1.In vitro experiment of stem cell engineered two-sided anisotropic electrospun membranes for promoting dural repair
Jingzhi XU ; Wenbo WANG ; Huiwen SUN ; Yong GU
Chinese Journal of Tissue Engineering Research 2024;28(10):1540-1546
BACKGROUND:Currently,the dura mater is clinically repaired using autologous tissue or materials such as gelatin sponge,but all of them have their inherent defects.Therefore,there is an urgent need for a biomaterial that can promote dural repair. OBJECTIVE:The two-sided anisotropic electrospun membrane was constructed by using directional electrospinning technology and collagen self-assembly technology,and was used as a carrier for bone marrow mesenchymal stem cells to investigate various physicochemical properties and biological characteristics of the artificial dura mater. METHODS:Ordered polylactic acid electrospun fibers with double-sided(collagen protein on one side and polylactic acid on the other side)anisotropic electrospun membranes(collagen group),disordered polylactic acid electrospun membranes(disordered fiber group),and ordered oriented polylactic acid electrospun membranes(ordered fiber group)were prepared by electrospinning technique as well as collagen self-assembly technique.Scanning electron microscopy,mechanical stretching,water contact angle testing,and degradation experiments were used to characterize the physicochemical properties of the electrospun membranes.Electrospun membranes in the collagen group(bone marrow mesenchymal stem cells were inoculated on the collagen surface to obtain the stem cell-engineered electrospun membranes),disordered fiber group and ordered fiber group were cocultured with bone marrow mesenchymal stem cells.The biocompatibility of electrospun membranes was evaluated using CCK-8 assay and live/dead staining.Integrin β1 immunofluorescence staining was used to evaluate the adhesion characteristics of electrospun membranes.The stem cell-engineered electrospun membrane and the electrospun membrane in the collagen group were cocultured with bone marrow macrophages respectively.Immunomodulatory properties were assessed by detecting the expression of inflammation-related genes using inducible nitric oxide synthase(M1 type),CD206(M2 type)immunofluorescence staining,and qRT-PCR. RESULTS AND CONCLUSION:(1)The oriented electrospun fiber membrane could mimic the structure of the longitudinally aligned natural dura mater,and the addition of collagen increased the hydrophilicity of the fiber membrane by about 2-fold and the mechanical properties by 1.2-fold.(2)When cocultured with bone marrow mesenchymal stem cells,CCK-8 assay and live/dead staining suggested that the cellular bioactivity in the collagen group was significantly higher than that in the disordered fiber group and ordered fiber group.Immunofluorescence staining revealed that the expression of integrin β1 in the collagen group was about 2.6 times higher than that of the disordered and ordered fiber groups,and the cell spreading morphology was good.(3)When cocultured with bone marrow macrophages,immunofluorescence staining exhibited that the fluorescence intensity of M1 type macrophages in the stem cell-engineered electrospun membrane group was lower than that in the collagen group(P<0.01),and the fluorescence intensity of M2 type macrophages was higher than that in the collagen group(P<0.01).qRT-PCR demonstrated that proinflammatory gene tumor necrosis factor α and interleukin-1β mRNA expression in the stem cell-engineered electrospun membrane group was lower than that of the collagen group(P<0.001);anti-inflammatory genes such as interleukin-10 and transforming growth factor β mRNA expressions were higher than those in the collagen group(P<0.001).(4)The above results suggest that the stem cell-engineered amphipathic artificial dura mimics the directional structure of normal dura,with the inner surface facilitating cell growth and adhesion and the outer edge avoiding tissue adhesion,while the polarization of macrophages to the M2 subtype is promoted and the local inflammatory microenvironment is regulated through the mesenchymal stem cell paracrine component.
2.Role of AQP4 in dexmedetomidine-induced reduction of blood-brain barrier permeability in mechanically ventilated mice: relationship with PKC
Min QU ; Wenbo SUN ; Xiuqing ZHANG ; Wang LIU ; Lei CHEN ; Zilong QI ; Dongdong HUANG
Chinese Journal of Anesthesiology 2024;44(3):318-323
Objective:To evaluate the role of aquaporin 4 (AQP4) in dexmedetomidine-induced reduction of blood-brain barrier permeability in mechanically ventilated mice and the relationship with protein kinase C (PKC).Methods:One hundred and fifty clean-grade healthy male C57BL6 mice, weighing 20-25 g, aged 8-12 weeks, were divided into 5 groups ( n=30 each) using a random number table method: control group (group C), mechanical ventilation group (group V), LY317615 group (group L), dexmedetomidine group (group D), and dexmedetomidine+ PMA group (group DP). Group C spontaneously breathed air for 6 h. The animals were mechanically ventilated for 6 h in group V. PKC inhibitor LY3176 15 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation in group L. Dexmedetomidine 50 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation in D and DP groups. PKC activator PMA 15 μg/kg was intraperitoneally injected at 60 min before mechanical ventilation in group DP. Mice were anesthetized at 1 day after mechanical ventilation, then sacrificed and hippocampal tissues were taken for microscopic examination of pathological changes in the hippocampal CA1 and CA3 areas (with a light microscope). Brain tissues were also taken to measure the water content and content of Evans blue (EB) and to detect the expression of PKC and AQP4 (by Western blot). The cognitive function was evaluated using a novel object recognition task at 3 days after mechanical ventilation. Results:Compared with group C, the water content and EB content of brain tissues were significantly increased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was up-regulated, the percentage of novel object exploration and discrimination index were decreased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was aggravated in group V and group DP. Compared with group V, the water content and EB content of brain tissues were significantly decreased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was down-regulated, the percentage of novel object exploration and discrimination index were increased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was significantly attenuated in group D and group L. Compared with group D, the water content and EB content of brain tissues were significantly increased after mechanical ventilation, the expression of PKC and AQP4 in brain tissues was up-regulated, the percentage of novel object exploration and discrimination index were decreased ( P<0.05), and the histopathological damage in the hippocampal CA1 and CA3 areas was aggravated in group DP. Conclusions:AQP4 is involved in dexmedetomidine-induced reduction of blood-brain barrier permeability in mechanically ventilated mice, and the mechanism is related to inhibiting activation of PKC.
3.Study on Acute Toxicology and Intervention Effect of Panacis Majoris Rhizoma on Chronic Pharyngitis Rat Model
Xudong HE ; Lianli ZHOU ; Lankun YANG ; Yuxuan TAO ; Zhibo LIU ; Wenbo WANG ; Siqi SUN ; Jie YU ; Xingxin YANG
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(1):10-16
Objective To investigate the acute toxicology and intervention effect of Panacis Majoris Rhizoma on rats with chronic pharyngitis.Methods A single,maximum dose of Panacis Majoris Rhizoma(74.4 g·kg-1)was administered to Kunming mice to evaluate its toxicity,involving the assessment of the survival status of the mice,organ indices,morphological changes in major organs,blood routine,and biochemical indicators.SD rats were randomly divided into the control group,model group,prednisone group(6.25 mg·kg-1),and low-,medium-,and high-dose Panacis Majoris Rhizoma groups(0.58,1.16,and 2.32 g·kg-1).All rats received the corresponding drugs(or normal saline)via intragastric administration once daily for a duration of 30 days.Except the control group,chronic pharyngitis was induced in rats of the other groups by using β-hemolytic streptococcus.Following euthanasia,serum inflammatory levels of interleukin-6(IL-6),cyclooxygenase-2(COX-2),interleukin-1β(IL-1β),intercellular adhesion molecule-1(ICAM-1),C-reactive protein(CRP),tumor necrosis factor(TNF-α),monocyte chemoattractant protein-1(MCP-1),and prostaglandin E2(PGE2)were measured.Additionally,pharyngeal tissues were stained with HE and pathological characteristics were observed.Results Toxicological studies have demonstrated that the administration of Panacis Majoris Rhizoma resulted in significant increase in plasma alanine transaminase levels and spleen index of mice,along with corresponding tissue pathological alterations.Nevertheless,no noteworthy pathological changes were observed in other organs,and there were no notable changes in blood routine and plasma biochemical indicators.Pharmacodynamic investigations have revealed that Panacis Maioris Rhizoma effectively reduces the serum levels of inflammatory factors and improves pathological changes in pharyngeal tissues.Conclusion Panacis Maioris Rhizoma alleviated β-hemolytic streptococcus-induced CP by inhibiting inflammatory responses,and may show potential toxicity to the spleen.
4.Treating Emotional Disorders from the Classification of Gallbladder
Yijun WU ; Yongkang SUN ; Fangbiao XU ; Yanbo SONG ; Wenbo LIU ; Shuang JIANG ; Haohao LIAO ; Xinzhi WANG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(3):309-314
This article summarizes the unique viewpoints and experience application of the famous and veteran Chinese medicine practitioner,Professor Wang Xinzhi,in treating emotional diseases from the perspective of gallbladder theory.Based on the physiologi-cal functions and characteristics of the gallbladder in Chinese medicine,it is proposed that the"heart mind-gallbladder-viscera"axis dominates the generation and changes of emotions,and it is believed that gallbladder failure is the key pathogenesis of emotional disor-ders.The treatment of clinical syndromes should be based on the type of gallbladder,and emotional diseases can be divided into types of insufficient gallbladder qi,unfavorable Shaoyang,gallbladder and heat excess,timidity-deficiency,and heart-gallbladder indeci-sion,according to clinical manifestations;based on the basic principle of adjusting the functions of the heart,spleen,liver,gallblad-der,kidney and other organs,treatment methods such as tonifying the spleen and kidneys,increasing gallbladder qi,resolving Shaoy-ang,clearing gallbladder heat,warming yang and replenishing qi,calming the mind,resolving phlegm and removing blood stasis should be used,highlighting the joint treatment of the heart and the gallbladder,and the simultaneous regulation of the liver and gall-bladder,so that the mind can be at ease,the gallbladder can be decisive,and the emotions can be harmonious.
5.Comparison of the efficacy of neoadjuvant chemotherapy versus neoadjuvant chemoradiotherapy in locally advanced mid-low rectal cancer
Qi SUN ; Chang LIU ; Jianqiao YE ; Wenbo HUANG ; Yuhang XU ; Chengzhi YAO ; Wei ZHAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2024;45(2):278-283
【Objective】 To investigate the differences in efficacy and long-term prognosis between locally progressive low and intermediate rectal cancer patients receiving fluorouracil-based neoadjuvant chemotherapy alone (mFOLFOX6/CapeOX) and neoadjuvant radiotherapy, and to compare the therapeutic efficacy in the two groups. 【Methods】 We retrospectively analyzed the clinicopathological data of 118 patients with locally progressive low and intermediate rectal cancer who received neoadjuvant therapy from January 2019 to December 2021 at The First Affiliated Hospital of Xi’an Jiaotong University, including gender, age, body mass index (BMI), and other clinicopathological parameters. The t-test, Mann Whitney test, chi-square test or Fisher’s exact test were used to compare the differences between the two groups of patients who received neoadjuvant chemotherapy alone or neoadjuvant radiochemotherapy in terms of short-term efficacy, lymph node manifestations and long-term prognosis, respectively. Survival rates were calculated and survival curves were plotted using the Kaplan-Meier method. 【Results】 In terms of efficacy, patients in the neoadjuvant radiotherapy group achieved better tumor regression (Z=-2.05, P=0.04) and solid tumor efficacy (Z=-2.42, P=0.015), but the difference between the two groups in terms of downstaging effect of clinical stage was not statistically significant. The number of lymph nodes detected was significantly lower in the neoadjuvant radiotherapy group (neoadjuvant chemotherapy vs. neoadjuvant radiochemotherapy, 13.19±3.83 vs. 9.55±4.00, t=5.02, P<0.001), but the two groups did not differ significantly in the number of lymph node positives and lymph node positive ratio. In terms of long-term prognosis, there was no statistically significant difference in the overall survival rate or disease-free survival rate of the two groups. 【Conclusion】 Compared with neoadjuvant chemotherapy alone, neoadjuvant radiotherapy showed better short-term efficacy in patients with locally progressive low and intermediate rectal cancer, but there was no statistically significant difference between the two treatment regimens in terms of long-term prognosis.
6.Understanding the excellent use of Chaihu Guizhi Decoction from xiang thinking
Yilong SUN ; Fangbiao XU ; Yanbo SONG ; Yuhe HU ; Yongkang SUN ; Wenbo LIU ; Liuping YUE ; Xinzhi WANG
Journal of Beijing University of Traditional Chinese Medicine 2024;47(11):1512-1519
Chaihu Guizhi Decoction is an excellent prescription of ZHANG Zhongjing;however,nowadays the application of Chaihu Guizhi Decoction is primarily derived from the text.The xiang thinking is the source of traditional Chinese medicine thinking.Understanding Chaihu Guizhi Decoction from the xiang thinking,it is easy to perceive the true meaning of ZHANG Zhongjing,and grasp the location of the disease and the patient's overall qi,so as to make better use of this formula.This article discusses the understanding of Chaihu Guizhi Decoction from five perspectives:firstly,it gives an overview of the xiang thinking,pointing out the source and essential value of the xiang thinking;secondly,it analyses the disease position and the patient's qi of Chaihu Guizhi Decoction under the perspective of the xiang thinking as a whole;thirdly,it discusses the disease position and the patient's qi of the original text of Chaihu Guizhi Decoction from the xiang thinking;fourthly,it discusses how to understand the principle of formula formation and the main and minor parts of Chaihu Guizhi Decoction by xiang thinking;finally,it analyses the primary medicines in Chaihu Guizhi Decoction by using xiang thinking based on the records of Shennong Bencao Jing.Based on these five points,we hope to reveal the traditional Chinese medicine thinking behind Chaihu Guizhi Decoction,and provide new ideas for the clinical application of Chaihu Guizhi Decoction.
7.Correlation between the Expression of LNMAS mRNA and CLDN4 mRNA Levels in Cervical Cancer Tissue and Epithelial-Mesenchymal Transition and Clinical Significance
Chunwen LIU ; Yue WANG ; Xinyue LIU ; Wenbo XU ; Zhimin SUN
Journal of Modern Laboratory Medicine 2024;39(6):90-95
Objective To investigate the correlation between the expression of long non-coding RNA lymph node metastasis-associated suppressor(LNMAS)mRNA and tight junction protein claudin 4(CLDN4)mRNA and epithelial-mesenchymal transition(EMT)in cervical cancer and the clinical significance.Methods A total of 96 patients with cervical cancer who were first diagnosed and treated at Cangzhou Maternal and Child Health Hospital from June 2018 to June 2020 were selected.Real-time fluorescent quantitative PCR was used to detect the expressions of LNMAS mRNA,CLDN4 mRNA and EMT-related genes.Pearson correlation analysis was used to analyze the correlation between LNMAS mRNA,CLDN4 mRNA and VIM mRNA,E-cad mRNA,N-cad mRNA.The impact of LNMAS mRNA and CLDN4 mRNA expression on the 3-year survival rate of cervical cancer patients was analyzed using the K-M curve.Cox regression was used to analyze prognostic factors for cervical cancer.Results Compared to adjacent tissues,the expressions of LNMAS mRNA(1.13±0.28 vs.1.97±0.37)and E-cadherin(E-cad)mRNA(1.02±0.33 vs 2.29±0.56)in cervical cancer tissues were lower(t=17.738,19.144),while CLDN4 mRNA(3.14±0.52 vs 1.19±0.28),N-cadherin(N-cad)mRNA(2.60±0.36 vs 1.02±0.33)and Vimentin(VIM)mRNA(2.84±0.46 vs 1.25±0.33)were higher(t=32.351,26.951,27.518),with significant differences(all P<0.05).LNMAS mRNA showed a positive correlation with E-cad mRNA(r=0.712,P<0.05),but a negative correlation with VIM mRNA and N-cad mRNA in cervical cancer tissues(r=-0.654,-0.589,all P<0.05).Additionally,the expression of CLDN4 mRNA was negatively correlated with E-cad mRNA(r=-0.668,P<0.05),but positively correlated with VIM mRNA and N-cad mRNA(r=0.714,0.749,all P<0.05).Compared with patients with FIGO stage ⅠA~ⅠB1 and no lymph node metastasis,patients with FIGO stage ⅠB2-ⅡA and lymph node metastasis exhibited a lower LNMAS mRNA expression and a higher CLDN4 mRNA expression with significant differences(t=12.288,10.228,13.636,11.771).Comparison Log-Rank x2 square test,the 3-year overall survival rates for high and low expression groups of LNMAS mRNA were 92.00%(46/50)and 60.87%(28/46),respectively;the 3-year overall survival rates for high and low expression groups of CLDN4 mRNA were 59.18%(29/49)and 95.74%(45/47),respectively,and the difference between the two groups were significant(log-Rank x2=11.030,15.600,all P<0.001).FIGO stage ⅠB2~ⅡA(HR=1.119,95%CI:1.148~1.830),lymph node metastasis(HR=1.442,95%CI:1.124~1.850)and CLDN4 mRNA(HR=1.637,95%CI:1.124~1.850)were risk factors affecting the prognosis of cervical cancer patients,while LNMAS mRNA(HR=0.637,95%CI:0.465~0.873)was a protective factor(all P<0.05).Conclusion The expression of LNMAS mRNA is decreased,while the expression of CLDN4 mRNA is increased in cervical cancer.The expressions of LNMAS mRNA and CLDN4 mRNA were associated with the process of EMT,which may be markers for evaluating the prognosis of patients with cervical cancer.
8.Research on patient motion monitoring with domestic innovative integrated radiotherapy CybeRay ? real-time imaging for frameless stereotactic radiosurgery
Lihong CAI ; Wenbo GUO ; Jing NIE ; Yali WU ; Minjie ZHANG ; Huina SUN ; Xinsheng XU ; Gaoqing FENG ; Rui ZHANG ; Qingfang JIANG ; Yu ZHANG ; Yubing XIA
Chinese Journal of Radiation Oncology 2024;33(12):1138-1143
Objective:To determine the motion detection uncertainty of the real-time CybeRay ? imaging system and patient intrafractional motion with thermoplastic mask-based immobilization. Methods:Real-time CybeRay ? imaging system was used for irradiation and treatment for head phantom and patients with brain tumors. All patients were immobilized with thermoplastic masks. Real-time imaging was delivered using kilovoltage projection images during radiotherapy. The detected patient motion data was collected from 5 head phantom measurements and 27 treatment fractions of 9 brain tumor patients admitted to Kaifeng Cancer Hospital. The accuracy and uncertainty of the motion monitoring system were determined. Results:The mean and standard deviation (SD) of the detected motion in the X, Y, and Z directions for phantom were (-0.02±0.41) mm, (-0.05±0.22) mm and (0.01±0.35) mm, respectively. The detected motion in the X, Y and Z directions for patents were (-0.13±0.48) mm, (-0.05±0.48) mm and (0.11±0.36) mm, respectively. After removing the motion detection uncertainty, the actual intrafractional motion of patients were (-0.11±0.25) mm, (0±0.43) mm and (0.10±0.08) mm in three directions, respectively. Conclusions:The uncertainty of real-time imaging-based motion monitoring system of CybeRay ? is less than 0.5 mm. It is feasible to apply thermoplastic masks for brain tumor patients in clinical practice, which can provide steady immobilization and limit the SD of patient intrafractional motion within 0.5 mm. Real-time imaging-based motion monitoring system of CybeRay ? is accurate for patient motion monitoring during frameless stereotactic radiosurgery/radiotherapy.
9.Preliminary Experience of Needle Grasper Hydrodissection Assisted Laparoscopic Orchiopexy
Xiaofeng YANG ; Chi SUN ; Wenbo WANG ; Hui ZHOU ; Meng LI ; Yazhen MA ; Suolin LI
Chinese Journal of Minimally Invasive Surgery 2024;24(6):451-454
Objective To investigate the safety and effectiveness of laparoscopic orchiopexy assisted by needle grasper hydrodissection for cryptorchidism.Methods From September 2020 to September 2022,21 children with cryptorchidism on 25 sides underwent laparoscopic orchiopexy assisted by needle grasper hydrodissection in our department.The water injection function of needle grasper was applied to inject normal saline into the retroperitoneal space to fully separate the retroperitoneum from the vas deferens and spermatic cord,establishing a liquid barrier to protect the vas deferens and spermatic cord.Then the retroperitoneum was bridge-likely cut off with the assistance of needle grasper.The vas deferens and spermatic cord were completely released in accordance with the principle of integrity and non-destructiveness,then the testes were successfully induced and fixed.Results During the operation,15 cases of inguinal cryptorchidism and 6 cases of intra-abdominal cryptorchidism were confirmed.There were 17 cases of unilateral cryptorchidism and 4 cases of bilateral cryptorchidism.All the 25 testes were successfully re-located into the dartos pouch of the scrotum under laparoscopic guidance.The operation time was 35-75 min(mean,45.1±14.3 min),and there were no surgical complications.During follow-ups for 12-24 months in 21 cases,the testicles were found located in the scrotum without retraction,atrophy,incision infection,incision hernia,or inguinal hernia.Conclusions Needle grasper hydrodissection assisted laparoscopic orchiopexy can effectively protect the vas deferens and spermatic cord.The operation is simple,safe,and effective.
10.Surgical techniques for pollybeak deformity correction
Hongli ZHAO ; Xiancheng WANG ; Yang SUN ; Xiang XIONG ; Xianxi MENG ; Wenbo LI ; Zhihua QIAO ; Kai YANG ; Weiliang ZENG ; Yi TIAN
Chinese Journal of Plastic Surgery 2024;40(2):156-162
Objective:To evaluate the effectiveness of surgical methods for correcting pollybeak deformity in Chinese rhinoplasty.Methods:A retrospective chart review was conducted for patients who underwent pollybeak correction between January 2021 and December 2022 at the Department of Plastic and Aesthetic (Burn) Surgery, the Second Xiangya Hospital of Central South University. Individualized correction was tailored based on the etiology and severity of the nasal deformity of each patient, involving techniques such as resection of the anterior part of the nasal septum, reconstruction of nasal tip support, reconstruction of the middle part of the nasal vault, and excision of skin in the upper region of the nasal tip. A modified classification system for pollybeak deformity, the supratip fullness rating scale (SFRS), was developed to evaluate supratip fullness (0-3 points, with higher scores indicating more apparent deformity). The patients aesthetic outcomes were assessed by surgeons using the visual analogue scale (VAS) (0-10 points, with higher scores indicating more apparent deformity), and patient self-assessed using the rhinoplasty outcome evaluation (ROE) questionnaire (0-100 points, with higher scores indicating higher satisfaction). The measurement data of normal distribution was expressed as Mean±SD and analyzed by paired t-test; the measurement data of non-normal distribution was expressed as M( Q1, Q3) and analyzed by Wilcoxon signed rank test. Results:In a cohort study of 53 rhinoplasty patients (7 male, 46 female; age range 19-45 years, mean 29 years), comprising 15 primary and 38 secondary surgeries, nasal tip deformities were evaluated. Deformities were classified as mild (5 cases), moderate (25 cases), and severe (23 cases). Over a follow-up period of 6-17 months (mean 9.5 months), significant aesthetic improvements in the nasal tip region were observed. The SFRS scores decreased from 2(2, 3) preoperatively to 0(0, 0) postoperatively ( Z = -6.58, P < 0.001), and VAS scores decreased from 7.47±1.73 to 1.79±1.67 ( t = -25.61, P < 0.001). High patient satisfaction was indicated by a mean ROE score of 82.45±11.55. No significant complications, such as nasal tip ptosis, skin necrosis, or scar hyperplasia, were reported. Conclusion:Selecting an appropriate surgical method based on the severity and cause of pollybeak deformity can achieve satisfactory outcomes. Post-operative patients exhibit significant aesthetic improvement in the upper nasal tip area, resulting in high patient satisfaction.

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