Objective To detect and analyze the drug resistance characteristics,phylogenetic typing,and virulence gene distribution of Escherichia coli(E.coli)in blood culture.Methods The strains of E.coli isolated from consecutive non-repetitive blood cultures in our hospital from January 1,2019 to December 13,2020 were collected.The sensitivity of E.coli to 17 antibiotics was determined u-sing the micro-broth method.The bacterial genomic DNA was extracted using the boiling method,and then the arpA,chuA,yjaA,TspE4C2,ArpAgpE and trpAgpC genes were detected by PCR to determine the bacterial phylogroup.The virulence genes,including iutA,fimH,fyuA,kpsMT Ⅱ,cnf1,cvac,hlyA,traT,kpsMT Ⅲ,and PAI,were detected using the multiplex PCR.The differences in drug resistance and virulence gene distribution among different phylogroups were analyzed by the Chi-square test.Results 270 strains of E.coli in blood culture showed high resistance rates to ceftriaxone,compound sulfamethoxazole,ampicillin,ampicillin sulbactam,cefazolin,and ciprofloxacin,all exceeding 50.0％.They had good susceptibility to imipenem,ertapenem,amikacin,and piperacillin tazobactam,with resistance rates all below 5.0％.The most common phylogroups were types B2 and D,accounting for 38.0％and 16.2％,respectively,while type E and hidden branch type I were relatively rare,accounting for less than 1.0％.The virulence gene analysis revealed that the distribution rates of fimH and fyuA genes were the highest,both above 99.0％.The distribution rates of kpsMT Ⅲ,hlyA,and cvaC genes were relatively low,all below 20.0％.The Chi-square test showed that the distribution rates of viru-lence genes such as iutA,fimH,fyuA,kpsMT Ⅱ,cnf1,and PAI in the B2 group were significantly higher than those in the non-B2 group(P<0.05).The distribution rates of iutA,fyuA,kpsMT Ⅱ,cnf1,and PAI genes in the B2 group were significantly higher than those in the D group(P<0.05).Conclusion When treating bloodstream infections caused by E.coli,caution should be exercised in the use of drugs such as ceftriaxone,compound sulfamethoxazole,ampicillin,ampicillin sulbactam,cefazolin,and ciprofloxacin.When bloodstream infections are caused by phylogroup B2 E.coli,middle-stream urine culture should be performed simultaneously to confirm the source of infection and monitor the success rate of treatment.

Enzyme-driven micro/nanomotors consuming in situ chemical fuels have attracted lots of attention for biomedical applications. However, motor systems composed by organism-derived organics that maximize the therapeutic efficacy of enzymatic products remain challenging. Herein, swimming proteomotors based on biocompatible urease and human serum albumin are constructed for enhanced antitumor therapy via active motion and ammonia amplification. By decomposing urea into carbon dioxide and ammonia, the designed proteomotors are endowed with self-propulsive capability, which leads to improved internalization and enhanced penetration in vitro. As a glutamine synthetase inhibitor, the loaded l-methionine sulfoximine further prevents the conversion of toxic ammonia into non-toxic glutamine in both tumor and stromal cells, resulting in local ammonia amplification. After intravesical instillation, the proteomotors achieve longer bladder retention and thus significantly inhibit the growth of orthotopic bladder tumor in vivo without adverse effects. We envision that the as-developed swimming proteomotors with amplification of the product toxicity may be a potential platform for active cancer treatment.

Objective:To evaluate the clinical efficacy of microneedle radiofrequency in the treatment of moderate acne vulgaris.Methods:From August 2018 to August 2020, 393 patients (192 males and 201 females, aged 15-38 years) with moderate acne were enrolled in the Department of Dermatology, the First Affiliated Hospital of Shantou University Medical College and Shantou Chaonan Minsheng Hospital, including 201 patients in experimental group and 192 patients in control group. In the experimental group, microneedle radiofrequency therapy was used once every 2 weeks for 3 times in total. The control group adopted the fire needle, once every 2 weeks, a total of 3 times. The efficacy of both groups was evaluated at week 8.Results:A total of 378 patients were actually completed: 196 patients in the experimental group, and 182 patients in the control group. At the eighth week of follow-up, the total effective rate was 81.12% in the experimental group and 70.43% in the control group. The efficacy of the two groups was statistically different (χ 2=4.42, P<0.05). Conclusions:The efficacy of microneedle radiofrequency therapy in the treatment of moderate acne vulgaris is better than that of fire needle, with good tolerance, short recovery period, few adverse reactions and high compliance, which has clinical promotion value.

Objective : To explore the technology and efficacy of fabrication of a guide and removal of a fiber post by tetrahedron positioning technology at the chair side. Methods: For one patient with acute chronic periapical periodontitis of the left maxillary lateral incisor who needed to have the fiber post removed, the chair side tetrahedral positioning technique was used to make a guide plate to remove the fiber post. Cone beam CT (CBCT) data were imported into the software to design the guide plate for fiber post removal. The guide plate design on CBCT was transferred to a solid model by using tetrahedral positioning technology. The guide plate was made to guide the removal of the fiber post, and then left maxillary lateral incisor root canal was performed. We evaluated the effect of fiber post removal with tetrahedral positioning technology by reviewing the literature. Results : The guide plate made by tetrahedral positioning technology can accurately locate the position and direction of fiber posts at a low cost and with high speed. After the fiber post was removed, the root canal could be dredged by using root canal preparation instruments. After root canal preparation, the root canal was filled with warm gutta-percha to complete the root canal treatment. After 3 months, the apical radiograph showed that the transmission shadow of the apical area was reduced. The results of the literature review showed that the fiber post removal with guide plates provides a predictable result and a lower risk of iatrogenic damage. Minimally invasive treatment can be carried out, and chair time can be reduced. Conclusion On the basis of CBCT data, using tetrahedral positioning technology to make fiber post removal guides can help reduce the risk of fiber post removal and has the characteristics of speed, low cost and short chair side processing. However, the accuracy comparison between tetrahedral positioning technology and 3D printing guides needs further study.

Excessive theta (θ) frequency oscillation and synchronization in the basal ganglia (BG) has been reported in elderly parkinsonian patients and animal models of levodopa (L-dopa)-induced dyskinesia (LID), particularly the θ oscillation recorded during periods when L-dopa is withdrawn (the off L-dopa state). To gain insight into processes underlying this activity, we explored the relationship between primary motor cortex (M1) oscillatory activity and BG output in LID. We recorded local field potentials in the substantia nigra pars reticulata (SNr) and M1 of awake, inattentive resting rats before and after L-dopa priming in Sham control, Parkinson disease model, and LID model groups. We found that chronic L-dopa increased θ synchronization and information flow between the SNr and M1 in off L-dopa state LID rats, with a SNr-to-M1 flow directionality. Compared with the on state, θ oscillational activity (θ synchronization and information flow) during the off state were more closely associated with abnormal involuntary movements. Our findings indicate that θ oscillation in M1 may be consequent to abnormal synchronous discharges in the BG and support the notion that M1 θ oscillation may participate in the induction of dyskinesia.

Objective: To determine the relationship between dengue virus load and clinical characteristics, so as to provide basis for dengue fever prevention and treatment. Methods : The dengue viral load and typing of 120 patients in Gongshu District of Hangzhou from June to November 2017 were detected by real-time fluorescent quantitative RT-PCR;the clinical indicators of these dengue patients were collected and their correlation with the viral load was analyzed. Results: The DNA detection of dengue virus in 120 patients showed that they were all typeⅡ. The median dengue virus load was 3.91×104 copies/mL. All the patients had fever, the average peak temperature was（38.96 ± 0.69）℃. There were 102（85.00%）cases with asthenia;116（96.67%）cases with white blood cell count（WBC）less than 4× 109/L;119（99.17%）cases with platelet count（PLT）less than 100×109/L;114（95.00%）cases with glutamic oxaloacetate transaminase（GOT）more than 40 U/L;81（67.50%）cases with glutamic pyruvate transaminase（GPT）more than 52 U/L;58（48.33%）cases with creatine kinase（CK）more than 210 U/L. There was no significant correlation of dengue virus load with length of hospitalization, peak temperature,duration of fever, WBC,PLT, GOT, GPT and CK（P>0.05）. There were 75（62.50%）severe patients, and their median viral load was 9.29×104copies/mL, which was higher than 5.33×103copies/mL in non-severe patients（P<0.05）. Conclusion The dengue virus load is not related with length of hospitalization,peak temperature,WBC,PLT,GOT,GPT and CK,but with the severity of the disease.

Objective To investigate the mechanism of improvement of gait behavior in PD rat models by low frequency electrical stimulation of pedunculopontine tegmental nucleus (PPTN) by optogenetics method. Methods (1) Twenty-four healthy adult male SD rats were randomly divided into a sham-operated group 1, a lesion group 1 and a photoactivation group (n=8); normal saline was injected into the right medial frontal tract (MFB) of the sham-operated group 1; 6-hydroxydopamine (6-OHDA) was injected into the lesion group 1 and photoactivation group to induce PD models; two weeks after modeling, injection of adeno-associated virus hsynapsin-ChR2-mcherry into the right PPTN of the three groups was performed, and the photoactivation group received blue-ray stimulation by implanting optical fibers into the PPTN at the same time. (2) Twenty-four healthy adult male SD rats were randomly divided into a sham-operated group 2, a lesion group 2 and a photoinhibition group (n=8);normal saline was injected into right MFB of the sham-operated group 2; 6-OHDA was injected into the lesion group 2 and photoinhibition group to induce PD models; two weeks after modeling, injection of adeno-associated virus hsynapsin-NpHR-mcherry into the right PPTN of the three groups was performed, and the photoinhibition group received yellow-ray stimulation by implanting optical fibers into the PPTN at the same time. (3) Three weeks after injection of adeno-associated virus, Catwalk gait analysis was used to assess the behavioral ability of rats in each group. Results (1) As compared with the sham-operated group 1, lesion group 1 had significantly increased front claw spacing and back front claw spacing, and significantly decreased stride length and pressure of damaged lateral and contralateral limbs, and significantly decreased swing speed of contralateral limb (P<0.05); as compared with those in the lesion group 1, the front claw spacing and back claw spacing were significantly shortened, and stride length and pressure of damaged lateral and contralateral limbs were statistically increased in the photoactivation group (P<0.05). (2) As compared with the sham-operated group 2, lesion group 2 had significantly increased front claw spacing and back front claw spacing, significantly decreased stride length of damaged lateral limb, and significantly decreased pressure and swing speed of damaged lateral and contralateral limbs (P<0.05); no significant differences were noted on the front claw spacing and back front claw spacing, pressure and swing speed of damaged lateral and contralateral limbs between lesion group 2 and photoinhibition group (P>0.05). Conclusion The mechanism of low frequency electrical stimulation of PPTN improving gait behavior of PD rat models is related to activation of PPTN neurons.

Objective To investigate the expression of zeste white 10 interactor (Zwint) in primary hepatocellular carcinoma (HCC) and its effect on the prognosis of liver transplantation for HCC. Methods HCC tissues, paracancerous tissues and clinical data of 50 liver transplant recipients for HCC were collected. The expression levels of Zwint messenger RNA (mRNA) and Zwint protein in 20 pairs of HCC tissues and paracancerous tissues of 20 liver transplant recipients for HCC were compared using real-time fluorescence quantitative polymerase chain reaction (PCR), Western Blot and immunohistochemistry (IHC). Two HCC cell lines HepG-2 which interfered with the expression of Zwint successfully were selected as si-Zwint-1 group and si-Zwint-2 group, and the blank control was taken as si-NC group. The cell proliferation and cell cycle of various groups were compared using cell counting kit (CCK) -8 experiment, flat-cloning assay and cell cycle experiment. The consistency of the expression of Zwint and cyclin D1 in HCC tissues and cells was analyzed using Western Blot and IHC. The enrolled patients were divided into high expression group (22 cases) and low expression group (28 cases) based on the median of Zwint protein expression level, and the relationship of the expression level of Zwint protein and clinical characteristics, overall survival rate and disease free survival rate of liver transplant recipients for HCC was analyzed. Results The results of real-time fluorescence quantitative PCR showed that the expression level of Zwint mRNA in HCC tissues was higher than that of paracancerous tissues (P=0.03). The results of Western Blot and IHC showed that the expression level of Zwint protein in HCC tissues was higher than that of paracancerous tissues(both P<0.05).After the Zwint gene of HCC cell line HepG-2 was interfered,CCK-8 and flat-cloning assay showed that the cell proliferation potential was significantly weakened (all P<0.01), and the cell cycle arrested at stage G1(all P<0.05). The expression level of Zwint protein was closely related to tumor diameter and tumor, node, metastasis (TNM) staging (all P<0.05). The overall survival rate of liver transplant recipients for HCC in the high Zwint expression group was lower than that of the low expression group (P=0.02). Conclusions Zwint is highly expressed in HCC tissues, and it can promote the proliferation of HCC cells through regulating cell cycle. The expression level of Zwint is negatively correlated with the prognosis of liver transplantation for HCC.

Objective To explore the mechanism of microRNA (miRNA, miR)-155 in the rejection after liver transplantation in rats. Methods The rats were divided into two groups. In the xenograft model group (rejection group, n=10),the donors were male Lewis rats and the recipients were male BN rats.In the allograft model group(control group, n=10),both the donors and recipients were male Lewis rats.The rat models with orthotopic liver transplantation were established by two-cuff technique in two groups. At postoperative 7 d, the animals were sacrificed for the collection of blood and liver tissue samples. The serum levels of alanine aminotransferase (ALT), total bilirubin (TB), and cytokines of interleukin (IL)-2, IL-4, interferon (IFN)-γ were quantitatively measured. The pathological changes of liver tissues were observed under light microscope. In each group, three liver tissue samples were prepared and subject to high-throughput sequencing. The miRNAs related to rejection were identified for bioinformatics analysis to predict and analyze relevant signaling pathways and genes. Results In the rejection group, the serum levels of ALT and TB were significantly higher than those in the control group (both P<0.01). Compared with the control group, the levels of IL-2 and IFN-γ were considerably up-regulated (both P<0.01), whereas the level of IL-4 was dramatically down-regulated (P<0.01). Pathological examination demonstrated that more evident rejections were observed in the rejection group than the control group. High-throughput sequencing revealed that the expression level of miR-155 was significantly up-regulated in the rejection group, which was 5.89 times of that in the control group. Bioinformatics analysis demonstrated that up-regulation of miR-155 was associated with the mammalian target of rapamycin (mTOR), mitogen-activated protein kinase (MAPK) and T cell receptor signaling pathways. The genes which were probably responsible for regulation included the yeast autophagy related gene 1(ATG1) and its homologous gene ULK2, insulin-like growth factor-1 (Igf-1) and G protein-coupled receptor regulatory gene(Arrb1),etc.Conclusions miR-155 might promote the incidence and progression of rejection after liver transplantation in rats. The involved signaling pathways probably include the mTOR, MAPK signaling pathway and T cell receptor signaling pathway.ATG1,ULK2,Igf-1,and Arrb1 genes may participate in this process.

Objective To establish male rat models for fertility following liver transplantation. Methods Male Sprague-Dawley (SD) rats were used as the donors and recipients of liver transplantation. The donor liver was transplanted with two-cuff technique. Liver transplantation was performed in 15 male SD rats. At 3 weeks after liver transplantation, 5 rats were randomly sacrificed for detection of sperm deformity rate. The remaining male rats were mixed bred and mated with healthy female SD rats at a ratio of 1︰2. General conditions of the rats undergoing liver transplantation were recorded. Liver function parameters were detected after liver transplantation. Postoperative sperm deformity rate was observed. The pregnant status of female rats and health situation of their offsprings was monitored. Results All 15 rats (100%) underwent liver transplantation successfully. Nine rats (9/10) survived longer than 8 weeks. Liver function parameters were normal in male rats following liver transplantation. The sperm deformity rate was ranged from 0.5% to 1.3%. Ten male rats undergoing liver transplantation were mixed bred with female rats at a ratio of 1︰2 for 1 week. All female rats were successfully mated and delivered their offsprings after 3 weeks. The offsprings had no evident physiological deformity. Conclusions Male rat models for fertility are successfully established after liver transplantation, which serve as an animal model to evaluate the fertility performance in male patients undergoing liver transplantation.