Objective To explore the effect of δ-opioid receptor (DOR)on ischemic tolerance of rat brain.Methods The focal ischemic tolerance models of Sprague Dawley rats were established using the twice suture method with the middle cerebral artery occlusion (MCAO).A total of 24 male Sprague Dawley rats were randomly divided into sham group,ischemia (MCAO) group,sham +ischemia (sham+ MCAO) group and ischemic preconditioning + ischemia (IP+ MCAO) group (n=6,each).The neurological status was assessed using Zea-Longa neurological deficit scores at 7 days after cerebral infarction.The mRNA expressions of DOR,Bax,Bcl-2 in hippocampus in ischemic rat brain were detected by RT-PCR method.Results The Zea-Longa neurological deficit scores were 0.0±0.0,2.6±0.5,2.8±0.6 and 1.5±0.6 in Sham group,MCAO group,Sham+MCAO group and IP+ MCAO group,respectively at 7 days after cerebral infarction.The scores had significant differences among MCAO group,Sham+ MCAO group and IP+ MCAO group (both P＜0.01),but no difference between MCAO group and Sham+MCAO group(P＞0.05).The mRNA expressions of DOR and Bcl-2 were higher and Bax mRNA expression was lower in IP+MCAO group than in MCAO and Sham+ MCAO groups (all P＜0.01).Conclusions Ischemic preconditioning may increase the mRNA expressions of DOR and Bcl-2,reduce Bax mRNA expression,and improve the neurological status in rats.

BACKGROUND:Cerebral ischemia tolerance can promote proliferation of autologous neural stem cells in the hippocampus of cerebral infarction rats, but panaxtrial saponins effects on the proliferation of autologous neural stem cells in the brain have not been reported. OBJECTIVE:To explore the relationship of panaxtrial saponins, ischemic preconditioning and proliferation of endogenous neural stem cells in the hippocampus of rats at 7 days after cerebral infarction, and to observe the effect on neurobehavioral scores of rats after cerebral infarction. METHODS:Fifty Sprague-Dawley rats were included and randomly divided into five groups:sham group, ischemia group, ischemic control group, ischemic preconditioning group, and panaxtrial saponins group. In the latter four groups, acute models of cerebral infarction were established using Zea-Longa method. In the sham group, only an incision was made on the neck. The focal-focal ischemic tolerance models were established with twice suture method in the ischemic preconditioning and panaxtrial saponins groups. Sham operation was instead of ischemic preconditioning in the ischemic control group. In the panaxtrial saponins group, rats were given intraperitoneal injection of 100 mg/kg panaxtrial saponins at 7 days before modeling. RESULTS AND CONCLUSION:After 7 days of cerebral infarction, the neurobehavioral score and the number of neural stem cells in the hippocampus were significantly increased in the ischemia group (P<0.01);compared with the ischemia group, the neurobehavioral scores were lowered in the ischemic preconditioning and panaxtrial saponins groups (P<0.01), while the number of neural stem cells in the hippocampus was increased (P<0.01). However, there was no difference between the ischemic preconditioning and panaxtrial saponins groups (P>0.05). In addition, differences in the neurobehavioral scores and the number of neural stem cells in the hippocampus were insignificant between the ischemic control group and ischemia group (P>0.05). These findings indicate that panaxtrial saponins can play a role similar to ischemic tolerance, and thus improve neurologic impairment in rats with cerebral infarction.

Objective To investigate the effect of brain ischemic preconditioning (IP) combined with traditional Chinese medicine three seven three alcohol saponin (PTS) on proliferation of endogenous neural stem cells and the mRNA expressions of delta opioid receptor (DOR),Bax,Bcl-2 in hippocampus at 7d post middle cerebral artery occlusion (MCAO).Methods The focal-focal ischemic tolerance models were established with twice suture method.80 SD rats were included and randomly divided into 5 groups:sham group,MCAO group,sham+ MCAO group,IP+ MCAO group,PTS+MCAO group (n=16 each).We chose 10 SD rats from each group to evaluate their neurological status,and made BrdU fluorescent immunolabeling.In addition,we chose the other 6 SD rats to detect the expression levels of DOR,Bax and Bcl-2 mRNA in ischemic region in hippocampusby using RT-PCR.Animals were given one set of BrdU injections (on day 6,three times,4h apart,50mg/kg) to label the proliferating cells.The neurological status was assessed by using Zea Longa neurological deficit scores at 7 days following cerebral infarction.Results Zea longa neurologic deficit scores in MCAO group and sham+ MCAO) group had significantly differences with IP+ MCAO group and PTS+ MCAO group respectively at 7d post MCAO(P＜0.01).There was no significant differeuce in Zea-longa neurologic deficit scores between MCAO group versus sham+ MCAO group,and IP+ MCAO group versus PTS+ MCAO group(P＞0.05).The number of BrdU+ ceils in hippocampus had significant differences between IP+ MCAO and PTS+ MCAO groups at 7d post MCAOand three groups of sham,MCAO and sham+ MCAO respectively (P＜0.01).There was no difference in the number of BrdU+ cells between MCAO versus Sham + MCAO groups and IP + MCAO versus PTS+MCAO groups(P＞0.05).DOR and Bcl-2 mRNA expression levels were higher and Bax mRNA expression level was lower in IP+ MCAO group than in MCAO,Sham+ MCAO and PTS+MCAO groups (P＜0.01).There were no significant differences in DOR,Bcl-2 and Bax mRNA expressions among MCAO,Sham + MCAO and PTS + MCAO groups (P＞ 0.05).Conclusions Acute cerebral infarction can induce the proliferation of endogenous neural stem cells in hippocampus in SD rats.IPC can facilitate the proliferation of endogenous neural stem cells in hippocampus afteracute cerebral infarction,improve the symptoms of neurologic dysfunction,increase DOR and Bcl 2 mRNA expressions,and reduce Bax mRNA expression in SD rats.PTS can facilitate the proliferation of endogenous neural stem cells in hippocampus after acute cerebral infarction in SD rats,and improve the symptoms of neurologic dysfunction,but it has no influence on the expressions of DOR,Bcl-2 and Bax mRNA.

BACKGROUND:Proximal humerus internal locking system locking plate is the most commonly used fixation method for proximal humeral fractures,but its failure rate is still high in clinical practice.Reasonable screw placement is an important factor affecting the stability of internal fixation. OBJECTIVE:To investigate the distribution of humeral head screws in the treatment of proximal humeral fractures with proximal humerus internal locking system plate and its influence on internal fixation failure. METHODS:From January 2017 to December 2021,data from patients with proximal humeral fractures undergoing proximal humerus internal locking system plate in Third Affiliated Hospital of Guangzhou University of Chinese Medicine were retrospectively analyzed.A total of 124 patients were enrolled,including 16 males and 108 females,at the age of≥60 years.According to whether there was internal fixation failure after operation,they were divided into normal group(n=101)and internal fixation failure group(n=23).The patient's age,gender,fracture type,the integrity of the medial column,plate height,neck-shaft angle,whether the talus screw was inserted,and the number of humeral head screws,were collected.The humeral head was divided into eight zones according to the postoperative digital radiography anteroposterior film,and the distribution characteristics of the screws in each zone were collected,and the heat map of the screw distribution was drawn. RESULTS AND CONCLUSION:(1)There were no significant differences between the two groups in age,gender,fracture type,the integrity of the medial column,plate height,neck-shaft angle,whether to insert talus screws,and the number of humeral head screws(P>0.05).(2)The heat map showed that the humeral head screws were evenly distributed in the normal group,mainly scattered in zones 4,6,and 7.However,the screw distribution in the internal fixation failure group was not uniform,mainly concentrated in zones 4 and 6.In addition,in the ideal area of talus screws(7/8 zone),there were significantly more screws in the normal group than in the internal fixation failure group.(3)It is indicated that in the treatment of proximal humeral fractures with proximal humerus internal locking system plate,the uniform distribution of humeral head screws is the key to ensuring the internal fixation effect.A reasonable distribution of humeral head screws helps to improve the treatment effect and the success rate of internal fixation.

Objective:Anterior cruciate ligament injury is the most common type of knee joint ligament injury.Anterior cruciate ligament reconstruction has a high failure rate,with bone tunnel abnormalities as the most significant factor in these failures.Digital orthopedic technology can effectively develop implementation plans for the revision,thus increasing the success rate.This study aims to develop a surgical plan for anterior cruciate ligament revision by employing multiplanar reconstruction(MPR)for measuring bone tunnel position and diameter,and simulating bone tunnel creation via 3D printing preoperatively. Methods:A total of 12 patients who underwent anterior cruciate ligament revision at the Third Xiangya Hospital of Central South University between 2014 and 2021 were retrospectively studied.The data included patient demographics,preoperative formulated knee joint 3D printing models,and preoperative knee CT scans.The study measured the bone tunnel's diameter and position to guide the establishment of revision bone tunnels during surgery,reassessed the postoperative bone tunnels,and evaluated knee joint functional scores[including International Knee Documentation Committee Knee Evaluation Form(IKDC)score,Lysholm score,and Tegner exercise level score]. Results:Preoperative measurements revealed suboptimal femoral tunnels positions in 4 patients and tibial tunnels positions in 2 patients.MPR and 3D printing technology were used to guide the establishment of a new bone canal during surgery,and postoperative measurements were satisfactory for all patients.Preoperative measurements demonstrated the interclass correlation coefficient for femoral tunnels and tibial tunnels diameters were 0.843(P<0.05)and 0.889(P<0.001),respectively.Meanwhile,the intraclass correlation coefficient were 0.811(P<0.05)and 0.784(P<0.05),respectively.The intraoperative diameter of femoral and tibial tunnels showed excellent correlation with postoperative CT measurements,with intraclass correlation coefficient values of 0.995(P<0.001)and 0.987(P<0.001),respectively.All bone tunnel positions were within the normal range.At the final follow-up,knee joint function scores in all 12 patients improved significantly compared to pre-surgery(P<0.001),and the reoperation rate was zero. Conclusion:MPR and 3D printing technology can accurately measure the parameters of reconstructed anterior cruciate ligament bone tunnels.Personalized revision plans for patients with reconstruction failure enhances the success rate of revision surgery and improves patient prognosis.

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3-LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.