Lingguizhugan Decoction (LGZG) demonstrates significant efficacy in treating various cardiovascular diseases clinically, yet its precise mechanism of action remains elusive. This study aimed to elucidate the potential mechanisms and effects of LGZG on isoproterenol (ISO) continuous stimulation-induced chronic heart failure (CHF) in mice, providing direct experimental evidence for further clinical applications. In vivo, continuous ISO infusion was administered to mice, and ventricular myocytes were utilized to explore LGZG?s potential mechanism of action on the ?1-adrenergic receptor (?1-AR)/Gs/G protein-coupled receptor kinases (GRKs)/?-arrestin signaling deflection system in the heart. The findings reveal that LGZG significantly reduced the messenger ribonucleic acid (mRNA) expression of hypertrophy-related biomarkers [atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP)] and improved cardiac remodeling and left ventricular diastolic function in mice with ISO-induced CHF. Furthermore, LGZG inhibited the overactivation of Gs/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling and downregulated the downstream transcriptional activity of cAMP-response element binding protein (CREB) and the expression of the coactivator CBP/P300. Notably, LGZG downregulated the expression of ?-arrestin1 and GRK 2/3/5 while upregulating the expression of ?1-AR and ?-arrestin2. These results suggest that LGZG inhibits Gs/cAMP/PKA signaling and ?-arrestin/GRK-mediated desensitization and internalization of ?1-AR, potentially exerting cardioprotective effects through the synergistic regulation of the ?1-AR/Gs/GRKs/?-arrestin signaling deflection system via multiple pathways.
Animals ; Heart Failure/genetics* ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Male ; G-Protein-Coupled Receptor Kinases/genetics* ; Mice, Inbred C57BL ; Humans ; Isoproterenol ; Arrestins/genetics* ; Chronic Disease

Objective The high post-surgery recurrence rate of endometriosis(EMs)has emerged as a challenge in the long-term manaagement of the condition.This study is aimed at investigating the mechanisms of Neiyiting(NYT)decoction in preventing postoperative recurrence of EMs.Methods An animal model of EMs postoperative recurrence and a model of endometrial stromal cells(hEM15A)cocultured with macrophages(RAW 264.7 cell line)were established for both in vivo and in vitro experiments.An autotransplantation method was used to establish a rat model of EMs.The rats were divided into 4 groups(6 rats per group)and received the corresponding treatments:a Model group receiving distilled water,a Gestrinone group receiving gestrinone at 0.325 mg/kg,a low-dose NYT(NYT-L)group receiving NYT decoction at 5.04 g/(kg-d),and a high-dose NYT(NYT-H)group receiving NYT decoction at 10.08 g/(kg-d).The treatment was administered for 3 weeks via intragastric gavage.In addition,6 SD rats were randomly selected for the control group(Control group),and were given distilled water for 3 weeks via intragastric gavage.The sizes and pathological changes of recurrent lesions in EMs rats were observed.Immunohistochemistry and qRT-PCR were performed to assess the expression of M1 macrophage marker CD86 protein and mRNA in vivo.Additionally,immunohistochemistry and qRT-PCR were used to assess the expression of indicator proteins related to the triggering receptor expressed on myeloid cells 1(TREM1)/Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)signaling pathway and mRNA.The proliferation of hEM15A cells in the coculture experiment was observed.Flow cytometry was performed to determine the polarization of RAW264.7 macrophages,and qRT-PCR was used to determine the expression levels of inducible nitric oxide synthase(iNOS)and interleukin 1β(IL-1β)mRNA.Western blot was performed to determine the expression of signaling pathway-related indicator proteins in vitro.ELISA was performed to determine the levels of inflammatory factors in vitro.Results Compared with the Model group,the volume of recurrent lesions in the NYT-H group was reduced(P＜0.01).Findings from the macrophage M1 polarization assessment showed that the expression levels of CD86 protein and mRNA in the recurrent lesions of the Model group were higher than those in the control group(P＜0.01).The expression levels of CD86 protein and mRNA in the recurrent lesions of the NYT-H group were lower than those of the Model group(P＜0.01).In addition,the RAW 264.7 cell experiment further verified that NYT decoction could reduce the number of CD86-positive macrophages induced by plasmids overexpressing TREM1 and reduce the expression of IL-1β and iNOS mRNA(P＜0.01).The results of the hEM15A cell proliferation assay showed that NYT decoction down-regulated KI-67 protein expression in hEM15A cells induced by macrophage M1 polarization(P＜0.01).The results of TREM1/TLR4/NF-κB signaling pathway showed that the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the Model group were higher than those of the control group(P＜0.01).Compared with those in the Model group,the protein and mRNA expression levels of TREM1,TLR4,and NF-κB in the recurrent lesions of the NYT-H group were lower(P＜0.01).In addition,the coculture experiment of RAW264.7 and hEM15A cells further confirmed that NYT decoction reduced the expression of TREM1,TLR4,and P-P65 proteins(P＜0.01).Conclusion NYT decoction can inhibit macrophage M1 polarization through the TREM1/TLR4/NF-κB signaling pathway,improve the inflammation level,and inhibit the formation of ectopic endometrial lesions,thereby preventing postoperative recurrence of EMs.

OBJECTIVE To observe the clinical efficacy and effect on serum thymic stromal lymphopoietin(TSLP)levels of pa-tients with advanced liver cancer of qi deficiency and toxic stasis type by Jiawei Yupingfeng San.METHODS Using random double blind method,120 patients with advanced liver cancer of qi deficiency and toxic stasis type were randomly divided into 3 groups:Jiawei Yupingfeng San group,Yupingfeng San group,and placebo group,each consisting of 40 cases.All patients in the 3 groups were given conventional treatment such as radiotherapy,chemotherapy,interventional or targeted therapy;Jiawei Yupingfeng San group was given Jiawei Yupingfeng San granules,Yupingfeng San group was given Yupingfeng San granules,and placebo group was given placebo.The course of treatment was 2 months.The changes of Karnofsky functional status score(KPS score),TCM syndrome score,tumor size and serum TSLP level in the 3 groups were observed before and after treatment,and the correlation between the changes of tumor size and TSLP was analyzed.RESULTS After treatment,the KPS scores of Yupingfeng San group and Jiawei Yupingfeng San group were sig-nificantly increased(P<0.05,P<0.01),TCM syndrome score were decreased(P<0.01),tumor growth(P<0.05,P<0.01)was de-layed,and serum TSLP levels(P<0.05,P<0.01)were decreased.Furthermore,there was a slight positive correlation between chan-ges in tumor size and changes in TSLP(P<0.05).In terms of improving tumor size,the curative effect of Jiawei Yupingfeng San group was better than that of Yupingfeng San group(P<0.05).During the treatment period,no obvious adverse reactions were observed in the 3 groups of patients.CONCLUSION Combined with conventional treatment,Jiawei Yupingfeng San can significantly delay tumor growth in patients with advanced liver cancer of qi deficiency and toxic stasis type and improve patients'TCM syndromes and their qual-ity of survival.The therapeutic mechanism is related to reducing the expression of serum TSLP and improving the immune status of pa-tients,thereby delaying the growth of tumors.

Objective:To evaluate the efficacy and safety of modified mushroom-shaped occluder in the treatment of refractory thoracogastric-airway fistulas.Methods:From March 1, 2022 to June 30, 2023, 12 patients with refractory thoracogastric-airway fistulas underwent the placement of modified mushroom-shaped occluder at the Department of Gastroenterology, the First Affiliated Hospital of Nanjing Medical University were enrolled. The baseline clinical data of patients such as gender, age, course of disease, and fistula diameter were recorded. The data of operation and follow-up, such as operation time and method, intraoperative and postoperative complications were also collected. The occlusion efficacy at 1 month and 6 months after surgery, as well as the improvement of body mass index (BMI) and scores of the short form 36 (SF-36) were analyzed. Paired t test and non-parametric test were used for statistical analysis. Results:There were 10 males and 2 females among the 12 patients. The median age was 66.5 years old (ranged from 53.0 to 69.0 years old), the median course of disease was 7.5 months (ranged from 3.0 to 39.0 months), and the diameter of fistula was (9.3±3.4) mm. The occluder placements were successful in all the 12 patients, with 6 cases intracavitary release and 6 extracavitary release. The operation time was (30.9±9.9) min and the time of occluder placement was (3.5±1.3) min. One patient had minor (<2 mL) bleeding during the operation and 2 patients reported mild foreign body sensation but tolerable after operation. All patients resumed oral feeding and nasojejunal tubes were removed before discharge. The follow-up time of 12 patients was (11.3±1.7) months. The initial effective occlusion rate was 11/12, and the complete occlusion rate was 9/12. Two patients died but neither were related to the procedure or instruments. The BMI of 12 patients at 1 month after surgery was (18.5±1.9) kg/m 2, which was higher than that before operation ((17.6±2.3) kg/m 2), the BMI at 6 months after operation was (20.3±2.5) kg/m 2, which was higher than that at 1 month after operation, and the differences were statistically significant ( t=-4.15 and -4.45, P=0.002 and 0.001). The scores of 8 domains of SF-36 including physical functioning, general health, vitality, mental health, role-physical, bodily pain, social functioning and role-emotional of 12 patients before operation, at 1 month after operation and 6 months after operation were 49.6±13.6, 63.3±13.5 and 75.4±8.6, 17.1±11.2, 33.2±14.5 and 56.0±12.2, 30.0±12.6, 45.0±13.5 and 67.5±8.7, 41.3±18.7, 52.0±15.4 and 68.0±8.2, 0.0 (0.0 to 75.0), 25.0 (0.0 to 100.0) and 50.0 (25.0 to 100.0), 87.8 (44.0 to 100.0), 90.8 (57.0 to 100.0) and 100.0 (94.0 to 100.0), 12.5 (0.0 to 50.0), 50.0 (37.5 to 75.0) and 81.3 (50.0 to 87.5), 0.0 (0.0 to 100.0), 66.7 (33.3 to 100.0) and 100.0 (33.3 to 100.0), respectively. The scores of 8 domains at 1 month after operation were all higher than those before operation, and the differences were statistically significant ( t=-5.25, -5.32, -4.87 and -2.51, Z=-2.97, -2.20, -3.11 and -3.00; all P<0.05). The scores of 8 domains at 6 months after operation were all higher than those at 1 month after operation, and the differences were statistically significant ( t=-4.34, -7.48, -7.10 and -4.64, Z=-2.49, -2.20, -2.97 and -2.07; all P<0.05). Conclusion:The clinical application of the improved mushroom-shaped occluder in the treatment of refractory thoracogastric-airway fistulas is effective and relatively safe.

Objective:To evaluate the safety and efficacy of endoscopic injection of autologous platelet rich plasma(PRP)in the treatment of refractory upper gastrointestinal fistula(RUGF).Methods:From June 1, 2022 to March 1, 2023, at the Department of Gastroenterology of the First Affiliated Hospital of Nanjing Medical University, 8 RUGF patients who received PRP treatment were enrolled. The baseline data such as gender, age, course of disease, and body mass index, etc. as well as the PRP treatment-related data such as the time of first treatment, length of hospital stay after the first treatment, and adverse events were recorded. The postoperative follow-up lasted till 6 months after the first treatment to assess fistula healing, and to record body mass index and health survey short form. Paired t-test and non-parametric test were used for statistical analysis. Results:Among the 8 RUGF patients, there were 7 males and 1 female. The median age was 58.5 years old (ranged from 27.0 to 75.0 years old), and the median duration of the disease was 14 months (ranged from 4 to 120 months). The maximum diameter of the fistula was(4.50±2.62) mm, the concentration multiple of PRP was (4.02±0.48) times. The operation time of the first endoscopic PRP treatment was (21.88±4.52) min; the median length of the first postoperative hospital stay was 2 days (ranged from 1 to 2 days), and the median total number of treatment was 4 (ranged from 1 to 5). The healing time was (2.57±1.72) months, 7 RUGF patients healed within 6 months after the first treatment, and the fistula was narrowed in 1 patient. There were no adverse events during or after the operation. After 6 months of treatment, the body mass index and the scores of 8 dimensions of the health survey short form which included physical functioning, physical role functioning, body pain, general health, vitality, social functioning, emotional role functioning and mental health were all higher than those before treatment ((20.91±2.15) kg/m 2 vs. (18.67±3.21) kg/m 2, 86.88±13.35 vs. 58.13±20.34, 100 (0 to 100) vs. 0(0 to 100), 83.06±11.74 vs. 56.94±28.86, 67.88±26.77 vs. 31.88±13.08, 81.88±13.87 vs. 46.25±24.02, 76.56±22.60 vs. 37.50±26.73, 100 (0 to 100) vs. 0 (0 to 100), 78.00±17.37 vs. 51.50±22.77), and the differences were statistically significant ( t=-3.40 and -3.87, Z=2.06, t=-3.03, -4.26, -4.73 and -6.06, Z=-2.06, t=-4.32; P=0.012, 0.006, 0.039, 0.019, 0.004, 0.002, 0.001, 0.039 and 0.003). Conclusion:Endoscopic injection of autologous PRP for the treatment of RUGF is safe and effective, and has a significant promotion effect on the healing of small (maximum diameter ≤ 6 mm) fistula.

Objective:To evaluate the safety and clinical efficacy of the novel double disc-shaped gastrointestinal occluder (hereinafter referred to as occluder) in treatment of refractory tracheoesophageal fistula (TEF).Methods:From July 1, 2020 to January 31, 2021, 10 patients with refractory TEF treated with occluder at Department of Gastroenterology, the First Affiliated Hospital with Nanjing Medical University were collected. The patients′ clinical data such as gender, age, body mass index and fistula diameter were recorded. The success rate of operation, intraoperative and postoperative complications, operation time, postoperative hospital stay, efficacy of fistula closure and postoperative recovery were analyzed. The Karnofsky score and body mass index of patients 1 month and 3 months after operation were compared with those before operation for curative effect evaluation. Paired t test was used for statistical analysis. Results:Among the 10 TEF patients, there were 7 males and 3 females, the median age was 64.5 years old (ranged from 49.0 to 78.0 years old), the body mass index was (18.0±2.5) kg/m 2 and the diameter of the fistula was (1.2±0.6) cm. Occluder placement was successful in all patients. The operation time was (17.6±7.8) min (ranged from 7.0 to 30.0 min). Two cases had little bleeding during the operation, and there was no bleeding after the operation. The postoperative hospital stay was (5.9±4.0) d (ranged from 1.0 to 12.0 d). Among the 10 TEF patients, fistula of 5 patients were completely blocked, 4 cases were partially blocked and 1 case was ineffectively blocked, the effective rate of blocking was 9/10. One month follow-up after operation showed that the symptoms of choking and coughing during eating were significantly improved in 9 patients, and the symptoms of choking and coughing during eating were significantly improved in 1 patient after waist diameter of 12 mm occluder was replaced with the occluder of 15 mm. The 3-month follow-up after operation showed that the occluders were in the right place in 8 patients, the occluder was displaced in 1 patient and the occluder was removed and treated with enteral nutrition. One patient died due to the recurrence of esophageal cancer. The Karnofsky score of TEF patients 3 months after operation and the body mass index of TEF patients 1 month and 3 months after operation were higher than those before operation (70.0±34.0 vs. 46.0±10.7, (19.32±2.59) and (19.73±2.92) kg/m 2 vs. (18.03±2.50) kg/m 2), and the differences were statistically significant ( t=-3.09, -2.37 and -2.82, all P<0.05). Conclusions:Gastrointestinal occluder is safe and effective in the treatment of refractory TEF.

To evaluate the therapeutic effectiveness and safety of a novel gastrointestinal occluder device for gastrobronchial fistula. Data of 5 patients diagnosed as having gastrobronchial fistula who received treatment by a novel gastrointestinal occluder device at the First Affiliated Hospital of Nanjing Medical University from July to August 2020 were retrospectively analyzed. The total operation time, occluding time, intraoperative and postoperative complications, postoperative hospital stay and patients′ satisfaction were reviewed. Regular follow-up was conducted, and the short-term curative effect of occluding was evaluated 1 month after operation.All patients were males with age of 58-69 years. The course of fistula ranged 3-16 months and the diameter ranged 0.3-1.0 cm. All 5 patients achieved technical success with operation time of 38-88 minutes and occluding time of 8-24 minutes. The postoperative hospital stay ranged 3-5 days and the patients′ satisfaction score was 10. No severe complications occurred during or after operation. One month after endoscopic therapy, fistula was completely occluded in 4 patients. One patient died due to severe pulmonary infection and multiple organ failure although the bucking symptom after drinking and eating recovered before. Endoscopic closure of gastrointestinal fistula by means of the novel gastrointestinal occluder device is safe and effective.

Objective To establish an ELISA for quantitative determination of decoy receptor 3 (DcR3) in human plasma.Methods A solid phase double antibody sandwich method was established for quantitative determination of DcR3.The anti-DcR3 antibody was immobilized onto ELISA plate.DcR3 in samples was captured by anti-DcR3 on ELISA plate and then detected by biotin-anti-DcR3 and subsequent peroxidase-labeled streptavidin,and the color was developed by adding substrate.The standard DcR3 samples on the same plate were detected simultaneously to calculate the DcR3 concentrations in unknown samples.The sensitivity,specificity,precision,recovery,linearity and DcR3 range in normal human adults were assessed.Results The sensitivity of the developed assay was 0.051 ng/mL.The intra-coefficient of variation (CV) was less than 10％ and inter-CV was less than 15％.The average recovery rate was 90.50％.When 2-fold amount of anti-TNF-α was added into the coated antibodies,10-fold amount of biotin-labeled anti-LIGHT,antiFAS or anti-TNF-α was added into the detection antibodies,or 10 fold amount of purified LIGHT protein was added into the standard DcR3 samples as competitor,no disturbing effects on standard curve were found.The linear range of the assay was from 0.25 to 16 ng/mL (r≥0.98).The concentration of DcR3 tested in 128 plasma samples from healthy adults was (0.21 ± 0.05) ng/mL with 95％ CI ranged from 0.14 to 0.28 ng/mL and no difference of age and sex was found.Conclusion The established ELiSA for determining plasma DcR3 exhibited high specificity,sensitivity,precision,fine linearity and wide detecting range.This method could be used for quantification of DcR3 in plasma.

Aim To observe the correlation between the TPL's suppression on myofibrolbasts(MFBs)activation and TGF-β1/ERK/Smad3 pathway by performing in vivo and in vitro experiments.Methods In vitro model of MFBs activation was set up by stimulating fibroblasts with TGF-β1,and in vivo model of MFBs activation in radiated lung tissue was built by thoracic radiation on C57BL/6 mice.MFBs activation was analyzed by detecting the expression of α-SMA(using RT-PCR and Western blot)and Col Ⅰ(using RT-PCR and ELISA methods).The levels of p-ERK,p-Smad3(Ser208)and p-Smad3(Ser423)were measured by Western blot.ERK siRNA and Smad3 siRNA were used to observe the status of ERK and Smad3 in MFBs activation.Results TGF-β1 activated p-ERK/p-Smad3(Ser208)and p-Smad3(Ser423),increased the expression of α-SMA and synthesis of Col Ⅰ,which indicated MFBs activation.siRNA knockdown assay showed that both ERK and Smad3 were involved in regulating the levels of α-SMA and Col Ⅰ,and ERK influenced MFBs transformation possibly through its phosphorylation of Smad3(Ser208).TPL treatment inhibited the phosphorylation activation of ERK,Smad3(Ser208),Smad3(Ser423)in vitro and in vivo,therefore significantly reduced the level of α-SMA and Col Ⅰ,and the number of activated MFBs was decreased.Conclusion TPL mitigates radiation-induced pulmonary fibrosis by inhibiting the activation of MFBs,which is partly through suppressing TGF-β1/ERK/Smad3 pathway.

Objective:To observe the clinical efficacy of Chinese medical gargle on gingivitis of the patients wearing fixed appliance. Methods:60 gingivitis patients with fixed orthodontic treatment were randomly divided into chlorhexidine gargle group( CG group) and Chinese medical gargle group(CMG group)(n=30). Gingival index(GI) and sulcus bleeding index(SBI) in all patients were tested, then the patients were treated by scaling and curettage, and guided to use gargling with corresponding gargle 3 times a day for 14 d. GI and SBI were tested after 7 and 14 days respectively. Data were analysed. Results:After 7 d treatment, GI and SBI were decreased( P<0. 05) and there was no obvious difference between the 2 groups(P<0. 05). After 14 d treatment GI and SBI in CMG group de-creased more than those in CG group(P<0. 05). Conclusion:Chinese medical gargle is effective in the treatment of gingivitis of the patients during fixed orthodontic treatment.