OBJECTIVES: To reprogram human placental fibroblasts (HPFs) into chemically induced epithelioid-like cells (ciEP-Ls) using a combination of small-molecule compounds. METHODS: HPFs cultured under normoxic conditions were identified using immunofluorescence assay, PCR and chromosomal karyotyping. Under hypoxic conditions (37 ℃, 5% O₂), HPFs were cultured in a medium containing small-molecule compounds C6TSEDRVAJZ (CHIR99021, 616452, TTNPB, SAG, EPZ5676, DZNep, Ruxolitinib, VTP50469, Afuresertib, JNK-IN-8, and EZM0414), and the cell morphology was observed daily. The expression levels of epithelial cell markers in the induced cells were detected by immunofluorescence, Western blotting and PCR. Chromosomal karyotyping of the induced cells was performed and the induction efficiency was calculated. RESULTS: Before induction, HPFs showed positive expressions of fibroblast surface markers CD34 and vimentin and were negative for epithelial surface markers. PCR results showed high expressions of fibroblast-specific genes S100A4 and COL1A1 in HPFs with a normal human diploid karyotype. After one day of induction, the HPFs underwent morphological changes from a multinodular spindle shape to a round or polygonal shape, which was morphologically characteristic of ciEP-Ls. On day 4 of induction, the cells exhibited high expressions of the epithelial cell markers E-cadherin and Lin28A. RT-qPCR results also showed that the cells expressed the epithelial markers Smad3, GLi3, PAX8, WT1, KRT19, and KRT18 with significantly down-regulated expressions of all the fibroblast surface markers and a normal human diploid karyotype. The reprogramming efficiency of HPFs into ciEP-Ls ranged from (64.53±2.8)% to (68.10±3.6)%. CONCLUSIONS The small-molecule compound combination C6TSEDRVAJZ is capable of inducing HPFs into ciEP-Ls under hypoxic conditions with a high induction efficiency.
Humans ; Fibroblasts/drug effects* ; Pregnancy ; Female ; Cell Differentiation/drug effects* ; Pyrimidines/pharmacology* ; Placenta/cytology* ; Cells, Cultured ; Pyridines/pharmacology* ; Pyrazoles/pharmacology* ; Epithelial Cells/cytology*

Objective:To investigate a surgical method and clinical effect on reconstruction of infective ulcer and soft tissue defects in dorsal fingers of the elderly patients with free perforator flap of peroneal artery.Methods:From March 2016 to June 2022, 13 elderly patients with infective ulcer and soft tissue defects in dorsal fingers were reconstructed with free perforator flaps of peroneal artery. The patients were 65-70 years with an average age of 66.5 years. Cause of infection: 10 ulceration and soft tissue defects were caused by diabetes and 3 by injury. Seven infective ulceration and soft tissue defects were in dorsal index fingers, 3 in dorsal middle fingers and 3 in dorsal ring fingers with the size of soft tissue defects at 2.0 cm×4.5 cm-2.0 cm×5.5 cm with an exposure of tendon and phalange. The donor site of the flaps was of contralateral calf and the flaps were 2.5 cm×5.0 cm-2.5 cm×6.0 cm in size. All donor sites were sutured directly. All patients were included in the postoperative follow-up at outpatient clinic to observe the appearance and sensation of the flap as well as finger movement.Results:All flaps survived and all wounds achieved stage I healing, without recurrence of infection. Twelve patients had the postoperative follow-up for 12 to 27 months, with an average of 21.6 months. There were satisfactory appearance of flaps and the function of fingers. Sensation of flaps recovered to S 2 in 5 patients and S 3 in 7 patients. The recovery of hand function was evaluated according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, with 8 hands in excellent and 4 in good. Conclusion:The free perforator flap of peroneal artery has advantages of constant vascular anatomy, reliable blood supply, moderate thickness and direct closure of donor site. It is a useful clinical method in reconstruction of infective ulcer and soft tissue defects in dorsal fingers of the elderly patients.

Objective To investigate the effects and underlying mechanisms of a spray prepared from exosomes derived from M2 macrophages induced by interleukin-4 （IL-4） and tantalum particles （Ta） on the healing of pressure ulcers. Methods Bone marrow-derived macrophages were polarized into M2 macrophages using IL-4 or Ta, and exosomes （Exo-IL-4/Exo-Ta） were extracted. The regulatory effects of Exo-IL-4/Exo-Ta on M1 macrophage phenotypes and fibroblast matrix secretion were evaluated in vitro. Proteomic analysis was conducted to explore the biological processes and regulatory networks associated with Exo-Ta. A rat pressure ulcer model was used to assess the effects of Exo-IL-4/Exo-Ta spray on wound healing rate, inflammatory cell infiltration, and collagen deposition. Results In vitro, Exo-IL-4/Exo-Ta induced the polarization of M1 macrophages to M2 macrophages, reduced the secretion of pro-inflammatory factors, and promoted the expression of anti-inflammatory substances. Additionally, Exo-IL-4/Exo-Ta enhanced the production of collagen and fibronectin in fibroblasts. Proteomic analysis revealed that Exo-Ta primarily participated in biological processes such as energy metabolism and macromolecule biosynthesis. In vivo, Exo-IL-4/Exo-Ta spray accelerated wound healing, reduced inflammatory infiltration, and improved tissue remodeling in the rat pressure ulcer model. Conclusion Exosome sprays derived from M2 macrophages could accelerate pressure ulcer healing by modulating inflammation and promoting tissue regeneration, which demonstrated excellent clinical application potential.

Objective:To investigate a surgical method and clinical effect on reconstruction of infective ulcer and soft tissue defects in dorsal fingers of the elderly patients with free perforator flap of peroneal artery.Methods:From March 2016 to June 2022, 13 elderly patients with infective ulcer and soft tissue defects in dorsal fingers were reconstructed with free perforator flaps of peroneal artery. The patients were 65-70 years with an average age of 66.5 years. Cause of infection: 10 ulceration and soft tissue defects were caused by diabetes and 3 by injury. Seven infective ulceration and soft tissue defects were in dorsal index fingers, 3 in dorsal middle fingers and 3 in dorsal ring fingers with the size of soft tissue defects at 2.0 cm×4.5 cm-2.0 cm×5.5 cm with an exposure of tendon and phalange. The donor site of the flaps was of contralateral calf and the flaps were 2.5 cm×5.0 cm-2.5 cm×6.0 cm in size. All donor sites were sutured directly. All patients were included in the postoperative follow-up at outpatient clinic to observe the appearance and sensation of the flap as well as finger movement.Results:All flaps survived and all wounds achieved stage I healing, without recurrence of infection. Twelve patients had the postoperative follow-up for 12 to 27 months, with an average of 21.6 months. There were satisfactory appearance of flaps and the function of fingers. Sensation of flaps recovered to S 2 in 5 patients and S 3 in 7 patients. The recovery of hand function was evaluated according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, with 8 hands in excellent and 4 in good. Conclusion:The free perforator flap of peroneal artery has advantages of constant vascular anatomy, reliable blood supply, moderate thickness and direct closure of donor site. It is a useful clinical method in reconstruction of infective ulcer and soft tissue defects in dorsal fingers of the elderly patients.

Objective:To compare the effects of different positioning and puncture methods for epidural anesthesia in obese parturients undergoing cesarean section.Methods:In this prospective, randomized, controlled study, 175 obese parturients, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with a body mass index of > 30 kg/m 2, who underwent elective cesarean section with epidural anesthesia at Henan Provincial People′s Hospital from July 2023 to April 2024, were selected and assigned into 4 groups using a random number table method: palpation positioning + median approach to puncture group (PM group, n=44), body surface marker positioning + paramedian approach to puncture group (BP group, n=43), ultrasound-assisted positioning + paramedian approach to puncture group (UP group, n=45), and ultrasound-assisted positioning + ultrasound-guided in-plane approach to puncture group (UU group, n=43). The procedures were performed by anesthesiology residents skilled in the four different epidural anesthesia positioning and puncture methods. Main observation indicators: success rate of puncture at first attempt and total success rate of puncture. Secondary observation indicators: the number of puncture, changes in puncture interspaces, positioning time, puncture time, anesthesia operation time, postoperative nausea and vomiting, sensory abnormalities at the puncture site, bleeding at the puncture site, maternal satisfaction rates, and occurrence of low back pain at day 7, 1 month, 2 months, and 3 months postoperatively. Results:In this study, 2 cases were excluded from PM group, 2 from BP group, 2 from UP group, and 2 from UU group. The total success rates of puncture were as follows: 95% (40/42) in PM group, 98% (40/41) in BP group, 93% (40/43) in UP group, and 98% (40/41) in UU group. There was no significant difference in the total success rates of puncture among the four groups ( P>0.05). Compared to PM group, the success rate of puncture at first attempt was significantly increased, the rate of changes in puncture interspaces was decreased, the positioning time was shortened, the incidence of nausea and vomiting was decreased, satisfaction rates were increased, and the incidence of low back pain was reduced at 7 days after operation in BP group; the success rate of puncture at first attempt was significantly increased, the number of puncture was reduced, the rate of changes in puncture interspaces was decreased, the positioning time, puncture time and anesthesia operation time were shortened, the satisfaction rates were increased, and the incidence of low back pain was decreased at 7 days after operation in UP group; the rate of changes in puncture interspaces was significantly decreased, and the positioning time was shortened in UU group ( P<0.05). Compared to BP group, the puncture time was significantly shortened in UP group, and the number of puncture was significantly increased, the anesthesia operation time was prolonged, the incidence of nausea and vomiting was increased, and the satisfaction rate was decreased in UU group ( P<0.05). Compared to UP group, the success rate of puncture at first attempt was significantly decreased, the number of puncture was increased, the puncture time and anesthesia operation time were prolonged, the incidence of nausea and vomiting was increased, and the satisfaction rates were decreased in UU group ( P<0.05). Conclusions:Ultrasound-assisted positioning combined with the paramedian approach to puncture can raise the success rate of puncture at first attempt, reduce the development of puncture-related complications, and increase the maternal satisfaction in obese parturients undergoing cesarean section.

Objective To assess the predictive value of bedside diaphragmatic ultrasound in predicting pulmonary complications(PPCs)after thoracoscopic lobectomy.Methods The patients who had undergone elective thoracoscopic lobectomy in Henan Provincial People's Hospital were collected.General information and perioperative indicators were recorded.Diaphragmatic motility was detected by bedside ultrasound preoperatively and on postopera-tive days 1,3,and 5.The patients were divided into two groups(PPCs group and non-PPCs group).Statistically different parameters in univariate analysis were included in multivariate logistic regression analysis to screen inde-pendent influencing factors of PPCs.Receiver operating curve(ROC)was drawn,and the performance of diaphragm ultrasound for predicting PPCs was evaluated by the area under the curve(AUC).Results 949 patients were included in this study.PPCs occurred in 537 patients(57.5% ).Univariate analysis showed that as compared with the non-PPCs group,the proportion of diabetic patients and postoperative VAS score in the PPCs group increased,time to chest tube removal and one-lung ventilation,and postoperative hospital stay were longer.PPCs group had a lower DE value on the operative side and healthy side on postoperative days 1 and 3 and on the operative side alone on day 5(P<0.05).Multivariate logistic regression analysis showed that prolonged one-lung ventilation time,diabetes mellitus,DE values on the operative side and unoperative side on postoperative days 1 and 3,and decreased DE value on the operative side on day 5 were independent risk factors for PPCs.The ROC curve shows that the efficacy of DE for pre-dicting PPCs on the operative and healthy sides on postoperative days 1 and 3 was relatively high(AUC of 0.797,0.821,0.933,and 0.929;respectively).The efficacy of DE for predicting PPCs on the operative side was poor on postoperative day 5(AUC of 0.703 and 0.512,respectively).Conclusions A decrease in postoperative DE,prolonged one-lung ventilation time,and diabetes are independent risk factors for PPCs after thoracoscopic lobectomy.Bedside ultrasound evaluation of DE has a higher value in predicting PPCs three days after surgery.

Objective:To measure the overall and lobulated volume of the liver with different degrees of liver fibrosis and to further observe pathological changes such as liver microvasculature, hepatocyte apoptosis, and regeneration in order to understand the macroscopic volume changes of the liver during liver fibrosis and its relationship with liver tissue microscopic pathology in patients with chronic liver disease.Methods:53 patients with chronic hepatitis B, alcoholic fatty liver disease, autoimmune liver disease, nonalcoholic fatty liver disease, and drug-induced chronic liver disease who underwent both liver biopsy tissue and abdominal magnetic resonance imaging were collected. Patients were divided into early (F1-2), middle (F3-4), and late (F5-6) in accordance with the Ishak fibrosis stage and Masson stain. The liver and spleen volumes were measured using ITK-SNAP software. CD31 immunohistochemical staining was used to reflect intrahepatic angiogenesis. Ki67 and HNF-4α multiplex immunohistochemical staining were used to reflect hepatocyte regeneration. GS staining was used to determine parenchymal extinction lesions. TUNEL staining was used to observe hepatocyte apoptosis. Spearman correlation analysis was used to analyze the relationship between liver volume changes and liver histopathological changes.Results:As liver fibrosis progressed, the total liver volume and right lobe liver volume gradually decreased ( P<0.05), while the spleen volume gradually increased ( P<0.05). The expression of CD31 and GS gradually increased ( P<0.05), and the expression of Ki67 first increased and then decreased ( P<0.05). The positivity rate of CD31 was negatively correlated with the right lobe liver volume ( r=-0.609, P<0.001) and the total liver volume ( r=-0.363, P=0.017). The positivity rate of Ki67 was positively correlated with the right lobe liver volume ( r=0.423, P=0.018), while the positivity rate of apoptotic cells was significantly negatively correlated with the total liver volume ( r=-0.860, P<0.001). The positivity rate of GS was negatively correlated with the right lobe liver volume ( r=-0.440, P=0.002), and the number of PELs was negatively correlated with RV ( r=-0.476, P=0.013). The CD31 positive staining area was negatively correlated with the Ki67 positive staining area( r=-0.511, P=0.009). Conclusion:As liver fibrosis progresses, patients with chronic liver disease have a depletion in total liver volume and right lobe liver volume, and this is mainly in correlation with fewer liver cells and liver tissue microvasculature disorders.

Objective:To explore the application effect of preoperative autologous blood localization method in laparoscopic resection of gastric stromal tumors in unfavorable areas of the stomach.Methods:A retrospective analysis was conducted on the case data of 40 patients with gastric stromal tumors in unfavorable locations admitted to Jinjiang Hospital from January 2019 to December 2022. The patients were divided into a control group (intraoperative endoscopic localization method) and an autologous blood localization group according to different intraoperative lesion localization methods, with 20 cases in each group. The surgical time, intraoperative blood loss, hospitalization time, postoperative exhaust time, and adverse reactions were compared between the two groups.Results:The surgery time of the autologous blood localization group was shorter than that of the control group [(92.30±8.80)min vs (108.20±14.87)min, P<0.05]. There was no statistically significant difference in intraoperative bleeding, hospitalization time, and postoperative exhaust time between the two groups (all P>0.05). Two groups of patients did not show an increase in inflammatory indicators such as white blood cells and C-reactive protein on the day after surgery. Both groups of patients did not experience adverse reactions such as fever, abdominal pain, or postoperative complications. Conclusions:The autologous blood injection localization method provides a safe, simple, and effective method for preoperative localization of gastric stromal tumors in unfavorable areas of the stomach under laparoscopy, and is worthy of clinical promotion and use.

Objective:To compare the effects of different positioning and puncture methods for epidural anesthesia in obese parturients undergoing cesarean section.Methods:In this prospective, randomized, controlled study, 175 obese parturients, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, with a body mass index of > 30 kg/m 2, who underwent elective cesarean section with epidural anesthesia at Henan Provincial People′s Hospital from July 2023 to April 2024, were selected and assigned into 4 groups using a random number table method: palpation positioning + median approach to puncture group (PM group, n=44), body surface marker positioning + paramedian approach to puncture group (BP group, n=43), ultrasound-assisted positioning + paramedian approach to puncture group (UP group, n=45), and ultrasound-assisted positioning + ultrasound-guided in-plane approach to puncture group (UU group, n=43). The procedures were performed by anesthesiology residents skilled in the four different epidural anesthesia positioning and puncture methods. Main observation indicators: success rate of puncture at first attempt and total success rate of puncture. Secondary observation indicators: the number of puncture, changes in puncture interspaces, positioning time, puncture time, anesthesia operation time, postoperative nausea and vomiting, sensory abnormalities at the puncture site, bleeding at the puncture site, maternal satisfaction rates, and occurrence of low back pain at day 7, 1 month, 2 months, and 3 months postoperatively. Results:In this study, 2 cases were excluded from PM group, 2 from BP group, 2 from UP group, and 2 from UU group. The total success rates of puncture were as follows: 95% (40/42) in PM group, 98% (40/41) in BP group, 93% (40/43) in UP group, and 98% (40/41) in UU group. There was no significant difference in the total success rates of puncture among the four groups ( P>0.05). Compared to PM group, the success rate of puncture at first attempt was significantly increased, the rate of changes in puncture interspaces was decreased, the positioning time was shortened, the incidence of nausea and vomiting was decreased, satisfaction rates were increased, and the incidence of low back pain was reduced at 7 days after operation in BP group; the success rate of puncture at first attempt was significantly increased, the number of puncture was reduced, the rate of changes in puncture interspaces was decreased, the positioning time, puncture time and anesthesia operation time were shortened, the satisfaction rates were increased, and the incidence of low back pain was decreased at 7 days after operation in UP group; the rate of changes in puncture interspaces was significantly decreased, and the positioning time was shortened in UU group ( P<0.05). Compared to BP group, the puncture time was significantly shortened in UP group, and the number of puncture was significantly increased, the anesthesia operation time was prolonged, the incidence of nausea and vomiting was increased, and the satisfaction rate was decreased in UU group ( P<0.05). Compared to UP group, the success rate of puncture at first attempt was significantly decreased, the number of puncture was increased, the puncture time and anesthesia operation time were prolonged, the incidence of nausea and vomiting was increased, and the satisfaction rates were decreased in UU group ( P<0.05). Conclusions:Ultrasound-assisted positioning combined with the paramedian approach to puncture can raise the success rate of puncture at first attempt, reduce the development of puncture-related complications, and increase the maternal satisfaction in obese parturients undergoing cesarean section.

Objective:To explore the clinical effect of cosmetic reconstruction for partial defect of distal segment of digits.Methods:Form January 2018 to January 2021, the Department of Hand Surgery of Institute for Hygiene of Ordnance Industry(The 521 Hospital of Weapon Industry) admitted 129 patients with partial defect of distal segment of thumb or fingers with phalange or tendon exposure. The patients were 111 males and 18 females with an average age of 34(17-59) years old. The sizes of nailbed defect were 0.4 cm×1.1 cm-1.8 cm×2.0 cm, the length of phalange defect was 0.4-1.8 cm, and the sizes of the soft tissue defect were 1.6 cm×1.8 cm-3.2 cm×4.8 cm. Great toe tissue flaps were used to reconstruct the partial defect of distal segment of thumb or fingers after debridement. Wounds of fibular flap of great toe in 77 cases were directly sutured in 17 patients. The donor sites in rest 60 great toes were narrowed first and then repaired with skin grafts in 10 cases, with artificial dermis in 28 cases and with transverse V-Y advancement flaps of ipsilateral great toes for 22 cases. Forty-nine of 52 donor site wounds for hallux toenail flap were repaired with artificial dermis and 3 with free peroneal artery perforator flaps. The method was outpatient follow-up. Postoperative follow-up lasted until July 2022. The check-items for follow-up included: occurrence of necrosis, appearance, shape and texture of the flap, appearance of the reconstructed nails, TPD of the reconstructed digit pulps, tolerance to cold on the scars of flaps, flexion and extension of the reconstructed digits. The healing time of phalanges of the reconstructed digits was evaluated by X-rays. The appearance, sensation, the tolerance to cold of great toe and the movement of donor foot were also assessed.Results:Postoperative follow-up lasted for 18 to 24 months, with an average of 21 months. A total of 128 flaps survived. Necrosis occurred in 1 fibular hallux flap, the necrosis was cured with a reverse digital proper artery island flap. Thereafter, all flaps healed well. The appearance, shape, texture and nails of reconstructed digits were close to the contralateral digits. The reconstructed thumb and finger were evaluated according to Zook, 127 cases were excellent and 2 cases were good.TPD of the pulps of the reconstructed digits was 4-10 mm. The mean score of the Vancouver scar scale(VSS) was 0.6 for scars of the reconstructed digits. The mean score of the Visual analog scale(VAS) was 0.3 for the tolerance to cold. Flexion and extension function recovered well in all the reconstructed digits. According to the Evaluation Criteria of Upper limb Function Hand Surgery Society of Chinese Medical Association, the function of hand was excellent in 127 cases and 1 in each of good and fair. X-rays of all digits showed the phalange healing of the reconstructed digits, with an average healing time of 2 months after surgery, without phalange resorption, infection, nonunion nor stress fracture. There was no difference between the length of the donor great toe and the contralateral toe, except the donor site of the great toe nail root. There was no significant visual difference between the appearance of the donor great toe and the contralateral toe. TPD of the pulps of donor great toe was 4-8 mm. The mean score of the VSS was 1.4 for scars in the donor great toe. The mean score of the VAS was 0.7 for the tolerance to cold of the donor great toe. There was no stress fracture at donor site, and the functions of donor foot were not affected when walking, running, jumping and tiptoeing in all patients.Conclusion:It is an ideal method for reconstructing a partial defect of distal segment of digit by great toe flap. It can not only reconstruct the partial defect of distal segment of the digit, but also results in a good appearance and satisfactory functions of the reconstructed digit. Damage to the donor site is minimum. The length of the donor great toe is unchanged, and it has little impact on sensation and appearance of the donor great toe. Meanwhile, there is no adverse effect on walking, running and jumping with the donor foot.