into 5 groups,and each group had 1 5 rats.Group A was artificial feeding control group;group B was NEC model group;group C was bifidobacterium treatment group;group D was artificial feeding +bifidobacterium control group;group E was rat breast feeding control group.The localization expression of Toll -like re-ceptor 4(TLR4)of ileocecal ileum tissue was detected by immunohistochemical detection,and also the equivalen-tileum tissues were detected for the contents of glycogen synthase kinase -3β(GSK3β)and β-Catenin expression by Wes-tern blot.Comparing the differences of these indicators between the groups,in addition,the data of TLR4,GSK3βandβ-Catenin were analyzed by Bivariate correlations.Results The levels of TLR4 in ileum tissue of 5 groups were 0.36 ±0.03,0.48 ±0.05,0.34 ±0.03,0.37 ±0.04,0.35 ±0.02.The levels of GSK3βin ileum tissue of 5 groups were 0.98 ±0.23,1 .48 ±0.42,0.99 ±0.20,0.56 ±0.1 7,0.60 ±0.1 5.The levels of β-Catenin in ileum tissue of 5 groups were 1 .48 ±0.22,0.64 ±0.55,1 .27 ±0.36,1 .72 ±0.51 ,1 .82 ±0.44.The levels of TLR4 and GSK3βin ileum tissue of group B were significantly increased compared with group E (P <0.05).The levels of β-Catenin sig-nificantly decreased compared with group E (P <0.05).The levels of TLR4 and GSK3βin ileum tissue of group C were significantly decreased compared with group B (P <0.05).The levels of β-Catenin significantly increased com-pared with group B (P <0.05).Negative correlation was observed between the levels of GSK3βand β-Catenin(r =-0.592,P <0.05),while positive correlation was observed between the levels of TLR4 and GSK3β(r =0.295,P <0.05),and negative correlation was observed between the levels of TLR4 and β-Catenin(r =-0.426,P <0.05). Conclusions Bifidobacterium has certain protective effect on the NEC newborn rat intestines,which can reduce the in-cidence of experimental NEC and the severity of intestinal injury.Its effect may be achieved by regulating the Wnt/β-Catenin signal pathway,which decreases the expression of the level of GSK3βand increases the level of repair fac-tor β-Catenin.