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Objective:To investigate the expression of Mucosal-associated invariant T (MAIT) cells in the peripheral blood of ankylosing spondylitis (AS) patients, and to analyze its clinical significance.Methods:Sixty-four AS patients from September 2020 to September 2022 in Fujian Provincial Hospital and 60 healthy controls were enrolled. The expression of MAIT cells and CD69 +MAIT cells were detected by immunofluorescence. Spondylarthritis research consortium of Canada (SPARCC) scores of sacroiliac joint magnetic resonance imaging (MRI) were calculated and were analyzed based on clinical data. The changes of MAIT cells were observed in 7 patients with AS after treatment. t test, variance analysis and Pearson correlation analysis were used for statistical analysis. Results:The expression of MAIT cells in the peripheral of AS patients was significantly lower than that in the controls [(2.81±0.27)% and (4.46±0.86)% respectively, t=2.33, P=0.022], while CD69 +MAIT cells were significantly higher [(12.0±1.0)% and (7.8±1.2)% respectively, t=2.31, P=0.024]. The level of CD69 +MAIT cells were significantly positively correlated with erythrocyte sedimentation rate (ESR) ( r=0.39, P=0.010), C-reactive protein (CRP) ( r=0.36, P=0.012), ASDAS ( r=0.35, P=0.013) and SPARCC scores ( r=0.38, P=0.006) of AS patients. ASDAS scores and CD69 +MAIT cells obviously decreased in 7 treated AS patients ( F=7.62, P=0.007 and F=4.97, P=0.027 respectively). Conclusion:The expression of peripheral MAIT cells in AS patients is lower than that in healthy controls, but the number of the activated MAIT cells is increased. Levels of activated MAIT cells may in some extent reflect the inflammatory state and disease activity of AS, as well as inflammatory destruction of sacroiliac joint, and therapeutic effect. MAIT cells may partially participate in the inflammatory response and play a role in the pathogenesis of AS.
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Objective:To compare effect of internal fixation of ribs assisted by complete thoracoscopy and thoracotomy for flail chest.Methods:A retrospective cohort study was used to analyze the clinical data of 86 patients with flail chest treated at No.2 Hospital of Nanping City and 900th Hospital of Joint Logistics Support Force between January 2019 and December 2020, including 58 males and 28 females; aged 25-69 years [(42.9±9.5)years]. A total of 45 patients underwent internal fixation of ribs assisted by complete thoracoscopy (thoracoscopy group), and 41 patients by thoracotomy (thoracotomy group). The operation time, number of fixed ribs, intraoperative blood loss, ventilation time, postoperative length of hospital stay, hemodynamic indicators [partial pressure of oxygen (PaO 2), partial pressure of carbon dioxide (PaCO 2), oxygenation index (PaO 2/FiO 2)] before surgery and at 1 day after surgery, respiratory function [forced vital capacity (FVC), forced expiratory volume in one second (FEV1), maximal voluntary ventilation (MVV)] at 1, 3, 6 and 12 months after surgery and postoperative complications were compared between the two groups. Results:All patients were followed up for 12-18 months [(14.1±1.9)months]. Thoracoscopy group showed prolonged operation time [(139.5±36.4)minutes vs. (114.8±32.5)minutes], reduced intraoperative blood loss [(124.6±42.4)ml vs. (198.6±62.6)ml] as well as shortened ventilation time [(4.0±1.1)days vs. (6.7±1.6)days] and postoperative length of hospital stay [(14.9±2.4)days vs. (17.9±3.7)days] when compared with thoracotomy group (all P<0.01). There was no statistical significance in the number of fixed ribs between the two groups ( P>0.05). There were no statistical differences in PaO 2, PaCO 2 or PaO 2/FiO 2 between the two groups before surgery (all P>0.05). At day 1 after surgery, the PaO 2 and PaO 2/FiO 2 in thoracoscopy group were (86.2±5.4)mmHg and 321.4±36.1, higher than (80.1±6.2)mmHg and 286.0±29.3 in thoracotomy group (all P<0.01); the PaCO 2 was (37.4±2.4)mmHg in thoracoscopy group, lower than (40.0±3.1)mmHg in thoracotomy group ( P<0.01). At 1 month, 3 months, 6 months and 12 months after surgery, the FVC was (75.5±10.9)%, (84.5±10.5)%, (93.1±12.8)% and (102.6±17.5)% in thoracoscopy group, higher than (69.2±9.9)%, (78.3±8.9)%, (86.2±10.4)% and (92.4±14.8)% in thoracotomy group; the FEV1 was (76.9±9.3)%, (88.4±12.9)%, (92.4±13.9)% and (98.5±10.6)% in thoracoscopy group, higher than (72.9±8.5)%, (82.8±11.4)%, (86.4±12.7)% and (93.5±11.9)% in thoracotomy group; the MVV was (78.3±13.4)L/min, (87.5±13.5)L/min, (94.6±14.7)L/min and (100.1±11.9)L/min in thoracoscopy group, higher than (72.5±11.6)L/min, (80.5±12.7)L/min, (86.5±13.5)L/min and (92.8±10.3)L/min in thoracotomy group (all P<0.05). There were no thoracic deformities in the two groups after surgery. There was no statistical significance in incision infection rate between the two groups ( P>0.05). The incidence rate of pulmonary infection, atelectasis and pleural effusion was 11.1% (5/45), 6.7% (3/45) and 11.1% (5/45) in thoracoscopy group, lower than 29.3% (12/41), 24.4% (10/41) and 31.7% (13/41) in thoracotomy group (all P<0.05). Conclusion:Although internal fixation of ribs with complete thoracoscopy has longer surgical time than thoracotomy in the treatment of flail chest, it can decrease intraoperative blood loss, ventilation time and length of hospital stay and is more conducive to improving the respiratory function and reducing complication rate.
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Provirus integration site Moloney murine leukemia virus (Pim-1) is a proto-oncogene reported to be associated with cell proliferation, differentiation and survival. This study was to explore the neuroprotective role of Pim-1 in a rat model subjected to optic nerve crush (ONC), and discuss its related molecules in improving the intrinsic regeneration ability of retinal ganglion cells (RGCs). Immunofluorescence staining showed that AAV2- Pim-1 infected 71% RGCs and some amacrine cells in the retina. Real-time PCR and Western blotting showed that retina infection with AAV2- Pim-1 up-regulated the Pim-1 mRNA and protein expressions compared with AAV2-GFP group. Hematoxylin-Eosin (HE) staining, γ-synuclein immunohistochemistry, Cholera toxin B (CTB) tracing and TUNEL showed that RGCs transduction with AAV2-Pim-1 prior to ONC promoted the survival of damaged RGCs and decreased cell apoptosis. RITC anterograde labeling showed that Pim-1 overexpression increased axon regeneration and promoted the recovery of visual function by pupillary light reflex and flash visual evoked potential. Western blotting showed that Pim- 1 overexpression up-regulated the expression of Stat3, p-Stat3, Akt1, p-Akt1, Akt2 and p-Akt2, as well as βIII-tubulin, GAP-43 and 4E-BP1, and downregulated the expression of SOCS1 and SOCS3, Cleaved caspase 3, Bad and Bax. These results demonstrate that Pim-1 exerted a neuroprotective effect by promoting nerve regeneration and functional recovery of RGCs. In addition, it enhanced the intrinsic regeneration capacity of RGCs after ONC by activating Stat3, Akt1 and Akt2 pathways, and inhibiting the mitochondrial apoptosis pathways. These findings suggest that Pim-1 may prove to be a potential therapeutic target for the clinical treatment of optic nerve injury.
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Objective@#Preliminary study on the clinical effect of preoperative ultrasound endoscopy combined with staining labeling technique to locate the actual boundary of esophageal and gastric cancer@*Methods@#From September 1, 2015 to October 30, 2017, 18 patients with esophageal adenocarcinoma were enrolled in this study. The actual boundaries of esophageal and gastric-derived adenocarcinoma lesions were localized by endoscopic ultrasonography and staining. There were 10 males and 8 females. After completing the preoperative examination, 1-2 days before operation, endoscopic ultrasonography was used to locate the edge of the lesion. Two point injection of carbon nano suspension was used to mark the location of 1cm at the longest distance from the longitudinal axis of the tumor. According to the length of longitudinal axial staining, the thoracotomy was performed. Intraoperative proximal margin resection was used to send frozen pathology. According to the results of freezing, the operation was decided. After the operation, the specimens from the margin of the tumor were segmented into paraffin section, which was about 0.5cm in each segment, and the tumor cells were observed under the electron microscope at all levels of the paraffin sections.@*Results@#The average time of preoperative endoscopic ultrasonography staining was(10.16±1.38) min, and the diameter of nano carbon diffusion was(1.43±0.41)cm. All patients in the operation could clearly see the nano carbon staining area under the naked eye. In the field, the average time of locating lesions was(1.27±0.53)min. 5 patients underwent thoracoabdominal surgery and 13 underwent abdominal surgery. The average length of the cut margin of the tumor was(4.74±1.12)cm, and the frozen pathology of the incision margin was negative, and no additional operation was performed. The routine pathology confirmed that all the specimens were negative.@*Conclusion@#The staining and labeling technique for adenocarcinoma of the esophagogastric junction under endoscopic ultrasonography can detect the tumor edge and the scope of invasion accurately. It provides guidance and guarantee for the smooth implementation of AEG precision surgery. It is a safe, rapid and effective positioning technique.
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Objective Preliminary study on the clinical effect of preoperative ultrasound endoscopy combined with stai-ning labeling technique to locate the actual boundary of esophageal and gastric cancer Methods From September 1, 2015 to October 30, 2017, 18 patients with esophageal adenocarcinoma were enrolled in this study. The actual boundaries of esophage-al and gastric-derived adenocarcinoma lesions were localized by endoscopic ultrasonography and staining. There were 10 males and 8 females. After completing the preoperative examination, 1-2 days before operation, endoscopic ultrasonography was used to locate the edge of the lesion. Two point injection of carbon nano suspension was used to mark the location of 1cm at the lon-gest distance from the longitudinal axis of the tumor. According to the length of longitudinal axial staining, the thoracotomy was performed. Intraoperative proximal margin resection was used to send frozen pathology. According to the results of freezing, the operation was decided. After the operation, the specimens from the margin of the tumor were segmented into paraffin section, which was about 0. 5cm in each segment, and the tumor cells were observed under the electron microscope at all levels of the paraffin sections. Results The average time of preoperative endoscopic ultrasonography staining was(10. 16 ± 1. 38) min, and the diameter of nano carbon diffusion was(1.43 ±0.41)cm. All patients in the operation could clearly see the nano carbon staining area under the naked eye. In the field, the average time of locating lesions was(1.27 ±0.53)min. 5 patients under-went thoracoabdominal surgery and 13 underwent abdominal surgery. The average length of the cut margin of the tumor was(4. 74 ±1.12)cm, and the frozen pathology of the incision margin was negative, and no additional operation was performed. The routine pathology confirmed that all the specimens were negative. Conclusion The staining and labeling technique for adeno-carcinoma of the esophagogastric junction under endoscopic ultrasonography can detect the tumor edge and the scope of invasion accurately. It provides guidance and guarantee for the smooth implementation of AEG precision surgery. It is a safe, rapid and effective positioning technique.
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Objective To analyze the laboratory determination results of hand-foot-mouth disease(HFMD)and explore its sig-nificance.Methods Pharyngeal swab specimens were collected from 502 children with HFMD(severe case:104 cases and mild case:398 cases).Specific RNA of enterovirus primer(EV),enterovirus71 (EV71)and coxsackievirus A16(CA16)were detected by real-time fluorescence polymerase chain reaction(RT-PCR).Simultaneously,white blood cell (WBC),direct bilirubin(DBIL),alanine transarninase(ALT),creatine kinase MB isoenzyme(CK-MB)and lactic dehydrogenase (LDH )were determined,and the results were analyzed.Results Among the 502 children,384 cases were positive for EV,262 cases were positive for EV71,and 39 cases were positive for CA16.The positive rates were 76.49%,52.19% and 7.76%,respectively.In the 104 HFMD severe cases,EV71 positive rate was 92.31%(96/104).There were clearly more boys than girls.The age distribution was mainly in the group under 3-year-old(402 cases,80.08%).In the group under 3-year-old,there were 97 severe cases(93.27%),and the EV71 positive rate was 92.78%(90/97).Serum levels of WBC,DBIL,ALT,CK-MB and LDH were higher in severe HFMD group than in mild group(P <0.05).Conclusion The HFMD occurred mostly at the children under 3-year-old.The HFMD appeared is mainly related to the EV infection,and EV71 is the main pathogen causing the severe cases of HFMD.
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Objective To compare the efficacy of transumbilical-ultrafine gastroscope sympathectomy for severe palmar hyperhidrosis using two distinct levels of T3 and T4.Methods A total of 84 cases with severe primary hyperhidrosis were randomly allocated to undergo either T3 sympathectomy treatment (T3 group, n =42) or T4 sympathectomy treatment (T4 group, n =42) with no difference between the two groups.The operation time, postoperative hospital stay, patient's hands hyperhidrosis, axillary hyperhidrosis,complications were recorded at follow-up in 1,3,6,12 months.Results Operation on 84 patients were successful with mean operative time of T3 group being 55.02 ± 10.61 min and T4 group being 55.36 ± 10.51 min(P ＞ 0.05).The mean postoperative hospital stay were both 1.5 days.Patients were followed up for diaphragmatic hernia, umbilical hernia, Horner's syndrome and other serious complications for 12 months.No postoperative recurrence of palmar hyperhidrosis, severe compensatory sweating occurred in either group.The number of improved patients in T3 group's palmar hyperhidrosis,axillary hyperhidrosis, and foot hyperhidrosis were :42/42 cases, 10/16 cases, 21/29 cases, while those in T4 group were: 42/42 cases,16/17 cases 18/28 cases.Axillary hyperhidrosis improved to a larger extent in T4 group than in T3 group (P ＜0.05).There were 16(15 mild and 1 moderate)compensatory sweating in T3 group and 7(6 mild and 1 moderate) compensatory sweating in T4 group (P ＜ 0.05).Conclusion T3 and T4 thoracic sympathectomy using transumbilical-ultrafine gastroscope for primary palmar hyperhidrosis are safe, effective, and feasible.T4 sympathectomy is more effective in improving axillary hyperhidrosis than T3 and shows lower occurrence of postoperative compensatory hyperhidrosis.
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Objective To evaluate the safety and feasibility of endoscopic transumbilical thoracic sympathectomy.Methods The technique was performed in 4 swine models.Under general anesthesia,a newly developed long abdominal trocar (60 cm in lengths) was placed through the umbilicus,which was also used to establish pneumoperitoneum.After insertion of gastroscope through this trocar,two small incisions of diagrams were created by needle-knife,through which the gastroscope was sent into the thoracic cavity to ablate the T3 ganglia by Hot biopsy forceps under One-lung ventilation.The animals were killed at the completion of the procedure.Results This procedure was completed in four pigs.The sympathetic chain was successfully ablated in all swine,as confirmed by gross surgical pathology and histology,the mean operative time was (81.7 ±15.4) minutes.There was no major bleeding.All bilateral T3 sympathetic chains were successfully ablated in the 4 swines as confirmed by gross surgical pathology and histology.There was no severe bleeding and damage to peripheral organs during the operation in any of the cases,and no prominent pneumothorax was revealed by postoperative chest X-ray radiography.Conclusion Endoscopic transumbilical thoracic sympathectomy is safe,feasible and effective with good cosmetic result.
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Objective To evaluate the technique of finger palpation in thoracoscopic localization in patients with pulmonary nodules,and to summarize its technical details,especially with exploit of chest computed tomography (CT) facilitating it.Methods 95 patients with total amount of 109 pulmonary nodes 20 mm or smaller in size shown with lung window of CT,were reviewed.They were located subpleurally,with a median depth of 8.2 mm and a median size of 10.0 mm.The value of their depth over their size (D/d value) could be used as the extent of localizing difficulty.Each node had its own radiographic fealures for being localized,which was built preoperatively.Under thoracoscopic vision,nodules were finger-palpated by index finger via the 4th or 5th intercostal space on anterior axillary line,followed by wedgectomy or lobectomy for instant histopathological diagnosis to further decide the final surgical type.The distance between the nodule and the origin of segmental bronchus (L value) were also calculated out,as it might be relevant to the way the nodule could be biopsied.Results All nodules were successfully localized and resected for biopsy goal,105 by wedgectomy,4 by lobectomy.After intraoperative diagnosis was made by the pathologist,VATS lobectomy and lymph node dissection were further performed in 55 patients.L value of 4 cases being biopsied by lobectomy ranged from 18.3 to 30.3 mm,averaging 26.1 mm.Conclusion Finger palpation is viable in any cases of pulmonary nodules.Detailed reference of CT digital information,and enough detachment of mediastinal pleura,can greatly facilitate thoracoscopic localization by finger palpation.Lobectomy or segementectomy is preferable when L value is less than 30 mm.
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To study the effects of puerarin on the aromatase P450 (P450(arom)) mRNA expression and the effects of low-dose puerarin on transcription factors of the P450(arom) gene (P II) 5'-flanking region.
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Objective: To clone hGLP-1 cDNA in the pBS SK(+/-)vector and construct the expression vector of pGEX-4T-3/hGLP-1cDNA to express GST-hGLP-1 fusion protein. Methods: The hGLP-1 cDNA was constructed by 6 synthetic oligonucleotides fragments, followed by the procedure of annealing and ligation with oligonucleotides fragments. The hGLP-1 cDNA was cloned into the pBS SK(+/-) vector, and was selected by α-complementation. It was confirmed by DNA sequening, then inserted into the MCS of the fusion expression vector pGEX-4T-3. The recombinant vector was transformed into E. coli TG1. Results: The recombinant plasmid DNA was digested with restrictive endonuclease BamHⅠand XhoⅠ. The result demonstrated that the hGLP-1 cDNA was successfully inserted into the pGEX-4T-3 vector and fusion protein GST-hGLP-1 had been expressed in SDS-PAGE. Conclusion： Expression of GST-hGLP-1 fusion protein can provide foundation for obtaining a larger quantity of recombinant hGLP-1 for experimental and clinic studies.
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Objective:To construct eukaryotic inducible expression plasmids pTRE-Tight-CFP-CD11b and pTRE-Tight-YFP-CD18 using Tet-On gene expression system and co-transfect them into Chinese hamster ovary(CHO) cells,so as to achieve the inducible expression of Mac-1-FP.Methods: The eukaryotic expression vectors pTRE-Tight-CFP-CN11b and pTRE-Tight-YFP-CD18 were constructed by recombinant DNA technique.The 2 vectors were co-transfected into CHO cells using liposome to fuse CD11b and CD18: the 2 subunits of Mac-1. Fluorescence microscope was used to observe the cyan fluorescence and the yellow fluorescence of Mac-1-FP.The influence of different levels of Dox(0,0.01, 0.1,0.5,1,2 ?g/ml) on expression levels of CD11b and CD18 in CHO cells was analyzed by RT-PCR and fluorescence intensity analysis.The adhesive rate of CHO-Mac-1-FP cells with ligand ICAM-1 was analyzed before and after PMA(1 ?g/ml) stimulation.Results: The recombinant plasmids of pTRE-Tight-CFP-CD11b and pTRE-Tight-YFP-CD18 were successfully constructed.The cyan and yellow fluorescences were observed in co-transfected CHO cells under fluorescence microscope.The fluorescence intensity of the cells was increased with the increase of Dox concentration.RT-PCR analysis demonstrated that the CD11b and CD18 mRNA increased with the increase of Dox concentration.The adhesive rate of CHO-Mac-1-FP cells with ICAM-1 was increased after PMA stimulation(peaked at 2 h and 4 h after stimulation and decreased afterwards).Conclusion: This study achieves the inducible expression of Mac-1-FP in CHO cells.And Mac-1-FP,like widetype Mac-1,exhibits adhesive activity with ligand ICAM-1,which lays a foundation for studying the consisting dimmer,clustering,conformation and affinity of the ligands of Mac-1 using single molecule spectroscopy and fluorescence resonance energy transfer technique in living cells.