Objective To investigate the effects of in vitro culture with different concentrations of glucose and serum on hypoxia-induced activation and Yes-associated protein(YAP)expression in primary cardiac fibroblasts(CF).Methods Primary CF were isolated from ICR suckling mice with enzyme digestion and then purified using differential adhesion technique.The cells were ran-domly assigned into 6 groups:low glucose high serum group(1.0 g/L glucose+10％serum),high glucose high serum group(4.5 g/L glucose+10％serum),low glucose medium serum group(1.0 g/L glucose+5％serum),high glucose medium serum group(4.5 g/L glucose+5％serum),low glucose serum-free group(1.0 g/L glucose+0％serum)and high glucose serum-free group(4.5 g/L glucose+0％serum).In 24 h after above culture,the cell morphology was observed.Western blotting was used to detect the expression of α-smooth muscle actin(α-SMA),transforming growth factor beta(TGF-β),type Ⅰ collagen(Col Ⅰ).and YAP.Immunofluorescence assay was em-ployed to observe the expression and localization of α-SMA and YAP.EDU staining and cell scratch assay were applied to measure cell proliferation and migration ability,respectively.Results After culture in 1％O2 for 24 h,the cell morphology was gradually altered toward myofibroblasts as the concentration of glucose rose and the serum level fell in the culture medium,especially in the group of high glucose and serum-free.The expression levels of Col Ⅰ,TGF-β,α-SMA and YAP were significantly higher in the high glucose no serum group than in the high glucose high serum group(P＜0.05).The fluorescence intensity of α-SMA was obviously increased in the low glucose no serum group than the low glucose high serum group[(9.23±2.45)％vs(2.40±2.04)％,P＜0.05].Compared to the low-sugar serum-free group,the high glucose serum-free group showed a significant increase in the levels of Col Ⅰ,TGF-β,α-SMA and YAP(P＜0.05).Notably higherα-SMA fluorescence intensity,more YAP nuclear translocation,and enhanced cell migration were observed in the high glucose serum-free group when compared to the high glucose high serum group and the low glucose serum-free group(P＜0.05).Conclusion Under the hypoxic condition of 1％O2,high-glucose serum-free is the most appropriate culture condition to construct an in vitro cell model of hypoxia-induced activation,and the activation of YAP is of most significant im-portance.

Objective:To discuss the expression of Rh family C glycoprotein(RHCG)in the esophageal squamous cell carcinoma(ESCC)tissue and its effect on the malignant biological behavior of ESCC cells,and to clarify the value of RHCG as a diagnostic and prognostic marker for the ESCC patients.Methods:A total of 143 ESCC tissue samples and 105 adjacent normal tissue samples were collected.Using immunohistochemical staining method,141 ESCC samples were divided into two groups:RHCG low expression group(immunohistochemistry score≤6)and RHCG high expression group(immunohistochemistry score>6).Immunohistochemical method was used to detect the RHCG protein expression in 143 ESCC tissues and 105 normal tissues,and the relationship between the clinicopathological characteristics of the ESCC patients was analyzed.Receiver operating characteristic(ROC)curve and Kaplan-Meier survival analysis were used to evaluate the value of RHCG in diagnosis and prognosis of the ESCC patients;univariate and multivariate COX regression analysis were used to determine the independent risk factors affecting the prognosis of the ESCC patients.Gene Expression Profiling Interactive Analysis(GEPIA2)database was used to analyze the expression of RHCG mRNA in various tumor tissues.The ESCC TE-1 cells were cultured and transfected in to 6-well cell culture plates with different Lipofectamine2000∶RHCG ratios;the cells in RHCG transfection group were transfected with weights of 2.0,2.5,and 3.0 μg for 24 and 48 h,respectively,and the cells in NC group transfected with empty vector as control.Western blotting method was used to detect the RHCG protein expression level in the TE-1 cells in various groups after transfection at different concentrations and verify the optimal transfection conditions;cell counting kit-8(CCK-8)assay was used to detect the proliferation activities of the TE-1 cells;plate clone formation assay was used to detect the colony formation numbers of the TE-1 cells;Transwell chamber assay was used to detect the numbers of migrating TE-1 cells.Results:Compared with adjacent normal tissue,the RHCG gene expression level in various cancer tissues including ESCC,glioblastoma multiforme,and head and neck squamous cell carcinoma was significantly decreased(P<0.05).RHCG protein was mainly located on the cell membrane of normal esophageal squamous epithelial cells;the RHCG protein expression intensity in ESCC tissues was lower than that in adjacent normal esophageal tissue(χ2=109.373,P<0.001),and the patients in RHCG low expression group had poorer differentiation than those in RHCG high expression group(P=0.041).The area under the curve(AUC)value of RHCG for diagnosing ESCC was 0.86,with sensitivity and specificity of 95.1%and 75.0%,respectively;the Kaplan-Meier survival analysis results showed that compared with high RHCG expression group,the patients in low RHCG expression group had shorter survival time and poorer prognosis[harard ratio(HR)=0.269,95%confidence interval(CI):0.113-0.639,P=0.020];the COX regression analysis results showed that low RHCG expression could serve as an independent risk factor affecting the prognosis of ESCC[HR=4.569,95%CI=1.315-15.877,P=0.017)].The Western blotting results verified that the optimal transfection condition was 3.0 μg RHCG plasmid for 48 h,at which time RHCG overexpression was optimal and RHCG protein expression level was highest.The CCK-8 assay results showed that compared with control group,the proliferation activity in RHCG overexpression group was decreased on the 4th day after cell seeding(P<0.001).In the TE-1 cells,the colony formation number of the TE-1 cells in RHCG over-expression group was lower than that in control group(t=17.70,P<0.001).The Transwell chamber assay results showed that compared with control group,the number of migrating cells in RHCG over-expression group was decreased(t=23.74,P<0.001).Conclusion:RHCG expression is decreased in ESCC tissues and associated with poor prognosis in ESCC patients;overexpression of RHCG can inhibit the proliferation and migration of the TE-1 cells,providing a theoretical basis for RHCG as a novel diagnostic and prognostic marker and therapeutic target for ESCC.

Objective:To evaluate the clinical efficacy of Cai's gynecological nourishing kidney therapy for patients with diminished ovarian reserve (DOR).Methods:A randomly controlled trail was conducted. A total of 63 patients with DOR who were treated in the gynecology outpatient clinic of Yueyang Hospital of Integrated Traditional Chinese and Western Medicine affiliated to Shanghai University of Traditional Chinese Medicine from September 2020 to January 2022 were set as observation subjects, and were divided into control group ( n=31) and observation group ( n=32) according to the random number table method. The patients in the control group were treated with Femoston, and the patients in the observation group were treated with Cai's gynecological nourishing kidney therapy on the basis of Femoston treatment. Both groups were treated for 3 menstrual cycles. Both antral follicle count (AFC) was performed by vaginal ultrasound, serum levels of TGF-β1, IL-1β, IL-2, IL-6 and IL-21 were detected by ELISA, and serum levels of FSH, LH, E2, anti-Müllerian hormone (AMH) and MDA were detected by biochemical methods. The TCM syndrome score was used to evaluate the severity of symptoms, and the menstrual score was used to evaluate the abnormal menstruation. Clinical efficacy was evaluated. Results:The total effective rate was 87.50% (28/32) in the observation group and 58.06% (18/31) in the control group, with statistical significance ( χ2=8.42, P=0.004). After treatment, the AFC in the observation group (3.93±2.32 vs. 2.21±2.18, t=3.03) was higher than that of the control group ( P<0.01). After treatment, the serum FSH level of the observation group [10.05 (8.35, 21.48) IU/L vs. 20.60 (8.00, 43.30) IU/L, Z=2.18] was lower than that of the control group ( P<0.05), and the level of AMH [0.19 (0.03, 0.47) μg/L vs. 0.02 (0.01, 0.24) μg/L, Z=-1.54] in the observation group was higher than that of the control group ( P<0.05). The serum levels of TGF-β1 [(68.27±11.76) ng/L vs. (55.33±13.03) ng/L, t=4.14] and IL-1β [(58.00±7.53) ng/L vs. (52.31±8.06) ng/L, t=2.89] in the observation group were higher than those in the control group, and the levels of IL-6 [(33.26±7.45) ng/L vs. (40.69±11.69) ng/L, t=3.02], the level of IL-21 [(118.37±15.56) ng/L vs. (140.43±25.51) ng/L, t=-4.04] was lower than that of the control group ( P<0.01). Conclusions:Cai's gynecological nourishing kidney therapy combined with Fenmaotong can effectively increase the AFC of DOR patients and improve clinical efficacy. Its mechanism of action may be related to reducing serum FSH and LH levels and increasing E2 levels.

Objective To investigate the effects of in vitro culture with different concentrations of glucose and serum on hypoxia-induced activation and Yes-associated protein(YAP)expression in primary cardiac fibroblasts(CF).Methods Primary CF were isolated from ICR suckling mice with enzyme digestion and then purified using differential adhesion technique.The cells were ran-domly assigned into 6 groups:low glucose high serum group(1.0 g/L glucose+10％serum),high glucose high serum group(4.5 g/L glucose+10％serum),low glucose medium serum group(1.0 g/L glucose+5％serum),high glucose medium serum group(4.5 g/L glucose+5％serum),low glucose serum-free group(1.0 g/L glucose+0％serum)and high glucose serum-free group(4.5 g/L glucose+0％serum).In 24 h after above culture,the cell morphology was observed.Western blotting was used to detect the expression of α-smooth muscle actin(α-SMA),transforming growth factor beta(TGF-β),type Ⅰ collagen(Col Ⅰ).and YAP.Immunofluorescence assay was em-ployed to observe the expression and localization of α-SMA and YAP.EDU staining and cell scratch assay were applied to measure cell proliferation and migration ability,respectively.Results After culture in 1％O2 for 24 h,the cell morphology was gradually altered toward myofibroblasts as the concentration of glucose rose and the serum level fell in the culture medium,especially in the group of high glucose and serum-free.The expression levels of Col Ⅰ,TGF-β,α-SMA and YAP were significantly higher in the high glucose no serum group than in the high glucose high serum group(P＜0.05).The fluorescence intensity of α-SMA was obviously increased in the low glucose no serum group than the low glucose high serum group[(9.23±2.45)％vs(2.40±2.04)％,P＜0.05].Compared to the low-sugar serum-free group,the high glucose serum-free group showed a significant increase in the levels of Col Ⅰ,TGF-β,α-SMA and YAP(P＜0.05).Notably higherα-SMA fluorescence intensity,more YAP nuclear translocation,and enhanced cell migration were observed in the high glucose serum-free group when compared to the high glucose high serum group and the low glucose serum-free group(P＜0.05).Conclusion Under the hypoxic condition of 1％O2,high-glucose serum-free is the most appropriate culture condition to construct an in vitro cell model of hypoxia-induced activation,and the activation of YAP is of most significant im-portance.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective:To compare the efficacy of biplanar fixation combined with bone grafting and proximal femoral nail anti-rotation (PFNA) in the treatment of critically complicated osteoporotic intertrochanteric femoral fracture .Methods:A retrospective cohort study was conducted to analyze the clinical data of 28 patients with critically complicated osteoporotic intertrochanteric femoral fracture, admitted to Union Hospital, Tongji Medical College of Huazhong University of Science and Technology from January 2020 and December 2022, including 3 males and 25 females, aged 70-91 years [(79.4±6.3)years]. T score for bone mineral density was -2.5~-4.1 SD［（-3.3±0.6）SD］. All the patients were found with type A2.2-A3.3 fracture based on AO classification, and were complicated with trochanteric lateral wall fracture. Among them, 16 patients underwent biplanar fixation combined with bone grafting (biplanar fixation group), while 12 underwent PFNA internal fixation (PFNA group). All the patients received anti-osteoporosis therapy after surgery. The two groups were compared in terms of the operative time, intraoperative blood loss, hemoglobin levels at 3 days postoperatively, and time to weight-bearing. The visual analogue scale (VAS) scores and Harris hip scores at 1, 3, 6 months postoperatively, and at the last follow-up and the incidence of complications were also detected in the two groups.Results:All the patients were followed up for 12-16 months [(14.1±1.4)months]. In the biplanar fixation group, the operative time was (75.1±15.3)minutes, significantly longer than (45.6±14.2)minutes in the PFNA group ( P<0.01); the intraoperative blood loss was (234.1±11.8)ml, significantly more than (170.0±13.4)ml in the PFNA group ( P<0.01); the hemoglobin level at 3 days postoperatively was (82.6±9.3)g/L in the biplanar fixation group, higher than (64.8±6.8)g/L in the PFNA group ( P<0.01). The time to weight-bearing was (1.1±0.7)weeks in the biplanar fixation group, significantly shorter than (3.2±1.2)weeks in the PFNA group ( P<0.01). There were no statistically significant differences between the two groups in VAS scores and Harris hip scores at 1, 3, 6 months postoperatively, and at the last follow-up ( P>0.05). The VAS scores and Harris hip scores in the two groups were gradually improved with the prolongation of postoperative time ( P<0.05 or 0.01). No complications such as neurovascular injuries were observed in either group. One patient in the biplanar fixation group developed lower extremity deep vein thrombosis (DVT), with a complication rate of 6.3%, while 2 patients in the PFNA group developed lower extremity DVT and 4 hypostatic pneumonia, with a complication rate of 50.0% ( P<0.05). Conclusion:Compared with PFNA internal fixation, biplanar fixation with bone grafting has the advantages of less postoperative blood loss, earlier weight-bearing exercises and lower incidence of complications in the treatment of critically complicated osteoporotic intertrochanteric femoral fracture, despite longer operative time and more intraoperative blood loss.

Postoperative infection of internal fixation of closed fractures the lower limbs in adults represents a devastating complication, characterized by diagnostic challenges, prolonged treatment duration and high disability rates. Current management of these infections faces multiple challenges, such as difficulties in early accurate diagnosis, and various controversies about the treatment plan, leading to poor overall diagnosis and treatment results. To address these issues, based on evidence-based medicine and principles with emphasis on scientific rigor, clinical applicability and innovation, the Trauma Branch of the Chinese Medical Association, Orthopedic Branch of the Chinese Medical Doctor Association, Orthopedics Branch of the Chinese Medical Association, and Trauma Orthopedics and Polytrauma Group of the Resuscitation and Emergency Committee of the Chinese Medical Doctor Association have collaboratively organized a panel of relevant experts to develop the Guideline for diagnosis and treatment of infection after internal fixation of closed lower limb fractures in adults ( version 2025). The guideline proposed 10 recommendations, aiming to provide a foundation for standardized diagnosis and treatment of postoperative infection in adults with closed lower limb fractures.

Objective:To compare the efficacy of biplanar fixation combined with bone grafting and proximal femoral nail anti-rotation (PFNA) in the treatment of critically complicated osteoporotic intertrochanteric femoral fracture .Methods:A retrospective cohort study was conducted to analyze the clinical data of 28 patients with critically complicated osteoporotic intertrochanteric femoral fracture, admitted to Union Hospital, Tongji Medical College of Huazhong University of Science and Technology from January 2020 and December 2022, including 3 males and 25 females, aged 70-91 years [(79.4±6.3)years]. T score for bone mineral density was -2.5~-4.1 SD［（-3.3±0.6）SD］. All the patients were found with type A2.2-A3.3 fracture based on AO classification, and were complicated with trochanteric lateral wall fracture. Among them, 16 patients underwent biplanar fixation combined with bone grafting (biplanar fixation group), while 12 underwent PFNA internal fixation (PFNA group). All the patients received anti-osteoporosis therapy after surgery. The two groups were compared in terms of the operative time, intraoperative blood loss, hemoglobin levels at 3 days postoperatively, and time to weight-bearing. The visual analogue scale (VAS) scores and Harris hip scores at 1, 3, 6 months postoperatively, and at the last follow-up and the incidence of complications were also detected in the two groups.Results:All the patients were followed up for 12-16 months [(14.1±1.4)months]. In the biplanar fixation group, the operative time was (75.1±15.3)minutes, significantly longer than (45.6±14.2)minutes in the PFNA group ( P<0.01); the intraoperative blood loss was (234.1±11.8)ml, significantly more than (170.0±13.4)ml in the PFNA group ( P<0.01); the hemoglobin level at 3 days postoperatively was (82.6±9.3)g/L in the biplanar fixation group, higher than (64.8±6.8)g/L in the PFNA group ( P<0.01). The time to weight-bearing was (1.1±0.7)weeks in the biplanar fixation group, significantly shorter than (3.2±1.2)weeks in the PFNA group ( P<0.01). There were no statistically significant differences between the two groups in VAS scores and Harris hip scores at 1, 3, 6 months postoperatively, and at the last follow-up ( P>0.05). The VAS scores and Harris hip scores in the two groups were gradually improved with the prolongation of postoperative time ( P<0.05 or 0.01). No complications such as neurovascular injuries were observed in either group. One patient in the biplanar fixation group developed lower extremity deep vein thrombosis (DVT), with a complication rate of 6.3%, while 2 patients in the PFNA group developed lower extremity DVT and 4 hypostatic pneumonia, with a complication rate of 50.0% ( P<0.05). Conclusion:Compared with PFNA internal fixation, biplanar fixation with bone grafting has the advantages of less postoperative blood loss, earlier weight-bearing exercises and lower incidence of complications in the treatment of critically complicated osteoporotic intertrochanteric femoral fracture, despite longer operative time and more intraoperative blood loss.

Pulmonary vein stenosis (PVS) is an extremely rare and lethal disease caused by multiple etiologies. PVS has a bimodal distribution in the population, affecting children and adults. Congenital PVS is the usual PVS type in children, which sometimes develops after cardiothoracic surgery. Acquired PVS, in turn, is the most common PVS type in adults. A review of the relevant literature has shown that PVS after radiofrequency ablation of atrial fibrillation is the most common, as well as that caused by compression of proliferative fibrous tissues or tumor in the mediastinum (eg, PVS caused by fibrosing mediastinitis, lung tumors, metastases, etc). This article provides a comprehensive review of PVS in terms of embryology and anatomy, etiology and triggers, classification, clinical symptoms and signs, treatment, and prognosis, intending to promote the understanding and treatment of this disease.

Objective:To explore the clinical effects of Wenjing Decoction in treating endometriosis syndrome with cold coagulation and blood stasis.Methods:A randomized controlled trial was performed. A total of 90 cases were collected and with distributed random number method of drugs were divided into control group and the treatment group with 45 cases each. The treatment group was given Wenjing Decoction granules, while control group was given placebo (10% Wenjing Decoction). Both groups were taken for 14 consecutive days from the first day of menstruation as a course of 1, a total of 3 courses. Before and after treatment, main observation indexes including dysmenorrhea, Traditional Chinese Medicine (TCM) syndromes and the secondary outcome including scores of menstruation, short form McGill pain questionnaire (SF-MPQ) were detected by subjective evaluation, interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α), and prostaglandin F2α (PGF2α) level in human serum tissue were evaluated by enzyme linked immunosorbent assay and the clinical and TCM syndromes specific effects were determined.Results:Totally 7 patients were lost, including 1 case in the treatment group and 6 cases in control group. The number of actual observation cases was 83 (44 cases in the treatment group and 39 cases in control group). The clinical effective rate in the treatment group [75.00% (33/44)] was statistically higher than that in control group [23.08% (9/39), P<0.001]. As for therapeutic effect of TCM syndromes, the effective rate in the treatment group [70.45% (31/44)] was obviously higher than that in control group [20.51% (8/39), P<0.001]. After treatment, compared with the results of control group, menstruation [2.00 (1.00,4.00)], dysmenorrhea [7.25 (5.00,9.88)], SF-MPQ [10.00 (4.50,18.00)], TCM syndromes [13.50 (9.25, 20.00)] and the data of IL-8 [9.80 (5.73,27.53) ng/L], TNF-α [5.05 (4.10,6.20) ng/L] and PGF2α [(147.16±22.24) ng/L] in the treatment group were decreased obviously [4.00 (2.00,6.00), P=0.022; 8.00 (6.50,12.00), P=0.030; 13.00 (4.00,24.00), P<0.001; 17.00 (9.00, 26.00), P<0.001; 18.20 (10.00,30.90) ng/L, P=0.013; 5.80 (4.50,10.70) ng/L, P=0.035;(165.17±30.33) ng/L, P=0.001]. Conclusion:Wenjing Decoction can treat TCM syndromes of endometriosis patients with cold coagulation and blood stasis, and the clinical curative effect was significant. The mechanism may involve enhancing immunity and relieving uterine smooth muscle contraction.