1.Effect of buccal acupuncture on analgesia after tonsilloadenoidectomy in pediatric patients
Yaying HUANG ; Jiayi YANG ; Jiehui FANG ; Haoxiang KE ; Yingyi XU ; Bilian LI ; Junxiang HUANG ; Xingrong SONG ; Tingting YU
Chinese Journal of Anesthesiology 2024;44(10):1235-1238
Objective:To evaluate the effect of buccal acupuncture on analgesia after tonsilloadenoidectomy in pediatric patients.Methods:This was a randomized controlled study. One hundred and twenty-six American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ pediatric patients, aged 3-12 yr, weighing 12-34 kg, with body mass index <30 kg/m 2, undergoing elective tonsilloadenoidectomy with general anesthesia, were divided into 2 groups ( n=63 each) by the random number table method: buccal acupuncture group (group B) and control group (group C). All pediatric patients received the same anesthesia induction and intraoperative anesthesia maintenance. The concentration of sevoflurane was adjusted to keep the fluctuation amplitude of vital sign parameters within 20% of the baseline value. After surgery, the drug was immediately stopped and the children were transferred to the postanesthesia care unit for resuscitation under general anesthesia. In group B, the bilateral neck points, upper neck points, hologram points on the head and Zhongjiao points were selected before removal of the tracheal catheter, and disposable acupuncture needles were inserted directly into the acupoints and remained for 20-30 min. Group C received no buccal acupuncture. The pain Assessment Scale (FLACC) was used to assess the severity of postoperative pain. The postoperative agitation score was evaluated by Aono four-point rating method to evaluate the occurrence of agitation. The effective pressing times of patient-controlled analgesia, rescue analgesia and occurrence of nausea and vomiting within 48 h after operation were recorded. The occurrence of bleeding, infection and broken needle at acupuncture sites was recorded. Results:Compared with group C, the effective pressing times of patient-controlled analgesia and incidence of nausea and vomiting were significantly decreased in group B ( P<0.05). There was no significant difference in the rate of rescue analgesia and incidence of postoperative agitation between the two groups ( P>0.05). No infection or broken needle was found at acupuncture sites after buccal acupuncture, only 2 cases had slight bleeding at the puncture site, and there was no abnormality after pressing in group B. Conclusions:Buccal acupuncture can enhance the analgesic effect after tonsilloadenoidectomy in pediatric patients.
2.Molecular mechanisms underlying the inflammatory response induced by Cutibacterium acnes biofilms in keratinocytes
Lu PEI ; Nana ZHENG ; Rong ZENG ; Yuanyuan XIE ; Haoxiang XU ; Zhimin DUAN ; Yuzhen LIU ; Min LI
Chinese Journal of Dermatology 2024;57(4):302-308
Objective:To investigate molecular mechanisms underlying the inflammatory response induced by Cutibacterium acnes ( C. acnes) biofilms in human primary keratinocytes. Methods:A C. acnes biofilm model was established in vitro, and confocal fluorescence microscopy was performed to examine its three-dimensional structure. The cultured human primary keratinocytes were divided into 3 groups: a dimethyl sulfoxide (DMSO) control group (treated with 0.01% DMSO alone), a C. acnes suspension group (co-incubated with C. acnes suspensions), and a C. acnes biofilm group (co-incubated with C. acnes biofilms). Real-time fluorescence-based quantitative PCR (RT-qPCR) was performed to determine the relative mRNA expression of interleukin (IL) -6, IL-8, and tumor necrosis factor (TNF) -α in the groups after 6-hour culture, enzyme-linked immunosorbent assay to detect the free protein levels of IL-6, IL-8, and TNF-α in the groups after 24-hour culture, and Western blot analysis to determine the protein expression of Toll-like receptor 2 (TLR2) in keratinocytes. In addition, some human primary keratinocytes were pretreated with key molecular blockers targeting the TLR2/mitogen-activated protein kinase (MAPK) /nuclear factor (NF) -κB signaling pathway (C29, ST2825, BAY11-7082, SB203580, U0126-EtOH), and then co-incubated with C. acnes biofilms; the DMSO control group and the C. acnes biofilm group receiving no pretreatment were simultaneously set as negative and positive controls, respectively. The mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were then detected in the above groups. One-way analysis of variance was used for comparisons among multiple groups, and the Bonferroni method was used for multiple comparisons. Results:Confocal fluorescence microscopy demonstrated a three-dimensional C. acnes biofilm structure resembling a lawn, and the biofilm grew well. RT-qPCR and ELISA showed significant differences in the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α among the C. acnes biofilm group, C. acnes suspension group and DMSO control group (mRNA: F = 89.70, 312.17, 46.09, respectively, all P < 0.001; free protein: F = 886.12, 634.25, 307.01, respectively, all P < 0.001) ; in detail, the mRNA and free protein expression levels of IL-6, IL-8, and TNF-α were significantly higher in the C. acnes biofilm group than in the C. acnes suspension group and DMSO control group (all P < 0.001) ; the C. acnes suspension group showed significantly increased expression levels of IL-6 mRNA and TNF-α free protein compared with the DMSO control group ( P < 0.001, = 0.003, respectively), while there were no significant differences in the expression of IL-6 free protein, TNF-α mRNA, or IL-8 mRNA and free protein between the 2 groups (all P > 0.05). Western blot analysis showed that the TLR2 protein expression was significantly higher in the C. acnes suspension group and C. acnes biofilm group than in the DMSO control group. After the pretreatment with molecular blockers targeting the MAPK/NF-κB signaling pathway and co-incubation with C. acnes biofilms, the mRNA and free protein expression levels of IL-6, IL-8 and TNF-α were all significantly lower in the C29 group, ST2825 group, BAY11-7082 group, SB203580 group, U0126-EtOH group, as well as in the DMSO control group compared with the C. acnes biofilm group (all P < 0.05) . Conclusion:The C. acnes biofilms exhibited a strong ability to induce inflammatory responses in human keratinocytes, possibly through the activation of the TLR2/MAPK/NF-κB signaling pathway.
3.Discussing the Staged Treatment of Progressive Fibrosing Interstitial Lung Disease Based on the Theory of "Opening-Closing-Pivoting"
Haoxiang ZHANG ; Jin'e WANG ; Xiao XIA ; Yanyi LIU ; Yujin DING ; Yingzi TIAN ; Kun XIA ; Guangxi LI
Journal of Traditional Chinese Medicine 2024;65(18):1882-1887
Progressive fibrosing interstitial lung disease (PF-ILD) has a complex etiology, and is classified as lung impediment stage, impediment-atrophy combination stage, and lung atrophy stage according to the different clinical manifestations during the progression of disease. Based on the theory of opening-closing-pivoting to analyse the characteristics of yin and yang disease mechanism and the idea of prescriptions in the three stages. For lung impediment stage, main as three-Yang fail to keep inside, disharmony between Ying qi (营气) and Wei qi (卫气), shaoyin impairment, treatment should use Mahuang (Ephedra sinica) and Guizhi (Neolitsea cassia) flexibly to form a formula, or choose pungent-dispersing formulas like Baidu Powder (败毒散) to move qi and save yang, and diffuse and disperse impediment pathogen, meanwhile combining saving-shaoyin medicinals like Fuzi (Aconitum carmichaelii) and Shudihuang (Radix Rehmanniae Praeparata) to reinforce healthy qi and dispel pathogen; for impediment-atrophy combination stage, rooted as yangming impairment and progressed by over-movement of qi, treatment should use Mahuang Shengma Decoction (麻黄升麻汤) to resolve and decrease over-activities, emphasis on both opening and closing, and improve impediment and atrophy; for lung atrophy stage with three-Yin in a bad condition simultaneously and poor prognosis, treatment should use modified Jinshui Liujun Decoction (金水六君煎) to consolidate qi and save yin, disperse phlegm and stasis, to improve the quality of life for patients with PF-ILD.
4.Mendelian randomization and bioinformatics analysis of the disulfidoptosis core gene SLC7A11 in clear cell renal cell carcinoma
Zifeng LI ; Bohong CHEN ; Haoxiang HUANG ; Cong FENG ; Jin ZENG ; Wei CHEN ; Dapeng WU
Journal of Modern Urology 2024;29(5):459-465,475
Objective To investigate the role of solute carrier family 7 member 11(SLC7A11)in the pathogenesis and progression of clear cell renal cell carcinoma(ccRCC)and its prognostic significance.Methods Mendelian randomization analysis was employed to identify genes causally associated with the risk of ccRCC.The expression patterns and prognostic relevance of SLC7A11 were assessed using RNA sequencing data and clinical information obtained from the UCSC Xcna pan-cancer cohort.Gene set enrichment analysis(GSEA)was conducted using data from The Cancer Genome Atlas Kidney Renal Clear Cell Carcinoma(TCGA-KIRC)dataset(training set).A prognostic model based on SLC7A11 was then developed using stepwise Cox regression and validated externally in the E-MTAB-1980 cohort(validation set).Results Elevated level of SLC7A11 was associated with an increased risk of ccRCC(HR=1.27,95%CI:1.15-1.40,P<0.001).SLC7A11 was overexpressed in various tumors and correlated with higher T stage and poorer survival(P<0.05).GSEA demonstrated that SLC7A11 was enriched in pathways related to proliferation and metastasis,including E2F and epithelial-to-mesenchymal transition signaling pathways.Moreover,the SLC7A11 prognostic model exhibited robust predictive performance in both the training set(1-,3-,and 5-year AUC=0.78,0.73,0.71,respectively)and the external validation set(1-,3-,and 5-year AUC=0.70,0.71,0.72,respectively).Conclusion SLC7A11 can be a potential biomarker and therapeutic target for ccRCC,offering novel perspectives for precision medicine.
5.CHESS endoscopic ruler in objective measurement of diameter of esophageal varices in liver cirrhosis and portal hypertension: a prospective multicenter study
Shengjuan HU ; Jianping HU ; Shaoqi YANG ; Xiaoguo LI ; Yanhong DENG ; Ruichun SHI ; Xiaoqin LI ; Hailong QI ; Qian SHEN ; Fang HE ; Jun ZHU ; Bin MA ; Xiaobing YU ; Jianyang GUO ; Yuehua YU ; Haijiang YONG ; Wentun YAO ; Ting YE ; Hua WANG ; Wenfu DONG ; Jianguo LIU ; Qiang WEI ; Jing TIAN ; Haoxiang HE ; Changhui HE ; Yifei HUANG ; Yang BU ; Xiaolong QI
Chinese Journal of Digestion 2023;43(3):193-198
Objective:To investigate the safety and feasibility of the CHESS endoscpic ruler (CHESS ruler), and the consistency between the measured values and the interpretation values by endoscopic physician experience.Methods:From January 2021 to January 2022, a total of 105 liver cirrhosis patients with portal hypertension were prospectively enrolled from General Hospital, Xixia Branch Hospital, Ningnan Hospital of People′s Hospital of Ningxia Hui Autonomous Region (29 cases), and the First People′s Hospital of Yinchuan (25 cases), General Hospital of Ningxia Medical University (18 cases), Wuzhong People′s Hospital (10 cases), the Fifth People′s Hospital of Ningxia Hui Autonomous Region (10 cases), Shizuishan Second People′s Hospital (6 cases), Yinchuan Second People′s Hospital (5 cases), and Zhongwei People′s Hospital (2 cases) 8 hospitals. The clinical characteristics of all the patients, including gender, age, nationality, etiolog of liver cirrhosis, and Child-Pugh classification of liver function were recorded. A big gastroesophageal varices was defined as diameter of varices ≥5 mm. Endoscopist (associated chief physician) performed gastroscopy according to the routine gastroscopy procedures, and the diameter of the biggest esophageal varices was measured by experience and images were collected, and then objective measurement was with the CHESS ruler and images were collected. The diameter of esophageal varices of 10 randomly selected patients (random number table method) was determined by 6 endoscopists (attending physician or associated chief physician) with experience or measured by CHESS ruler. Kappa test was used to test the consistency in the diameter of esophageal varices between measured values by CHESS ruler and the interpretation values by endoscopic physician experience.Results:Among 105 liver cirrhosis patients with portal hypertension, male 65 cases and female 40 cases, aged (54.8±12.2) years old, Han nationality 82 cases, Hui nationality 21 cases and Mongolian nationality 2 cases. The etiology of liver cirrhosis included chronic hepatitis B (79 cases), alcoholic liver disease (7 cases), autoimmune hepatitis (7 cases), chronic hepatitis C (2 cases), and other etiology (10 cases). Liver function of 32 cases was Child-Pugh A, Child-Pugh B 57 cases, and Child-Pugh C 16 cases. All 105 liver cirrhosis patients with cirrhotic portal hypertension were successfully measured the diameter of gastroesophageal varices by CHESS ruler, and the success rate of application of CHESS ruler was 100.0% (105/105). The procedure time from the CHESS ruler into the body to the exit of the body after measurement was (3.50±2.55) min. No complications happened in all the patients during measurement. Among 105 liver cirrhosis patients with cirrhotic portal hypertension, 96 cases (91.4%) were recognized as big gastroesophageal varices by the endoscopists. Totally 93 cases (88.6%) were considered as big gastroesophageal varices by CHESS ruler. Eight cases were recognized as big gastroesophageal varices by the endoscopist, however not by the CHESS ruler; 5 cases were recognized as big gastroesophageal varices by the CHESS ruler, but not by the endoscopists; 4 cases were not recognized as big gastroesophageal varices both by the endoscopists and CHESS ruler; 88 cases were recognized as big gastroesophageal varices both by the endoscopists and CHESS ruler. The missed diagnostic rate of big gastroesophageal varices by the endoscopists experience was 5.4% (5/93), and the Kappa value of consistency coefficient between the measurement by the CHESS ruler and the interpretation by endoscopists experience was 0.31 (95% confidence interval 0.03 to 0.60). The overall Kappa value of consistency coefficient by 6 endoscopists measured by CHESS ruler in big gastroesophageal varices diagnosis was 0.77 (95% confidence interval 0.61 to 0.93).Conclusion:As an objective measurement tool, CHESS ruler can make up for the deficiency of subjective judgment by endoscopists, accurately measure the diameter of gastroesophageal varices, and is highly feasible and safe.
6.Development of Human Vital Signs and Body Posture Monitoring and Positioning Alarm Systems.
Haoxiang TANG ; Jia XU ; Ruijing SHE ; Dongni NING ; Yushun GONG ; Yongqin LI ; Liang WEI
Chinese Journal of Medical Instrumentation 2023;47(6):617-623
In view of the high incidence of malignant diseases such as malignant arrhythmias in the elderly population, accidental injuries such as falls, and the problem of no witnesses when danger occurs, the study developed a human vital signs and body posture monitoring and positioning alarm system. Through the collection and analysis of electrocardiogram (ECG), respiration (RESP) and acceleration (ACC) signals, the system monitors human vital signs and body posture in real time, automatically judges critical states such as malignant arrhythmias and accidental falls on the local device side, and then issues alarm information, opens the positioning function, and uploads physiological information and patient location information through 4G communication. Experiments have shown that the system can accurately determine the occurrence of ventricular fibrillation and falls, and issue position and alarm information.
Humans
;
Aged
;
Arrhythmias, Cardiac/diagnosis*
;
Ventricular Fibrillation
;
Electrocardiography
;
Accidental Falls
;
Vital Signs
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Posture
;
Monitoring, Physiologic
7.Inhibitory effect of deoxyribonucleaseⅠ against Cutibacterium acnes biofilms
Meng ZHOU ; Nana ZHENG ; Rong ZENG ; Haoxiang XU ; Zhimin DUAN ; Yuzhen LIU ; Min LI
Chinese Journal of Dermatology 2023;56(10):920-924
Objective:To investigate the inhibitory effect of deoxyribonuclease Ⅰ (DNaseⅠ) on Cutibacterium acnes biofilms. Methods:Cutibacterium acnes biofilms were constructed, and then were divided into 4 groups (negative control group, 5, 10 and 20 U/ml DNase Ⅰ groups) to be treated with DNase Ⅰ at different concentrations of 0, 5, 10 and 20 U/ml respectively. The biofilm viability was evaluated by tetrazolium salt colorimetric assay, the biofilm content was determined by crystal violet staining-based semi-quantitative analysis, the biofilm structure was observed by confocal laser scanning microscopy, and the live/dead bacteria ratio was calculated. One-way analysis of variance was used to analyze differences between groups. Results:After the treatment with DNase Ⅰ, the biofilm viability was significantly inhibited in the 5, 10 and 20 U/ml DNaseⅠ groups (1.882 ± 0.421, 1.653 ± 0.287, 1.473 ± 0.154, respectively) compared with the negative control group (2.668 ± 0.245), and the inhibitory effect was gradually enhanced with the increase in concentrations of DNase Ⅰ ( F = 9.68, P = 0.005). Crystal violet semi-quantitative analysis showed that the biofilm content was also significantly lower in the 5, 10 and 20 U/ml DNaseⅠ groups (1.039 ± 0.003, 0.489 ± 0.079, 0.147 ± 0.034, respectively) than in the negative control group (1.359 ± 0.071), and the higher the DNase Ⅰ concentration, the lower the biofilm content ( F = 174.40, P < 0.001). Confocal laser scanning microscopy showed that the biofilm structure was destroyed in the 5, 10 and 20 U/ml DNase Ⅰ groups compared with the negative control group, and the higher the DNase Ⅰ concentration, the more severe the destruction of biofilm structure. Additionally, the live/dead bacteria ratio was significantly lower in the 5, 10 and 20 U/ml DNaseⅠ groups (2.303 ± 0.457, 1.534 ± 0.526, 1.263 ± 0.354, respectively) than in the negative control group (4.475 ± 0.146), and the ratio decreased with the increase in concentrations of DNase Ⅰ ( F = 56.75, P < 0.000 1) . Conclusion:DNase Ⅰ had a destructive effect on the structure of Cutibacterium acnes biofilms, and could inhibit their viability.
8.Effect of different incubation time of aminolevulinic acid on photodynamic inhibition of Propionibacterium acnes biofilms
Yuzhen LIU ; Rong ZENG ; Nana ZHENG ; Zhimin DUAN ; Haoxiang XU ; Qiuju WU ; Tong LIN ; Min LI
Chinese Journal of Dermatology 2022;55(3):208-212
Objective:To investigate the effect of different incubation time of aminolevulinic acid (ALA) on photodynamic inhibition of Propionibacterium acnes biofilms. Methods:Propionibacterium acnes biofilms were formed in 24-well plates with pre-placed cell slides and 96-well plates. The formation of the biofilm structure was observed by confocal laser scanning microscopy (CLSM) , and the growth activity of the biofilm was assessed by the tetrazolium salt XTT assay. The in vitro successfully constructed biofilm models were divided into 6 groups: negative control group receiving neither ALA treatment nor LED radiation, ALA group incubated with ALA alone for 30 minutes, LED group receiving LED radiation alone, ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group incubated with ALA for 15, 30 and 60 minutes respectively followed by LED radiation. After the treatment, CLSM was performed to observe the biofilm structure, as well as to determine the dead/living bacteria ratio, and XTT assay to assess the growth activity of the biofilm. Differences among groups were analyzed using one-way analysis of variance and least significant difference- t test. Results:CLSM showed that the Propionibacterium acnes biofilm model was successfully constructed in vitro. The dead/living bacteria ratios were 0.90 ± 0.16, 1.75 ± 0.19, and 2.57 ± 0.32 in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group respectively, which were significantly higher than the dead/living bacteria ratio in the negative control group (0.31 ± 0.01; t= 55.56, 138.62, 74.64, respectively, all P<0.001) ; the biofilm viability value was significantly lower in the ALA-PDT1 group, ALA-PDT2 group and ALA-PDT3 group (0.35 ± 0.02, 0.26 ± 0.02, 0.18 ± 0.01, respectively) than in the negative control group (0.43 ± 0.00; t= 35.66, 2.64, 110.96, respectively, all P < 0.001) . CLSM showed that the structure of the Propionibacterium acnes biofilm was destroyed under the action of ALA-PDT, and the destruction was aggravated with the prolongation of incubation time of ALA. Conclusion:The prolongation of incubation time of ALA can enhance the inhibitory effect of ALA-PDT on Propionibacterium acnes biofilms.
9.Clinical characteristics and surgical effects of acute calculous cholecystitis in high altitude area of Tibet
Jie SHU ; Haoxiang ZHANG ; Jianwei LI ; Lin LI ; Qingchun CHANG ; Danzeng SUOLANG ; Jifeng ZENG ; Fangfang ZHENG ; Li CAO ; Shuguo ZHENG
Chinese Journal of Digestive Surgery 2022;21(7):917-922
Objective:To investigate the clinical characteristics and surgical effects of acute calculous cholecystitis (ACC) in high altitude area of Tibet.Methods:The retrospective cohort study was conducted. The clinicopathological data of 182 ACC patients who underwent surgery in the 954th Hospital of Army from January 2016 to December 2020 were collected. There were 56 males and 126 females, aged (41±13)years. Of the 182 patients, 61 cases undergoing open cholecystec-tomy were divided into the open group, and 121 cases undergoing laparoscopic cholecystectomy (LC) were divided into the laparoscopic group. Observation indicators: (1) clinical characteristics of ACC in high altitude area; (2) surgical situations; (3) postoperative complications; (4) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect postopera-tive complications of patients up to October 2021. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measure-ment data with skewed distribution were represented as M( Q1, Q3) or M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Results:(1) Clinical characteristics of ACC in high altitude area. Of the 182 patients, cases with symptom duration as <3 days, 3 days to 1 month, >1 month and ≤12 months, >12 months were 37, 43, 57, 45, respectively. Seventy-seven of the 182 patients were combined with other diseases before surgery. (2) Surgical situations. Two cases in the open group were found common bile duct stones during the operation, and underwent choledochotomy and T-tube drainage. Nine cases in the laparoscopic group were converted to laparotomy, including 3 cases with severe abdominal adhesion and ineffective hemostasis, 6 cases with anatomical variation of Calot triangle. The conversion to laparotomy rate was 7.438%(9/121). The other patients in the open group and the laparoscopic group completed surgery successfully. The operation time, volume of intraoperative blood loss, time to postoperative first out-of-bed activities, time to postoperative first flatus, cases with indwelling drainage tube, cases with acute simple cholecystitis, acute suppurative cholecystitis, acute gangrene cholecystitis, gallbladder perforation of disease pathological type, postoperative white cell count, postoperative neutrophil percentage, duration of postoperative hospital stay were (109±42)minutes, 50(45,100)mL, (16.1±1.5)hours, (31.4±11.9)hours, 33, 25, 27, 6, 3, (6.8±1.9)×10 9/L, 72.7%±7.4%, (7.3±1.7)days for the open group. The above indicators were (98±43)minutes, 20(20,50)mL, (12.9±1.4)hours, (26.7±12.1)hours, 51, 56, 51, 9, 5, (7.1±2.4)×10 9/L, 70.5%±8.7%, (6.4±1.7)days for the laparoscopic group. There were significant differences in the volume of intraopera-tive blood loss, time to postoperative first out-of-bed activities, time to postoperative first flatus, duration of postoperative hospital stay between the two groups ( Z=?6.75, t=14.41, 2.46, 3.45, P<0.05). There was no significant difference in the operation time, cases with indwelling drainage tube, diseases pathological type, postoperative white cell count, postoperative neutrophil percentage between the two groups ( t=1.66, χ2=2.33, 0.84, t=?0.71, 1.66, P>0.05). (3) Postoperative complica-tions. Postoperative complications occurred in 7 of the 61 patients in the open group and 5 of the 121 patients in the laparoscopic group. There was no significant difference in the postoperative complications between the two groups ( χ2=2.46, P>0.05). (4) Follow-up. Of the 182 patients, 115 cases including 35 cases in the open group and 80 cases in the laparoscopic group were followed up for 12(range, 3?24)months. During the follow-up, 1 case of the 35 patients in the open group had abdominal pain and jaundice, which was diagnosed as choledocholithiasis. The patient was improved after stone removal with endoscopic retrograde cholangiopancreatography. Two cases of the 35 patients in the open group had upper abdominal pain with fever and were improved after anti-infection treatment. Of the 80 patients in the laparoscopic group, 1 case had upper abdominal pain and 1 case had dyspepsia and anorexia, respectively. The two cases were improved after symptomatic treatment. Conclusions:Patients with ACC in the high altitude area of Tibet have high ratio of preoperative complications, long diseases history and high incidence rates of pyogenic perforation of the gallbladder. Patients with ACC in the high altitude area undergoing LC is safe and effective. Compared with open cholecystectomy, LC have less volume of intraoperative blood loss, faster postoperative recovery and shorter duration of postoperative hospital stay.
10.Construction of recombinant adenoviral vector overexpressing PTG
Chenxi Wang ; Xia Deng ; Zhicong Zhao ; Zhensheng Cai ; Panpan Zhang ; Lian Li ; Haoxiang Li ; Li Zhao ; Dong Wang ; Ling Yang ; Guoyue Yuan
Acta Universitatis Medicinalis Anhui 2022;57(4):558-563
Objective:
To construct and identify an overexpression recombinant adenovirus vector carrying the mouse PTG gene(NM_016854), and to lay a foundation for in-depth study of the function of PTG.
Methods:
The coding sequence of the mouse PTG gene was chemically synthesized, amplified by polymerase chain reaction(PCR), digested with restriction enzymes, and inserted into the GV314 vector(CMV-MCS-3 FLAG-SV40-EGFP) to obtain the recombinant shuttle plasmid pGV314-PTG. BamHⅠ/AgeⅠ double enzyme digestion was further carried out, and the product was transferred into linearized expression vector pDC315 to construct recombinant adenovirus Ad-PTG, which was transfected into HEK293 T cells and packaged into recombinant virus particles. After repeated amplification of several generations of HEK293 T cells, the recombinant adenovirus was purified and titer detected. Finally, PCR, Western blot and sequencing were used to verify the recombinant adenovirus.
Results:
After PCR, Western blot and sequencing, the results showed that the pGV314-CMV-MCS-3 FLAG-SV40-EGFP-PTG overexpression adenovirus vector(Ad-PTG) was successfully constructed, and the virus titer measured by end-point dilution method was 4×1010PFU/ml, Western blot and RT-qPCR showed that the protein and mRNA expression levels of PTG increased significantly.
Conclusion
The recombinant adenovirus vector carrying mouse PTG gene is successfully constructed, and the expression of PTG gene in hepatocytes is effectively up regulated.


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