1.Effect of tension on collagen arrangement
Yingkai LIU ; Xiqiao WANG ; Jun WEI ; Chun QING ; Shuliang LU
Chinese Journal of Trauma 2009;25(9):779-782
Objective To study the effect of mechanical tension on collagen arrangement and illustrate the relationship between tissue architecture and tension properties. Methods Cell morphologies, orientation and collagen arrangement of fibroblasts cultured in three different types of collagen gels with variation of mechanical tension were observed by phase contrast photomicrographs, light microscopy and transmission electron microscopy. Expression and distribution of a-smooth muscle actin (α-SMA) were investigated by immunohistochemistry. Results Phase contrast photomicrographs, light microscopy and transmission electron microscopy showed high level of tension distributed anisotropically in the monolayer gels and the anchored collagen gels, with bipolar shape of the fibroblasts, obvious polarity, arrangement of exogenous collagen fibres parallel to the long axis of the fibroblasts, especially prominent in monolayer gels. Low level of tension distributed isotropically was observed in floating collagen gels, with stellate morphology and arrangement of exogenous collagen fibres in a reticular array. Immunofluorescence showed that fibroblasts expressed high level of α-SMA protein distributed along the long axis of fibroblasts in the monolayer gels and the anchored collagen gels, especially in former ones. In contrast, few expression of α-SMA protein was found in floating collagen gels. Cell morphologies and orientation, expression and distribution of α-SMA as well as collagen arrangement of fibroblasts in the monolayer gels and the anchored collagen gels were similar to those in granulation tissue, whereas floating collagen gels resembled normal dermis or remodelled tissues. Conclusions Tissue architecture or morphology of the dermis are corresponding to tension proporties. Different tissue architectures are closely correlated with particular tension proporties.
2.Histology of collagen nodules from hypertrophic scars
Yingkai LIU ; Xiqiao WANG ; Fei SONG ; Zhiyong WANG ; Bo YUAN ; Chun QING ; Shuliang LU
Chinese Journal of Trauma 2014;30(4):302-306
Objective To detect the histological characteristics of collagen nodules from hypertrophic scars (HS) and investigate the origin of collagen nodules.Methods The scar tissues were collected from patients with plastic operation.Morphological characteristics of collagen nodules were observed by light microscopy of HE-stained sections; expressions of type Ⅰ /Ⅲ collagens were observed by polarized light microscopy of sirius red-stained sections; expression and distribution of myofibroblasts (MFb)-specific protein (α-smooth muscle actin,α-SMA) were observed by immunostaining in order to observe level of local tissue tension.Results Collagen nodules varied in shape,not only sphereshaped,and in size.Moreover,abundant fibroblasts (Fb) with large and light-stained nuclei were seen in the nodules compared to non-nodule area,indicating that the cells located in the modules were active.Some collagen nodules were composed largely of collagen type Ⅲ (green),but some mainly contained collagen type Ⅰ (red or yellow),indicating the difference in the time of nodule formation.α-SMA was expressed mainly in the deep dermis equivalent to the level of reticular layer of the new scar tissues (2 months after injury) ; α-SMA was expressed mainly in the nodules of the old scar tissues (2-10 years after injury),but almost not in non-nodule areas except for a strongly positive staining in the vessels.Moreover,α-SMA presented a heterogeneous distribution in the collagen nodules,with stronger expression in the epidermal end than in the subcutaneous tissue end and stronger expression in the superficial dermis than that in the deeper part.It was suggested that there existed massive amount of MFb and high tension in the nodules arid that the tension distribution was not uniform in or between the nodules.Conclusions Collagen nodules are of varying shape and size and collagen types are associated with the time of nodule formation.Moreover,Formation of the collagen nodules may be closely related to the distribution and evolution of the local tissue tension.
3.Application of different doses of ropivacaine combined with sufentanil in epidural stepwise labor analgesia LI
Xujun CHEN ; Yan GUO ; Zhanqiang ZHAO ; Yun ZHU ; Jiaqin LI ; Xiqiao WANG
The Journal of Clinical Anesthesiology 2016;32(4):361-365
Objective To evaluate the efficacy of epidural anesthesia combined with different doses ropivacaine and sufentanil for stepwise labor analgesia in latent phase.Methods Two hundred and ten ASA Ⅰ or Ⅱ primiparas with a singleton and vertex presentation at full term in our hospital from February 201 5 to April 201 5 were randomized into seven groups (n =30 each):0.125% ropiva-caine with 0.5 μg/ml sufentanil (group 1);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm) (group 2);0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group3);0.1 5% ropivacaine with 0.5μg/ml sufentanil (group 4);0.075% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥ 3 cm)(group 5 );0.1%ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm)(group 6);0.125% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation<3 cm),0.1 5% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation≥3 cm) (group 7).The intensity of pain was assessed by visual analog scale (VAS).Meanwhile,1abor process,postpartum hemorrhage,Bromage score,postpartum adverse reactions and Apgar score of the neonates were also observed.Results No significant difference was found in VAS score after epi-
dural block between groups at each time.The latent period of group 2 and 3 were shorter than that in group 1 (P <0.05)and that of group 5 and 6 were shorter than that in group 4 (P <0.05);the ac-tive phase of group 4 were longer than that in group 1 (P <0.05 ).The postpartum hemorrhage of group 2 and 3 were less than that in group 1 (P <0.05),the postpartum hemorrhage of group 5,6 and 7 were more than that in group 2 (P <0.05)and group 3 (P <0.05).The motor nerve block of group 2 and 3 were slightly less than that in group 1 (P <0.05)and the motor nerve block of group 5,6 and 7 were slightly less than that in group 4 (P <0.05).There was no difference of the postpar-tum adverse reactions of maternal and Apgar score in the neonates.Conclusion The dosage of 0.075% or 0.1% ropivacaine with 0.5 μg/ml sufentanil (cervical dilatation < 3 cm),0.125% ropiv-acaine with 0.5 μg/ml sufentanil (cervical dilatation ≥ 3 cm),while producing the exact analgesic effect,hardly interferes with the 1abor process,the amount of postpartum hemorrhage and the lower limb activity,thus they have no significant effect on the safety of the maternal and the infant.
4. Effects of denatured collagen type Ⅰ on differentiation of human fibroblasts into myofibroblasts
Zhiyong WANG ; Xiqiao WANG ; Yingkai LIU ; Bo YUAN ; Jiaoyun DONG ; Fei SONG ; Yuzhi JIANG ; Shuliang LU
Chinese Journal of Burns 2018;34(2):96-101
Objective:
To investigate the effects of denatured collagen type Ⅰ on differentiation of human fibroblasts into myofibroblasts.
Methods:
A small amount of normal skin donated by burn patients undergoing scar surgery was collected. Human fibroblasts were obtained by method of explant culture and then sub-cultured. The fourth passage of cells were used in the following experiments. (1) Fibroblasts were divided into normal collagen group and denatured collagen group according to the random number table, with 10 wells in each group. Fibroblasts in normal collagen group were cultured on normal collagen type Ⅰ coated coverslips. Fibroblasts in denatured collagen group were cultured on denatured type Ⅰ collagen coated coverslips. Expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical method, and the percentage of PCNA positive cells was calculated. (2) Another batch of fibroblasts were grouped and treated as in (1), with 12 wells in each group. Proliferation activity of cells was determined with methyl-thiazolyl-tetrazolium colorimetry method. (3) Another batch of fibroblasts were grouped and treated as in (1), and the microfilament morphology of cells was observed by rhodamine-phalloidin staining. (4) Another batch of fibroblasts were grouped and treated as in (1). Expression of α smooth muscle actin (α-SMA) of cells was detected by immunohistochemical method, and expression of OB-cadherin of cells was detected by immunofluorescence method. (5) Another batch of fibroblasts were divided into normal collagen, denatured collagen, and common coverslips groups according to the random number table, with 6 wells in each group. Fibroblasts in normal collagen and denatured collagen groups were treated as in (1), while fibroblasts in common coverslips group were cultured on coverslips without collagen coating. Expressions of α-SMA and OB-cadherin of cells were determined with Western blotting. (6) Another batch of fibroblasts were grouped and treated as in (5), and then the mRNA expressions of collagen type Ⅰ, collagen type Ⅲ, and α-SMA of cells were determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with
5. Clinical Study on Changes of Gastric Myoelectrical Activity and Autonomic Nerve Function in Cirrhotic Patients With Gastroesophageal Varices After Endoscopic Treatment
Mengyu LI ; Yi CHEN ; Meihong LIU ; Meifeng WANG ; Daoquan ZHANG ; Wenfang CHENG ; Liuqin JIANG ; Xiqiao ZHOU
Chinese Journal of Gastroenterology 2020;25(10):601-605
Background: The occurrence of gastrointestinal symptoms in cirrhotic patients with gastroesophageal varices (GOV) after endoscopic treatment is obvious, and the role of gastric myoelectrical activity (GMA) and autonomic nerve function imbalance in the development of gastrointestinal symptoms has not been clarified. Aims: To investigate the changes of GMA and autonomic nerve function in cirrhotic patients with GOV after endoscopic treatment. Methods: Twenty-five cirrhotic patients with GOV from May 2019 to October 2019 at the First Affiliated Hospital of Nanjing Medical University were enrolled, and 10 patients with gastric polyp were served as controls. Electrogastrogram (EGG) and heart rate variability (HRV) were detected before the operation, 1 day after the operation and 5 days after the operation in GOV group. For the gastric polyp group, EGG and HRV were detected before the operation and 1 day after the operation. Changes of GMA and autonomic nerve function were compared between the two groups. Results: No significant differences in GMA and autonomic nerve function were found between GOV group and gastric polyp group before and 1 day after the operation (P>0.05). LF, LF/HF were significantly increased 1 day after the operation in GOV group (P<0.05), while HF was significantly decreased (P<0.01), and the percentage of bradygastria was significantly increased (P<0.01). There were no significant differences in GMA and autonomic nerve function between pre-operation and 5 days after the operation in GOV group (P>0.05). There were no significant differences in GMA and autonomic nerve function 1 day after the operation compared with pre-operation in gastric polyp group (P>0.05). Compared with Child-Pugh A group, Child-Pugh B group had more obvious GMA abnormalities, and the difference was statistically significant (P<0.05). Conclusions: In cirrhotic patients with GOV, the percentage of bradygastria and the sympathetic activity increased, and the vagal activity decreased 1 day after the operation. These results suggest that GMA and autonomic nerve dysfunction may be related to the gastrointestinal symptoms after endoscopic treatment.
6.Transition of autophagy and apoptosis in fibroblasts depends on dominant expression of HIF-1α or p53.
Min LI ; Yidan SU ; Xiaoyuan GAO ; Jiarong YU ; Zhiyong WANG ; Xiqiao WANG
Journal of Zhejiang University. Science. B 2022;23(3):204-217
It has been revealed that hypoxia is dynamic in hypertrophic scars; therefore, we considered that it may have different effects on hypoxia-inducible factor-1α (HIF-1α) and p53 expression. Herein, we aimed to confirm the presence of a teeterboard-like conversion between HIF-1α and p53, which is correlated with scar formation and regression. Thus, we obtained samples of normal skin and hypertrophic scars to identify the differences in HIF-1α and autophagy using immunohistochemistry and transmission electron microscopy. In addition, we used moderate hypoxia in vitro to simulate the proliferative scar, and silenced HIF-1α or p53 gene expression or triggered overexpression to investigate the changes of HIF-1α and p53 expression, autophagy, apoptosis, and cell proliferation under this condition. HIF-1α, p53, and autophagy-related proteins were assayed using western blotting and immunofluorescence, whereas apoptosis was detected using flow cytometry analysis, and cell proliferation was detected using cell counting kit-8 (CCK-8) and 5-bromo-2'-deoxyuridine (BrdU) staining. Furthermore, immunoprecipitation was performed to verify the binding of HIF-1α and p53 to transcription cofactor p300. Our results demonstrated that, in scar tissue, HIF-1α expression increased in parallel with autophagosome formation. Under hypoxia, HIF-1α expression and autophagy were upregulated, whereas p53 expression and apoptosis were downregulated in vitro. HIF-1α knockdown downregulated autophagy, proliferation, and p300-bound HIF-1α, and upregulated p53 expression, apoptosis, and p300-bound p53. Meanwhile, p53 knockdown induced the opposite effects and enhanced HIF-1α, whereas p53 overexpression resulted in the same effects and reduced HIF-1α. Our results suggest a teeterboard-like conversion between HIF-1α and p53, which is linked with scar hyperplasia and regression.
Apoptosis
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Autophagy
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Cell Hypoxia
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Fibroblasts/metabolism*
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit/metabolism*
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Tumor Suppressor Protein p53/metabolism*
7.Safety comparison of high-dose methotrexate via different intravenous infusion devices in pediatric patients with malignant brain tumors
Xiqiao XU ; Zhengyuan SHI ; Chunjing YANG ; Shumei WANG
China Pharmacy 2023;34(7):844-848
OBJECTIVE To compare the safety of high-dose methotrexate (HD-MTX) via peripherally inserted central catheter (PICC) and totally implantable venous access port (TIVAP) in pediatric patients with malignant brain tumors. METHODS Patients with malignant brain tumors who received HD-MTX via PICCs or TIVAPs in our hospital from July 2018 to April 2022 were retrospectively analyzed. Clinical data were collected to compare differences in blood concentration of methotrexate (MTX),the incidence of adverse events (including adverse drug reactions and catheter-related complications) and length of stay in hospital. Multivariate linear regression was applied to analyze the factors that influenced the blood concentration of MTX. RESULTS A total of 107 patients were included in the study,with 65 patients in the PICC group and 42 patients in the TIVAP group. Blood concentration of MTX at 24 h (C24 h) in TIVAP group was significantly higher than PICC group ([ 126.87±61.99) μmol/L vs. (102.45±48.77) μmol/L,P<0.05). There was no significant difference in blood concentration of MTX at 42 h (C42 h),compared with PICC group (P>0.05). Results of multivariate linear regression analysis showed that TIVAP was associated with the increase of C24 h(P<0.05). No significant differences were observed in the incidence of adverse events and the length of stay in the hospital between 2 groups (P>0.05). CONCLUSIONS Risk of adverse events is not increased,although the MTX C24 h level is elevated after administration of TIVAP. TIVAP is a safe choice for HD-MTX therapy with implementing therapeutic drug monitoring.
8.Associations of MTRR gene polymorphism and methotrexate plasma concentration and adverse drug reaction in children with intracranial tumors
Danqi ZHAO ; Miao LI ; Zhengyuan SHI ; Xiqiao XU ; Shumei WANG
China Pharmacy 2024;35(21):2646-2651
OBJECTIVE To investigate the impact of the methionine synthase reductase (MTRR) rs10380 C>T gene polymorphism on methotrexate (MTX) plasma concentration, adverse drug reaction, and prognosis in children with intracranial tumors. METHODS Peripheral blood was collected from children with intracranial tumors, and genomic DNA was extracted. The MTRR rs10380 C>T genotype was analyzed using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. The association of the MTRR rs10380 C>T gene polymorphism with the ratio of MTX plasma concentration to dose (C/D ratio), adverse drug reaction, tumor recurrence, and metastasis was analyzed. Bioinformatics analysis was used to explore the association of the rs10380 genotype and MTRR gene expression and its possible mechanisms. RESULTS A total of 75 children were included in the study. The distribution frequencies of the wild-type CC genotype and C allele of rs10380 were 62.67% and 81.33%, respectively, while the distribution frequencies of the variant CT genotype and T allele were 37.33% and 18.67%, respectively, which were in accordance with Hardy-Weinberg equilibrium(P>0.05). The incidence of electrolyte disorders (51.06%) and tumor metastasis rate (57.45%) in children with the CC genotype were significantly higher than those with the CT genotype (P<0.05). No significant differences were observed in the 24-hour and 42-hour C/D ratios and recurrence rates between the two genotypes of children (P>0.05). Bioinformatics analysis showed that MTRR protein mainly works in conjunction with 10 proteins, including MMAA, and was involved in various biological processes such as sulfur amino acid biosynthesis. CONCLUSIONS The MTRR rs10380 CC genotype may be a risk factor for electrolyte disorders and tumor metastasis in children with intracranial tumors after MTX chemotherapy.