Objective We aimed to evaluate the efficacy and safety of Shengxuebao Mixture in the treatment of cancer-related anemia(CRA)presenting with syndrome of deficiency of liver and kidney combined with syndrome of deficiency of both qi and blood.Methods A randomized,double-blind,placebo-controlled,multicenter clinical trial was conducted.Eligible patients with malignant tumors meeting the inclusion and exclusion criteria were enrolled from 26 hospitals,including Dongzhimen Hospital,Beijing University of Chinese Medicine,Xiaogan Central Hospital,and Yangzhou Hospital of Traditional Chinese Medicine,from June 1,2022,to September 30,2024.Patients were allocated 1:1 to either the experimental group receiving Shengxuebao Mixture or the control group receiving its simulator(placebo)using a block randomization method under double-blind conditions.Both groups received 15 mL orally three times daily for 28 consecutive days.The primary efficacy indicators included the hemoglobin(Hb)improvement rate(RHb)and the traditional Chinese medicine(TCM)syndrome improvement rate(RTCM)at week 4 of treatment.The secondary efficacy indicators encompassed Hb and red blood cell(RBC)count,Karnofsky Performance Status(KPS)score,TCM syndrome score,individual TCM symptom scores,and changes in each of these indicators compared to the baseline period at weeks 2,4,and 6 of treatment.Safety evaluations were conducted at week 4 of treatment.Results A total of 239 patients were enrolled,with 225 cases included in the Full Analysis Set(FAS)(109 in the experimental group vs.116 control group),163 in the Per Protocol Set(PPS)(77 vs.86),and 225 in the Safety Set(SS)(109 vs.116).Baseline characteristics between groups showed no significant differences.Significant differences were observed between the experimental and control groups in RHb at week 4(FAS:49.51%vs.35.24%,P<0.05;PPS:53.25%vs.36.05%,P<0.05)and RTCM at week 4(FAS:61.54%vs.39.62%,P<0.01;PPS:64.94%vs.40.70%,P<0.01).At weeks 2,4,and 6,the experimental group showed greater improvements in Hb and RBC counts than the control group.Additionally,the TCM syndrome scores were lower in the experimental group than in the control group at these time points.Except for week 2 in PPS,the KPS improvement was better in the experimental group than in the control group(P<0.05).The experimental group also demonstrated a greater reduction in scores for individual TCM symptoms such as spiritlessness and weakness,poor appetite and reduced food intake at weeks 4 and 6 compared to the control group(P<0.05,P<0.01).Furthermore,the reduction in vertigo score was more pronounced in the experimental group at week 6(P<0.01).For the score of pale and lusterless complexion,only in the PPS was the reduction from baseline more significant in the experimental group than in the control group at weeks 4 and 6(P<0.05).No significant differences were observed between the experimental and control groups in the incidence of all adverse events or drug-related adverse reactions.Conclusion Shengxuebao Mixture demonstrates significant efficacy in patients with CRA presenting syndrome of deficiency of liver and kidney combined with syndrome of deficiency of both qi and blood,effectively increasing Hb levels,ameliorating TCM syndromes,alleviating clinical symptoms,and enhancing functional status,with no significant difference in adverse drug reactions compared to the placebo.

Objective We aimed to evaluate the efficacy and safety of Shengxuebao Mixture in the treatment of cancer-related anemia(CRA)presenting with syndrome of deficiency of liver and kidney combined with syndrome of deficiency of both qi and blood.Methods A randomized,double-blind,placebo-controlled,multicenter clinical trial was conducted.Eligible patients with malignant tumors meeting the inclusion and exclusion criteria were enrolled from 26 hospitals,including Dongzhimen Hospital,Beijing University of Chinese Medicine,Xiaogan Central Hospital,and Yangzhou Hospital of Traditional Chinese Medicine,from June 1,2022,to September 30,2024.Patients were allocated 1:1 to either the experimental group receiving Shengxuebao Mixture or the control group receiving its simulator(placebo)using a block randomization method under double-blind conditions.Both groups received 15 mL orally three times daily for 28 consecutive days.The primary efficacy indicators included the hemoglobin(Hb)improvement rate(RHb)and the traditional Chinese medicine(TCM)syndrome improvement rate(RTCM)at week 4 of treatment.The secondary efficacy indicators encompassed Hb and red blood cell(RBC)count,Karnofsky Performance Status(KPS)score,TCM syndrome score,individual TCM symptom scores,and changes in each of these indicators compared to the baseline period at weeks 2,4,and 6 of treatment.Safety evaluations were conducted at week 4 of treatment.Results A total of 239 patients were enrolled,with 225 cases included in the Full Analysis Set(FAS)(109 in the experimental group vs.116 control group),163 in the Per Protocol Set(PPS)(77 vs.86),and 225 in the Safety Set(SS)(109 vs.116).Baseline characteristics between groups showed no significant differences.Significant differences were observed between the experimental and control groups in RHb at week 4(FAS:49.51%vs.35.24%,P<0.05;PPS:53.25%vs.36.05%,P<0.05)and RTCM at week 4(FAS:61.54%vs.39.62%,P<0.01;PPS:64.94%vs.40.70%,P<0.01).At weeks 2,4,and 6,the experimental group showed greater improvements in Hb and RBC counts than the control group.Additionally,the TCM syndrome scores were lower in the experimental group than in the control group at these time points.Except for week 2 in PPS,the KPS improvement was better in the experimental group than in the control group(P<0.05).The experimental group also demonstrated a greater reduction in scores for individual TCM symptoms such as spiritlessness and weakness,poor appetite and reduced food intake at weeks 4 and 6 compared to the control group(P<0.05,P<0.01).Furthermore,the reduction in vertigo score was more pronounced in the experimental group at week 6(P<0.01).For the score of pale and lusterless complexion,only in the PPS was the reduction from baseline more significant in the experimental group than in the control group at weeks 4 and 6(P<0.05).No significant differences were observed between the experimental and control groups in the incidence of all adverse events or drug-related adverse reactions.Conclusion Shengxuebao Mixture demonstrates significant efficacy in patients with CRA presenting syndrome of deficiency of liver and kidney combined with syndrome of deficiency of both qi and blood,effectively increasing Hb levels,ameliorating TCM syndromes,alleviating clinical symptoms,and enhancing functional status,with no significant difference in adverse drug reactions compared to the placebo.

Objective:To investigate the protective effect and possible mechanism of Tangshenbao on renal damage in diabetic nephropathy (DN) rats.Methods:Totally 36 SPF male SD rats were randomly divided into normal group ( n=6) and model group ( n=30). The DN rat model was prepared by single high-dose intraperitoneal injection of STZ. According to the random number table method, the rats were divided into model group, irbesartan group and Tangshenbao low-, medium- and high-dosage groups, with 6 rats in each group. Drug intervention lasted for 8 weeks. The general condition and body weight of rats in each group were recorded. The blood glucose, kidney index, 24 h urine protein (24 h UTP), SCr and BUN levels were detected. The pathological morphology of renal tissue was observed by PAS staining and transmission electron microscopy. The mRNA and protein expressions of Ets-1, TGF-β1, Smad2 and Smad3 in renal tissue were detected by real-time fluorescence quantitative PCR and Western blot. Results:Compared with model group, the body weight of Tangshenbao low, medium and high dose groups and irbesartan group significantly increased ( P<0.01). The kidney index decreased ( P<0.05 or P<0.01). The contents of 24 hUTP, BUN and SCr significantly decreased ( P<0.05 or P<0.01). Glomerular volume was significantly reduced ( P<0.05 or P<0.01), the mRNA expressions of Ets-1 (1.59 ± 0.06, 1.47 ± 0.04, 1.31 ± 0.03, 1.39 ± 0.03 vs. 1.64 ± 0.04), TGF-β1 (1.65 ± 0.05, 1.59 ± 0.03, 1.38 ± 0.05, 1.49 ± 0.04 vs. 1.77 ± 0.08), Smad2 (1.48 ± 0.05,1.39 ± 0.05, 1.22 ± 0.03, 1.31 ± 0.04 vs. 1.54 ± 0.05), Smad3 (1.57 ± 0.04, 1.48 ± 0.03, 1.28 ± 0.03, 1.39 ± 0.02 vs. 1.64 ± 0.05) in renal tissue of rats significantly decreased ( P<0.05 or P<0.01), the protein expressions of Ets-1 (1.33 ± 0.32, 1.16 ± 0.38, 0.77 ± 0.06, 0.84 ± 0.06 vs. 1.97 ± 0.43), TGF-β1 ( 1.35 ± 0.14, 1.24 ± 0.22, 0.94 ± 0.13, 1.07 ± 0.06 vs. 1.63 ± 0.20), Smad2 (1.24 ± 0.26, 1.14 ± 0.31, 0.77 ± 0.28, 0.85 ± 0.19 vs. 1.72 ± 0.34) and Smad3 (1.29 ± 0.14, 1.19 ± 0.21, 0.85 ± 0.39, 0.90 ± 0.37 vs. 1.76 ± 0.21) decreased ( P<0.05 or P<0.01). Conclusion:Tangshenbao can improve renal damage in DN rats, and its mechanism may be related to the inhibition of Ets-1 expression and TGF-β1/Smads signaling pathway.

Objective:To evaluate the efficacy of arthroscopic suture combined with perforator tendon double reduction and Endobutton plate technique for the treatment of tibial avulsion fracture of posterior cruciate ligament (PCL).Methods:A retrospective case series study was conducted to analyze the clinical data of 32 patients with tibial avulsion fracture of PCL, who were admitted to First People′s Hospital of Huizhou from January 2018 to October 2022. There were 19 males and 13 females, with age range of 18-65 years [(42.8±15.0)years]. By the Meyers-Mckeever classicfication, 27 patients were type II, and 5 type III. All the patients were treated with arthroscopic suture combined with perforator tendon double reduction and Endobutton plate technique to fix PCL tibial avulsion fracture at the insertion point. The fracture reduction was observed by X-ray films before discharge. Three months postoperatively, the fracture healing was observed by CT imaging, and the stability of knee joint was evaluated by posterior drawer test. The knee active range of motion, visual analogue score (VAS), Lysholm score, and International Knee Documentation Committee (IKDC) score were used to evaluate the pain and knee function before operation, at 3 months after operation, and at the last follow-up. Intraoperative and postoperative complications were observed.Results:All the patients were followed up for 6-36 months [(19.7±8.7)months]. The X-ray films showed satisfactory reduction of the fracture before discharge, and the CT showed that the fracture line was blurred to various degrees at 3 months after operation. Two patients had degree I positive posterior drawer test at 3 months after operation, and the rest were negative. The knee active range of motion was improved from 54.2°(45.0°, 70.0°) preoperatively to 124.8°(120.0°, 130.0°) at 3 months postoperatively and to 130.6°(125.0°, 135.0°) at the last follow-up (all P<0.01). The VAS was decreased from 5.1(3.0, 7.0)points preoperatively to 1.2(1.0, 2.0)points at 3 months postoperatively and to 0.1(0.0, 0.0)points at the last follow-up (all P<0.01). The Lysholm score was improved from 31.2(24.3, 37.0)points preoperatively to 73.6(69.3, 78.8)points at 3 months postoperatively and 92.6(91.0, 95.0)points at the last follow-up (all P<0.01). The IKDC score was increased from (20.9±7.1)points preoperatively to (59.2±8.9)points at 3 months postoperatively and to (77.5±7.0)points at the last follow-up (all P<0.01). Compared with 3 months after operation, the last follow-up showed significantly improved knee active range of motion, significantly decreased VAS, and significantly increased Lysholm score and IKDC score (all P<0.05). No intraoperative nerve vascular injury, postoperative incision infection, or lower limb venous thrombosis was observed. Conclusion:Arthroscopic suture combined with perforator tendon double reduction and Endobutton plate technique can achieve good reduction and high healing rate in patients with tibial avulsion fracture of PCL, with improved joint mobility, relieved pain, good recovery of knee joint function, and few complications.

BACKGROUND: Implantation of bone marrow mesenchymal stem cells (BMSCs) is a potential alternative for promoting bone defects healing or osseointegration in osteoporosis. However, the reactive oxygen species (ROS) accumulated and excessive inflammation in the osteoporotic microenvironment could weaken the self-replication and multi-directional differentiation of transplanted BMSCs. METHODS: In this study, to improve the hostile microenvironment in osteoporosis, Poloxamer 407 and hyaluronic acid (HA) was crosslinked to synthetize a thermos-responsive and injectable hydrogel to load MnO2 nanoparticles as a protective carrier (MnO2 @Pol/HA hydrogel) for delivering BMSCs. RESULTS: The resulting MnO2 @Pol/HA hydrogel processed excellent biocompatibility and durable retention time, and can eliminate accumulated ROS effectively, thereby protecting BMSCs from ROS-mediated inhibition of cell viability, including survival, proliferation, and osteogenic differentiation. In osteoporotic bone defects, implanting of this BMSCs incorporated MnO2 @Pol/HA hydrogel significantly eliminated ROS level in bone marrow and bone tissue, induced macrophages polarization from M1 to M2 phenotype, decreased the expression of pro-inflammatory cytokines (e.g., TNFa, IL-1b, and IL-6) and osteogenic related factors (e.g., TGF-b and PDGF). CONCLUSION This hydrogel-based BMSCs protected delivery strategy indicated better bone repair effect than BMSCs delivering or MnO2 @Pol/HA hydrogel implantation singly, which providing a potential alternative strategy for enhancing osteoporotic bone defects healing.

There remains a significant gap in our quantitative understanding of crosstalk between apoptosis and necroptosis pathways. By employing the SWATH-MS technique, we quantified absolute amounts of up to thousands of proteins in dynamic assembling/de-assembling of TNF signaling complexes. Combining SWATH-MS-based network modeling and experimental validation, we found that when RIP1 level is below ~1000 molecules/cell (mpc), the cell solely undergoes TRADD-dependent apoptosis. When RIP1 is above ~1000 mpc, pro-caspase-8 and RIP3 are recruited to necrosome respectively with linear and nonlinear dependence on RIP1 amount, which well explains the co-occurrence of apoptosis and necroptosis and the paradoxical observations that RIP1 is required for necroptosis but its increase down-regulates necroptosis. Higher amount of RIP1 (>~46,000 mpc) suppresses apoptosis, leading to necroptosis alone. The relation between RIP1 level and occurrence of necroptosis or total cell death is biphasic. Our study provides a resource for encoding the complexity of TNF signaling and a quantitative picture how distinct dynamic interplay among proteins function as basis sets in signaling complexes, enabling RIP1 to play diverse roles in governing cell fate decisions.
Animals ; Apoptosis ; Caspase 8/metabolism* ; GTPase-Activating Proteins/metabolism* ; HEK293 Cells ; Humans ; Mice ; Mice, Knockout ; Necroptosis ; Receptor-Interacting Protein Serine-Threonine Kinases/metabolism*

Objective:To explore the correlations of α-melanocyte stimulating hormone ( α-MSH) levels in serum and synovial fluid with progression of primary knee osteoarthritis (KOA). Methods:A retrospective analysis was conducted of the 96 patients who had been diagnosed as primary KOA at Department of Orthopedics, The First Hospital of Huizhou from October 2018 to October 2019. Radiographic severity of KOA was determined by Kellgren-Lawrence (K-L) grades; α-MSH levels were measured by enzyme-linked immunosorbent assay (ELISA). Levels of pro-inflammatory cytokine interleukin-1 β (IL-1 β) and matrix metalloproteinase-3 (MMP-3) were also detected. Another 64 patients with patellar dislocation, matched in age and gender, were enrolled as controls. The Numeric Pain Scale (NPS) and revised Oxford Knee Score (OKS) were employed to evaluate their symptomatic severity. Receiver operating characteristics (ROC) curve was used to compare α-MSH, IL-1 β and MMP-3 with regard to their diagnostic values in the K-L grading. Results:There were no statistically significant difference in age, gender and body mass index between the 2 groups, showing they were comparable ( P> 0.05). The α-MSH levels in synovial fluid were significantly lower in the KOA patients than in the controls [(16.9±3.8) pg/mL versus (18.8±2.7) pg/mL] ( P<0.001); there were no significant differences between the KOA patients and the controls in the serum α-MSH levels [(24.9±1.8) pg/mL versus (24.8±1.7) pg/mL] ( P>0.05). The α-MSH levels in synovial fluid were negatively correlated with K-L grades ( r=-0.382, P<0.001) and negatively correlated with NPS ( r=-0.382, P<0.001) but positively correlated with OKS ( r=0.339, P<0.001). Moreover, the α-MSH levels in synovial fluid were negatively correlated with the IL-1 β levels in synovial fluid ( r=-0.483, P<0.001) and with the MMP-3 levels in synovial fluid ( r=-0.336, P< 0.001). Conclusions:The level of serum α-MSH may not be correlated with the progression of KOA but the synovial fluid α-MSH is negatively correlated with the progression of KOA. Therefore, the expression level of α-MSH in joint synovial fluid can be used as a potential biomarker for assessment of severity of knee osteoarthritis.

Objective:To investigate the correlation of mRNA and protein levels of Ghrelin in shoulder tissue and synovial fluid with severity of rotator cuff tear(RCT) and frozen shoulder(FS).Methods:Recruited for this study were 66 RCT and 66 FS patients who had been definitely diagnosed at Department of Orthopedics, The First Hospital of Huizhou from January 2018 to September 2019. Another 60 patients with rotator instability were recruited as a control group. After the severity of RCT was evaluated by ultrasonographic images, the RCT group was further divided into 3 subgroups according to the severity of RCT: a massive full thickness tear (MFTT) subgroup, a non massive full thickness tear (MFTT) subgroup, and a partial thickness tear (PTT) subgroup. The FS group was conventionally divided into 31 cases of acute phase and 35 cases of adhesive phase. Samples of subacromial bursa and shoulder joint bursa tissues and shoulder joint synovial fluid were collected. The expression of Ghrelin mRNA was detected by RT-PCR and the expression of Ghrelin protein in joint synovial fluid was detected by ELISA. The symptoms and functions of shoulder joint were evaluated by visual analogue scale (VAS) and Constant-Murley functional score.Results:There were no significant differences in gender or age between the RCT, FS and control groups, showing compatibility ( P>0.05).The expression of Ghrelin mRNA in subacromial synovial capsule and shoulder joint capsule and the expression of Ghrelin protein in shoulder synovial fluid in the RCT and FS groups were significantly lower than those in the control group ( P<0.05).The expression of Ghrelin mRNA in subacromial synovial capsule and shoulder joint capsule and the expression of Ghrelin protein in shoulder synovial fluid in the PTT subgroup were the highest, followed sequentially by NMFTT and MFTT subgroups, with significant differences between subgroups ( P<0.05). For FS patients, the expression levels of Ghrelin mRNA and protein in the acute phase were significantly lower than in the adhesive phase ( P<0.05). The relative expression of Ghrelin mRNA in joint capsule and the expression of Ghrelin protein in shoulder synovial fluid were negatively correlated with VAS scores and IL-6 levels (all P<0.05), and positively correlated with the Constant-Murley scores (all P<0.05). Conclusions:The expression of Ghrelin in shoulder tissue and synovial fluid is negatively correlated with the progress of rotator cuff tear and frozen shoulder.Local supplementation of Ghrelin may be a potential therapy for rotator cuff tear and frozen shoulder.

OBJECTIVETo investigate the prevalence of serum antibodies against Japanese encephalitis virus (JEV) in bats.

METHODSBlood samples from the heart were obtained from bats captured in Guangdong and Hainan Provinces in 2013. The anti-JEV antibodies in bat sera were tested using indirect ELISA and virus neutralization test.

RESULTSA total of 201 bat serum samples were tested, in which the total positivity rate of anti-JEV antibodies was 46.27% (93/201). The positive rate of anti-JEV antibodies in bats from Hainan and Guangdong Provinces was 88.89% (48/54) and 30.61% (45/147), respectively. All the samples from Rousettus leschenaultia, Miniopterus schreibersii, Pipistrellus abramus, and Rhinolophus macrotis were positive for anti-JEV antibodies, and up to 95.56% (43/45) of the samples from Miniopterus schreibersii (from Hainan Province) yielded positive results. Of the 28 samples with positive results by indirect ELISA, 15 showed positive results in virus neutralization test (53.57%) with neutralization antibody titers ranging from 1:10 to 1:28.22.

CONCLUSIONBats from different regions and of different species can be naturally infected with JEV and have a high prevalence of anti-JEV antibodies in their sera. The role of bats in the natural cycle of JEV awaits further study.

Animals ; Antibodies, Viral ; blood ; China ; Chiroptera ; immunology ; virology ; Encephalitis Virus, Japanese ; Enzyme-Linked Immunosorbent Assay ; Neutralization Tests

Objective To investigate the prevalence of serum antibodies against Japanese encephalitis virus (JEV) in bats. Methods Blood samples from the heart were obtained from bats captured in Guangdong and Hainan Provinces in 2013. The anti-JEV antibodies in bat sera were tested using indirect ELISA and virus neutralization test. Results A total of 201 bat serum samples were tested, in which the total positivity rate of anti-JEV antibodies was 46.27%(93/201). The positive rate of anti-JEV antibodies in bats from Hainan and Guangdong Provinces was 88.89% (48/54) and 30.61% (45/147), respectively. All the samples from Rousettus leschenaultia, Miniopterus schreibersii, Pipistrellus abramus, and Rhinolophus macrotis were positive for anti-JEV antibodies, and up to 95.56%(43/45) of the samples from Miniopterus schreibersii (from Hainan Province) yielded positive results. Of the 28 samples with positive results by indirect ELISA, 15 showed positive results in virus neutralization test (53.57%) with neutralization antibody titers ranging from 1∶10 to 1∶28.22. Conclusion Bats from different regions and of different species can be naturally infected with JEV and have a high prevalence of anti-JEV antibodies in their sera. The role of bats in the natural cycle of JEV awaits further study.