Objective To prepare the recombinant Echinococcus granulosus Polo-like kinase 2 (rEgPLK2) protein and evaluate its immunoprotective efficacy against cystic echinococcosis, so as to provide insights into research and development of novel vaccines against echinococcosis. Methods The Polo-like kinase (PLK) protein sequences were retrieved from 12 species in the NCBI protein database, including E. granulosus and E. multilocularis. Multiple sequence alignment was performed using the Clustal Omega program, and structural visualization and homology analysis were conducted using the ESPript 3.2 program. The recombinant plasmid pET-30a-EgPLK2 was transformed into BL21(DE3) competent cells. Protein expression was induced with isopropyl-β-D-thiogalactoside (IPTG), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed to characterize the expression and molecular weight of the rEgPLK2 protein. The purified rEgPLK2 protein was thoroughly emulsified with Freund’s complete adjuvant at a 1 : 1 volume ratio. Two New Zealand white rabbits were immunized with multipoint subcutaneous injection on the back at a dose of 300 μg per rabbit for primary immunization. For booster immunizations, the protein was emulsified with Freund’s incomplete adjuvant at a 1 : 1 volume ratio and administered on days 14, 28, and 42 after the primary immunization at a dose of 150 μg per rabbit. Serum was sampled from the rabbit ear vein on day 7 after the final immunization to yield anti-rEgPLK2 polyclonal antibodies. Antibody titer was determined by indirect enzyme-linked immunosorbent assay (ELISA), and antibody specificity was verified by Western blotting. The tissue localization of the EgPLK2 protein was detected in E. granulosus protoscoleces and adult worms using immunofluorescence assay (IFA). Eighteen 6- to 8-week-old female SPF-grade BALB/c mice were randomly divided into three groups, including the blank control group, rEgPLK2-ISA immunization group, and PBS-ISA adjuvant control group, of 6 mice each group. Mice in the rEgPLK2-ISA immunization group and PBSISA group received three primary immunizations via intramuscular injection, and animals in the rEgPLK2-ISA immunization group was inoculated with immunogens prepared by emulsifying rEgPLK2 protein with ISA 201 adjuvant at a 1 : 1 volume ratio (6 μg per mouse), while mice in the PBS-ISA adjuvant control group received an equal volume of PBS emulsified with ISA adjuvant at a 1 : 1 volume ratio. A fourth booster immunization was administered via intraperitoneal injection. Mice in the rEgPLK2-ISA immunization group received a booster immunization with 8 μg of rEgPLK2 protein per mouse, and animals in the PBS-ISA group received an equal volume of PBS, with immunizations given at 2-week intervals. Mice in the blank control group were given no treatment, and housed under standard conditions. Tail vein blood was collected from all mice 7 days after the final immunization, and levels of specific anti-rEgPLK2 IgG antibody and its subclasses (IgG1, IgG2a, IgG2b, IgG3) were measured by indirect ELISA. E. granulosus infection was modelled in mice through injection with 1 000 E. granulosus protoscoleces via intrahepatic portal vein in the rEgPLK2-ISA immunization group and PBS-ISA adjuvant control group 2 weeks after the last immunization. All mice were sacrificed and dissected. The number of cysts was counted in mouse livers, and the cyst reduction rate was calculated. Liver tissues were processed for paraffin sectioning and stained with hematoxylin and eosin (HE), and histopathological changes were examined under a light microscope. Results Sequence analysis revealed that EgPLK2 shared a high amino acid sequence homology with E. multilocularis PLK2 (EmPLK2) and contained the typical domains of the Polo-like kinase family, including the serine/threonine protein kinase catalytic domain (STKc) and Polo-box. The IPTG-induced rEgPLK2 protein was mainly expressed in the form of inclusion bodies, and the purified rEgPLK2 protein showed a relative molecular mass of approximately 70 kDa. The prepared rabbit anti-rEgPLK2 polyclonal antibody had a titer of 1 : 256 000, and Western blotting assay showed that this anti-body specifically recognized the rEgPLK2 protein with a relative molecular mass of approximately 70 kDa. Immunofluorescence assay showed that the EgPLK2 protein was localized in the excretory bladder and rostellum of E. granulosus protoscoleces, as well as the tegument, suckers, and inter-proglottid junctions of adult worms. Immunoprotective assay showed that the serum levels of specific anti-rEgPLK2 IgG, IgG1, IgG2a, and IgG2b antibodies were 2.92 ± 0.49, 0.33 ± 0.10, 0.31 (0.36), and 3.12 (1.73) in mice in the rEgPLK2-ISA immunization group, which were all significantly higher than those in the PBS-ISA adjuvant control group (0.14 ± 0.04, 0.07 ± 0.01, 0.12 ± 0.04, and 0.11 ± 0.04, respectively) (t = 19.28 and 8.46, Z = 3.75 and 4.15; all P values < 0.001); however, there was no significant difference in the serum anti-IgG3 antibody level between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group [0.07 (0.01) vs. 0.073 (0.07); Z = 0.69, P > 0.05)]. In the mouse model of E. granulosus infections, the area of hepatic lesions was reduced and the inflammatory infiltration was alleviated in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and the number of hepatic cysts was higher in the PBS-ISA adjuvant control group than in the rEgPLK2-ISA immunization group [8.00 (2.00) vs. 1.00 (0.75); Z = −2.93, P < 0.01], with a cyst reduction rate of 80.40%. Indirect ELISA assay measured higher serum levels of specific anti-rEgPLK2 IgG (3.28 ± 0.48 vs. 0.11 ± 0.04; t = 15.86, P < 0.01), IgG1 (0.29 ± 0.02 vs. 0.09 ± 0.01; t = 15.67, P < 0.01), IgG2a [3.71 (1.09) vs. 0.08 (0.03); Z = 2.88, P < 0.01], and IgG2b antibodies [3.34 (1.01) vs. 0.08 (0.03); Z = 2.88, P < 0.01] in the rEgPLK2-ISA immunization group than in the PBS-ISA adjuvant control group, and there was no significant difference in the serum level of the specific anti-rEgPLK2 IgG3 antibody between the rEgPLK2-ISA immunization group and the PBS-ISA adjuvant control group (0.07 ± 0.01 vs. 0.07 ± 0.01; t = 1.29, P > 0.05). Conclusions The prokaryotic expression system has been successfully constructed for the EgPLK2 gene and the anti-rEgPLK2 polyclonal antibody has been obtained. The rEgPLK2 protein exhibits a high immunogenicity, and is effective to protect against E. granulosus infection, and inhibits cyst development, which is a promising candidate vaccine target against cystic echinococcosis.

Objective To explore the effect of the combined application of ciprofol and remifentanil in painless fiberoptic bronchoscopy on the quality of recovery and inflammatory response.Methods 90 patients who underwent painless fiberoptic bronchoscopy from October 2023 to October 2024 were selected and randomly divided into the ciprofol combined with remifentanil group(observation group)and the propofol combined with remifentanil group(control group),with 45 cases in each group.The changes of hemodynamic indicators and BIS of the two groups of patients were compared at the time of entering the room(T0),immediately after the laryngeal mask was placed(T1),when the endoscope entered the tracheal protrusion(T2),at the end of the examination(T3),and 5 min after the laryngeal mask was removed(T4).The scores of the quality of recovery-40(QoR-40)before and after fiberoptic bronchoscopy,the levels of serum inflammatory factors,the time of fiberoptic bronchoscopy,the time to complete the instructed actions and the occurrence of adverse reactions were compared between the two groups of patients.Results The expression levels of serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)at time point T4 in the observation group were significantly lower than those in the control group,and the differences were statistically significant(P＜0.05).The comparison of BIS at T0,T1,T2,T3 and T4 time points between the two groups of patients showed no statistically significant differences(P＞0.05).The mean arterial pressure(MAP)at T,time point in the control group was significantly lower than that in the observation group,and the difference was statistically significant(P＜0.05).There was no statistically significant difference in the comparison of fiberoptic bronchoscopy examination time and the time to complete the instructed actions between the two groups of patients(P＞0.05).There was no statistically significant difference in the QoR-40 score at T0 time point between the two groups of patients(P＞0.05).At T4 time point,the comfort level,emotional state and total score of QoR-40 in the observation group were significantly higher than those in the control group,and the differences were statistically significant(P＜0.05).No serious adverse reactions occurred in either group.The incidence of hypotension in the control group was 22.2％,significantly higher than 4.4％in the observation group,and the difference was statistically significant(P＜0.05).The symptoms were relieved after fluid replacement and appropriate use of vasoactive drugs.The incidence of injection pain in the observation group was significantly lower than that in the control group,and the difference was statistically significant(P＜0.05).Conclusion The combined application of ciprofol and remifentanil in painless fiberoptic bronchoscopy is safe and effective.It has good sedative and analgesic effects,stable hemodynamics in patients,no obvious injection pain,can effectively reduce the level of inflammation cytokines,improve the quality of early recovery and patient comfort.It is worth applying in clinical practice.

Objective To explore the effect of the combined application of ciprofol and remifentanil in painless fiberoptic bronchoscopy on the quality of recovery and inflammatory response.Methods 90 patients who underwent painless fiberoptic bronchoscopy from October 2023 to October 2024 were selected and randomly divided into the ciprofol combined with remifentanil group(observation group)and the propofol combined with remifentanil group(control group),with 45 cases in each group.The changes of hemodynamic indicators and BIS of the two groups of patients were compared at the time of entering the room(T0),immediately after the laryngeal mask was placed(T1),when the endoscope entered the tracheal protrusion(T2),at the end of the examination(T3),and 5 min after the laryngeal mask was removed(T4).The scores of the quality of recovery-40(QoR-40)before and after fiberoptic bronchoscopy,the levels of serum inflammatory factors,the time of fiberoptic bronchoscopy,the time to complete the instructed actions and the occurrence of adverse reactions were compared between the two groups of patients.Results The expression levels of serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)at time point T4 in the observation group were significantly lower than those in the control group,and the differences were statistically significant(P＜0.05).The comparison of BIS at T0,T1,T2,T3 and T4 time points between the two groups of patients showed no statistically significant differences(P＞0.05).The mean arterial pressure(MAP)at T,time point in the control group was significantly lower than that in the observation group,and the difference was statistically significant(P＜0.05).There was no statistically significant difference in the comparison of fiberoptic bronchoscopy examination time and the time to complete the instructed actions between the two groups of patients(P＞0.05).There was no statistically significant difference in the QoR-40 score at T0 time point between the two groups of patients(P＞0.05).At T4 time point,the comfort level,emotional state and total score of QoR-40 in the observation group were significantly higher than those in the control group,and the differences were statistically significant(P＜0.05).No serious adverse reactions occurred in either group.The incidence of hypotension in the control group was 22.2％,significantly higher than 4.4％in the observation group,and the difference was statistically significant(P＜0.05).The symptoms were relieved after fluid replacement and appropriate use of vasoactive drugs.The incidence of injection pain in the observation group was significantly lower than that in the control group,and the difference was statistically significant(P＜0.05).Conclusion The combined application of ciprofol and remifentanil in painless fiberoptic bronchoscopy is safe and effective.It has good sedative and analgesic effects,stable hemodynamics in patients,no obvious injection pain,can effectively reduce the level of inflammation cytokines,improve the quality of early recovery and patient comfort.It is worth applying in clinical practice.

OBJECTIVE：To study the relationship of polymorphism of clopidogrel absorption and metabolism related genes CYP2C19（* 2，* 3，* 17），ABCB1 C3435T and PON1 Q192R in patients with coronary heart disease in Xinjiang Uygur Autonomous Region ，and to explore the characteristics of population and combined diseases. METHODS ：A total of 1 126 patients with coronary heart disease who underwent clopidogrel absorption and metabolism related gene testing during hospitalization in the First Affiliated Hospital of Xinjiang Medical University from January 2016 to June 2020 were included as the study subjects. The gender，age，body mass index （BMI），nationality and the proportion of combined with hypertension and diabetes were compared among different CYP2C19 metabolic phenotypes and ABCB1 C3435T and PON1 Q192R genotypes. RESULTS ：Among 1 126 patients，1 126 had CYP2C19 * 2，* 3 and * 17 genotypes，1 109 had ABCB1 C3435T genotype and 1 123 had PON1 Q192R genotype. The distribution of each genotype was in line with Hardy-Weinberg balance （P＞0.05）. There were 66（5.86％）， com 459（40.76％），476（42.27％） and 125（11.10％）patients with CYP2C19 ultra-rapid metabolizer （UM）， extensive metabolizer（EM），intermediate metabolizer （IM）and poor metabolizer（PM），respectively. The proportion of patients with UM metabolism phenotype with BMI ＞24 was significantly higher than those of patients with IM and PM metabolism phenotypes （P＜0.05）. The proportion of Han nationality patients with UM metabolic phenotype was significantly lower than those of patients with EM ，IM and PM metabolic phenotypes （P＜0.05）；the proportion of Uygur nationality was significantly higher than that of patients with EM ，IM and PM metabolic phenotypes （P＜ 0.05）. There were 355，538 and 216 patients with ABCB1 C3435T wild-type（CC），heterozygous（CT）and mutant homozygous （TT）genotypes，respectively；the proportion of Han nationality in TT genotype patients was significantly lower than that in CC and CT genotype patients （P＜0.05），and the proportion of Uygur nationality was significantly higher than that in CC and CT genotype patients （P＜0.05）；the proportion of TT genotype patients with diabetes was significantly higher than that of patients with CT genotype （P＜0.05）. There were 365，519 and 239 patients with PON1 Q192R wild-type（GG），heterozygous（GA）and mutant homozygous （AA），respectively；the proportion of Han nationality in AA genotype patients was significantly lower than that in GG and GA genotype patients （P＜0.05），and the proportion of Uygur nationality was significantly higher than that of GG and GA genotype patients （P＜0.05）；the proportion of Han nationality and BMI ≤24 in patients with AA genotype were significantly lower than those with GA genotype （P＜0.05），and the proportion of Uygur nationality ，BMI＞24 and hypertension were significantly higher than those in GA genotype patients （P＜0.05）. CONCLUSIONS ：There are significant nationality differences among patients with different CYP2C19 metabolic phenotypes and ABCB1 C3435T and PON1 Q192R genotypes. In addition，patients with BMI ＞24 account for high proportion among CYP2C19 UM metabolism genotype ；patients with diabetes account for high proportion among ABCB1 C3435T TT genotype ；patients with BMI ＞24 and hypertension account for high proportion among PON1 Q192R AA genotype.

This study aimed to evaluate the effect of sanguinarine on biomechanical properties of rat airway smooth muscle cells (rASMCs) including stiffness, traction force and cytoskeletal stress fiber organization. To do so, rASMCs cultured were treated with sanguinarine solution at different concentrations (0.005~5 μmol/L) for 12 h, 24 h, 36 h, and 48 h, respectively. Subsequently, the cells were tested for their viability, stiffness, traction force, migration and microfilament distribution by using methylthiazolyldiphenyl-tetrazolium bromide assay, optical magnetic twisting cytometry, Fourier transform traction microscopy, scratch wound healing method, and immunofluorescence microscopy, respectively. The results showed that at concentration below 0.5 μmol/L sanguinarine had no effect on cell viability, but caused dose and time dependent effect on cell biomechanics. Specifically, rASMCs treated with sanguinarine at 0.05 μmol/L and 0.5 μmol/L for 12 and 24 h exhibited significant reduction in stiffness, traction force and migration speed, together with disorganization of the cytoskeletal stress fibers. Considering the essential role of airway smooth muscle cells (ASMCs) biomechanics in the airway hyperresponsiveness (AHR) of asthma, these findings suggest that sanguinarine may ameliorate AHR via alteration of ASMCs biomechanical properties, thus providing a novel approach for asthma drug development.

Objective To investigate the expression and effect of miR-22-3p in non-small cell lung cancer (NSCLC). Methods The miR-22-3p expression level in seventy-six NSCLC tissues and para-cancer tissues was detected by qRT-PCR. The relationship between the expression of miR-22-3p and gender,age,tumor size,histolo-gy grade,pathological type and lymph node metastasis was analyzed. The function of miR-22-3p on the prolifera-tion of NSCLC cells was tested by growth curve assay. Target genes of miR-22-3p were predicted by online software Targetscan. Luciferase reporter assay and qRT-PCR was used to certificate the prediction. Results The expression of miR-22-3p was increased in NSCLC tissues than the para-cancer tissues and was correlated to lymph node metas-tasis. Overexpression of miR-22-3p could suppress the proliferation of A549 cells. Astrocyte-Elevated Gene-1(AEG-1) was predicted to be a target of miR-22-3p. MiR-22-3p was revealed to bind to AEG-13′UTR by luciferase report-er assay. Overexpression of miR-22-3p could inhibit the expression of AEG-1 in A549 cells. Suppression of miR-22-3p could increase AEG-1 expression. Conclusion MiR-22-3p could inhibit the proliferation of NSCLC by tar-geting AEG-1.

Objective To explore the significance of detecting miR-145 hypermethylation in non-small cell lung cancer (NSCLC). Methods Seventy-five NSCLC tissues and adjacent tissues was collected from May 2012 to January 2015. The methylation status of miR-145 promoter was tested by bisulfite sequencing PCR (BSP) and the expression of miR-145 evaluated by ISH. The correlation between miR-145 methylation and clinical parameters , and the relationship between miR-145 methylation and expression were statistically analyzed. Results The hypermethy-lation ratio of miR-145 in NSCLC tissues was higher than that in the adjacent tissues. Hypermethylation of miR-145 was correlated with NSCLC differentiation stage, but not with gender and age. In NSCLC, miR-145 hypermethyla-tion was negatively related with its expression. Conclusion MiR-145 hypermethylation is closely related with dif-ferentiation stage in NSCLC tissues. MiR-145 hypermethylation may be a potential biomarker for estimating NSCLC differentiation stage.

To evaluate the ultrasound imaging characteristics and diagnostic criteria for acute abdominal Meckel's di- verticulum disease (MD), we retrospectively analyzed the ultrasonic characteristics, clinical data of 58 cases of pathologically proved MD from January 2009 to May 2012. We found that among all the 58 patients, 21 patients were diagnosed with the preoperative clinical diagnosis of MD. Fourteen cases of MD inflammation with acute appendicitis were evaluated by pathological examinations after the surgery. We also found 4 cases of MD with perforation, 15 cases of MD with intussusceptions, 14 cases MD with intestinal obstruction, 5 cases of MD secondary to intestinal obstructionor intestinal necrosis, and 5 cases of MD without any obvious complications. Emergency ultrasound examinations revealed 8 cases of simple MD, 1 case of MD with intussusceptions, 9 cases of MD with acute appendicitis, 12 cases of MD with intestinal obstruction, 2 cases of MD with intussusceptions and intestinal obstruction, 1 case of MD with omphalocele and 1 case of MD with abdominal abscess. The emergency sonographic findings suggested that MD was relatively fixed bowel or thick-walled cystic mass, with one end connected to small intestine, and the other end connected to the blind side, at the periumbilicus region or at the lower right abdomen. A conclusion could be drawn that MD is difficult to be detected by ultrasound (detection rate was about 15. 5%), and MD with complications such as intussusceptions, intestinal obstruction, acute appendicitis can usually be more easily detected (detection rates were 24.1%, 24.1% and 15.5%, respectively). Sonography is a simple, effective way to make diagnosis and differential diagnosis of MD with different acute abdomen symptoms from other disease.
Acute Disease ; Appendicitis ; pathology ; Diagnosis, Differential ; Humans ; Inflammation ; Intestine, Small ; pathology ; surgery ; Meckel Diverticulum ; diagnostic imaging ; surgery ; Retrospective Studies ; Ultrasonography

The aim of the present study was to investigate the diagnostic value of the contrast-enhanced ultrasonography for gastrointestinal stromal tumors (GISTs). The conventional transabdominal ultrasonography and contrast-enhanced ultrasonography using an intraluminal contrast agent were performed in 66 cases. The different diagnostic value between conventional transabdominal ultrasonography and contrast-enhanced hydrocolon ultrasonography was compared, including the depiction ratio and location accuracy. The morpholpgical features of GISTs were observed. The sensitivity of the contrast-enhanced hydrocolon ultrasonography in the depiction of the GISTs (57/66, 86.4%) was higher than that of the conventional transabdominal ultrasonography(33/66, 50%, P<0. 05). Furthermore, the accuracy of the contrast-enhanced ultrasonography in the location of the GISTs (52/57, 91. 2%) was higher than that of the conventional transabdominal ultrasonography (13/33, 39. 4%, P<0. 05). The GISTs with higher or lower risk have different ultrasonic features. The contrast-enhanced ultrasonography was a more valuable diagnostic method for GIST than the conventional transabdominal ultrasonography.
Adult ; Aged ; Contrast Media ; administration & dosage ; Endosonography ; methods ; Female ; Gastrointestinal Neoplasms ; diagnostic imaging ; Gastrointestinal Stromal Tumors ; diagnostic imaging ; Humans ; Male ; Middle Aged

Objective To analyze the pathologic and biology features and prognosis of HER-2,ER and PR negative breast cancer(TNBC).Methods Clinic pathologic data of 329 breast cancer patients was retrospectively analyzed. The expression of HER-2, ER, PR and P53 was determined by immunohistochemistry. The patients were divided into three groups, including TN group [ HER-2 (-), ER (-), PR (-)], HER-2 group [HER-2(+),ER(-),PR(-)] and HR group [ER(+),PR(-) or (+),HER-2(-) or (+) ].The pathology and biology features and P53 masculine expression of the three groups were compared.The 5-year overall survival and disease free survival were analyzed by Kaplan-Meier method.Results Of the 329 patients, 20.97% (69/329) was TN group, 34.04% (112/329)was HER-2 group, and 44.98% (148/329) was HR group.The percentage of with lymph node metastasis in TN group 55.07% (38/69) was higher than that in HER-2 group 42.86% (48/112) and HR group 43.24% (64/148) (χ2 = 12.57, P ＜ 0.05).The rates of P53 positive, operation recurrence and metastasis in TN group were 44.93%, 27.54% and 20.29%, which were higher than that in HER-2 group (20.54% ,16.07%,16.96%) and HR group (18.24%, 12.84%, 10.81%) (χ2=12.23, 8.36, P ＜0.05).The ratio of tumor ≥5cm, Ⅲ stages, Ⅲ grade and soakage canula cancer among three groups had statistical difference (χ2 = 7.25,8.79,9.23,8.48, P ＜ 0.05).The 5-year overall survival in three groups were 75.36%,82.14% ,85.14% and disease free survival were 68.12% ,78.57% ,82.43% (χ2 =8.52, P ＜0.05).Conclusion The pathology and biology traits of TNBC were high rate of P53 (+) and recurrence and lymph node metastasis.The most important factor for poor prognosis of TNBC was the low rate of disease free survival for 5 years.