Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To compare animal models of chronic heart failure(CHF)prepared by three different protocols,to establish a stable,reliable,and reproducible mouse model of CHF.Methods Twenty-five male C57BL/6J mice were divided randomly into four groups:a blank group,model A group(MA group),model B group(MB group),and model C group(MC group).The model groups adopted different preparation protocols for continuous injection of isoprenaline.The MA group and MB group were dose-decreasing models:MA group:subcutaneous injection of 10 mg/kg on day 1,5 mg/kg on day 2,2.5 mg/(kg·d)on days 3～30,total 30 days;and MB group:subcutaneous injection of 20 mg/kg on day 1,10 mg/kg on day 2,5 mg/(kg·d)on days 3～14,total 14 days.The MC group used a constant dose of intraperitoneal injection of 7.5 mg/(kg·d)for 28 days.The day after the final injection,the survival and model-formation rates for each group of mice were calculated.Cardiac function was measured by cardiac ultrasound and serum levels of N-terminal pro B-type natriuretic peptide,interleukin-6,and tumor necrosis factor-α were measured.Results CHF was successfully induced in all the model groups after all injections at the end of the fourth week.However,comprehensive test result showed that the MC model was the most stable.Conclusions An isoprenaline-induced mouse model of CHF using constant intraperitoneal injection of 7.5 mg/(kg·d)for 28 days may be the most suitable model for subsequent research on traditional Chinese medicine.

ObjectiveTo investigate the objective characteristics of tongue manifestations in patients with coronary heart disease （CHD）. MethodsA total of 315 participants with CHD were recruited in the CHD group， and 211 healthy participants who underwent physical examination were recruited as the healthy control group. In addition， according to the common comorbidities （primary hypertension， carotid atherosclerosis， type 2 diabetes mellitus， fatty liver， hyperlipidaemia， heart failure， and cerebral infarction） in 315 participants with CHD， each comorbidity was classified into a group of comorbidities with that disease and a group of non-comorbidities. Tongue images were captured using a TFDA-1 tongue diagnostic instrument to characterise the tongue body （TB） and tongue coating （TC）， comparing the RGB， HIS， and Lab colour spaces in the chromaticity index （R， red； G， green； B， blue； H， hue； I， intensity； S， saturation； L， lightness； a， red-green axis； b， yellow-blue axis）， the tongue coating thickness index （per-All）， contrast （CON）， angular second moment （ASM）， entropy （ENT）， and mean （MEAN） in texture metrics. ResultsCompared with the healthy control group， the characteristic indexes of tongue body in CHD group showed lower TB-R， TB-G， TB-B， TB-I， TB-L and higher TB-H， TB-b； and the characteristic indexes of tongue coating in CHD group showed lower TC-R， TC-B and higher TC-CON， TC-MEAN， TC-H， TC-b （P＜0.05 or P＜0.01）. Compared with non-combined primary hypertension group， CHD combined primary hypertension group showed higher per-All， TB-G， TB-L， and lower TB-a， TC-a （P＜0.05）； compared with the non-combined carotid atherosclerosis group， CHD combined carotid atherosclerosis group showed higher TB-CON， TB-ENT， TB-MEAN， and lower TB-ASM （P＜0.05 or P＜0.01）； compared with the non-combined type 2 diabetes mellitus group， CHD combined type 2 diabetes mellitus group showed lower per-All and higher TB-H （P＜0.05 or P＜0.01）； compared with the non-combined fatty liver group， CHD combined fatty liver group showed higher TB-CON， TB-MEAN， TB-ENT， and lower TB-ASM and TC-S （P＜0.05 or P＜0.01）； compared with the non-combined hyperlipidaemia group， CHD combined hyperlipidaemia group showed lower TB-S and TB-a （P＜0.05）； compared with non-combined heart failure group， CHD combined heart failure group showed lower TB-R， TB-G， TB-I， TB-L， and higher TB-a （P＜0.05 or P＜0.01）； compared with non-combined cerebral infarction group， CHD combined cerebral infarction group showed higher TC-CON， TC-ENT， TC-MEAN， and lower TC-ASM （P＜0.05 or P＜0.01）. ConclusionCompared to healthy individuals， patients with CHD tend to have darker tongue colours and rougher TC textures. Compared with non-comorbidity participants， those with primary hypertension tended to be lighter tongue colour and thicker tongue coating， those with carotid atherosclerosis had paler tongue body， those with type 2 diabetes mellitus had thinner tongue coating， those with fatty liver disease had paler tongue body and whiter tongue colour， those with hyperlipidaemia and heart failure had paler tongue colour， and those with cerebral infarction had rougher tongue texture.

Objective To establish a preliminary evaluation system for gastrointestinal qi stagnation syndrome.Methods On the basis of the systematic evaluation of medical literature in the early stage of the research group,24 high-frequency items were subjected to Delphi method,the item indexes were determined through three rounds of expert consultation,and the proportion value of the indexes was determined by AHP,and the evaluation system of gastrointestinal qi stagnation syndrome was initially constructed.Results A total of 84 valid questionnaires were collected by three rounds of Delphi method,including 15 in the first round,32 in the second round and 37 in the third round.According to the statistics,16 items including distention(stuffy)or distending pain or moving pain(epigastric,abdominal,etc.),belching,borborygmus,flatus,etc.were selected.The order of the proportion of the first level indexes obtained by the analytic hierarchy process from high to low is:clinical symptoms,pulse,tongue;The proportion of secondary indicators from high to low is as follows:distention(stuffy)or distending pain or moving pain(epigastric,abdominal,etc.),pulse string,greasy fur,thin white fur,slippery pulse string,pulse sinking string,light red tongue,flatus,borborygmus,belching,induced or aggravated in case of emotional distress,hiccup,abdominal mass,anorexia,vomiting,belching and swallowing acid.Conclusion Delphi method and analytic hierarchy process have been used to study gastrointestinal qi stagnation syndrome,and an evaluation system has been preliminarily formed.The index structure is reasonable,targeted and has strong clinical practicability.

Objective:To investigate the clinical efficacy of anterior approach combined with blocking plates and screws in the management of acetabular fracture involving the quadrilateral area.Methods:A retrospective case series analysis was performed for 16 patients with acetabular fracture involving the quadrilateral area admitted to First and Second Affiliated Hospital of Guangxi Medical University from January 2017 to January 2019. There were 12 males and 4 females，with the age of 21-66 years［（45.3±10.6）years］. According to Letournel-Judet classification，there were 9 patients with bi-column fracture，6 with anterior and posterior traverse fracture and 1 with anterior column fracture. A total of 9 patients were operated via the ilioinguinal approach and 7 via the lateral-rectus approach. Reduction and fixation of the pelvis and acetabulum were performed，using 3.5 mm cortical bone screws or plates to block the internal displacement of fracture in the quadrilateral body. The incision length，operation time and intraoperative blood loss were recorded. The quality of fracture reduction was assessed according to the Matta reduction criteria at postoperative 2 days and hip function by the modified Merle D'Aubigne-Postel score at postoperative 3 months and 12 months. Postoperative complications were observed.Results:All patients were followed up for 13-24 months［（16.1±2.9）months］. The ilioinguinal approach and lateral-rectus approach showed surgical incision of 12-26 cm［（18.6±4.0）cm］and 8-15 cm［（10.7±2.3）cm］，respectively. The operation time was 107-215 minutes［（159.2±27.8）minutes］and the intraoperative blood loss was 200-2，300 ml［（853.1±489.7）ml］. According to Matta reduction criteria，the results were excellent in 9 patients and good in 7. Three months after operation，the modified Merle D'Aubigne-Postel score was 11-18 points［（15.2±2.2）points］，which showed the results were excellent in 4 patients，good in 7，fair in 4 and poor in 1，with the excellent and good rate of 69%. Twelve months after operation，the modified Merle D'Aubigne-Postel score was 13-18 points［（16.9±1.4）points］，which showed the results were excellent in 7 patients，good in 8 and fair in 1，with the excellent and good rate of 94%. The liquefaction of post-surgical incision was seen in a patient，bladder injury in a patient，lateral femoral cutaneous nerve injury in a patient，and heterotopic ossification in a patient. There was no loosening or breakage of the internal fixation.Conclusion:For acetabular fracture involving the quadrilateral area，anterior approach combined with blocking plates and screws can prevent the displacement of quadrilateral fracture and attain satisfactory reductiongood hip function recovery and few complications.

Objective To investigate the effect of platelet-rich plasma(PRP) on starting of AnxA1 gene and PPARγ gene of adipose-derived stem cells (ADSCs) in rabbit.Methods Epididymal adipose tissue stem cells from New Zealand white rabbits,and the cells identified by morphology and inducing differentiation,the cells were cultured to the fourth generation,PRP and PPP (platelet-poor plasma) were prepared by traditional centrifugal method from abdominal aortic of rabbit;ADSCs were cultured in culture medium containing PRP (experimental group),PPP (control group) and all medium (blank group) for each 5％ for 24 h,48h and 72 h.Cells of each group were dissociated and total RNA extracted.AnxA1 gene and PPARγ gene were detected by RT-PCR.Results Primary ADSCs of rabbit grew in the way of long spindle swirly.The results of oil red O and alizarin red staining of the ADSCs were positive.AnxA1 gene and PPARγ gene of experimental group significantly increased from the result of RT-PCR (P＜0.05).Conclusions PRP can promote proliferation of the ADSCs of rabbit and increase the expression of AnxA1 gene and PPARγ gene significantly.

Objective To explore the optimal time for starting to drink plenty of water after 131I treatment in differentiated thyroid carcinoma(DTC) patients.Methods Totally 83 cases of DTC patients were randomly divided into three groups,and started to drink plenty of water at 12 h(group A),24 h(group B),and 36 h(group C) after treatment with 131I therapy.We measured and compared equivalent dose rate using the Inspector Alert gamma ray monitor at 1 meter in front of the patient's abdomen and neck at 6 h,12 h,24 h,36 h,48 h,72 h after taking 131I.We compared the discharge rate at different time and evaluated the curative effect.Results Equivalent dose rate of the abdomen at 24 h and 36 h after treatment and the discharge rate at 36 h and 48 h after treatment among three groups showed significant differences(P＜0.05).There were no statistically significant differences in the curative effect(P＞0.05).Conclusion Starting to drink plenty of water at 12 h after taking 131I can accelerate the decreasing of equivalent dose rate with no influence on the curative effect and improve the discharge rate at early.

Objective To explore the relationship between differentially expressed miR-19b-3p and cognitive function in patients with Alzheimer's disease (AD).Methods The miRNA expression profiles of patients with AD(AD group,n=30) or healthy elderly people (NC group,n=30) were analyzed by Illumina/Solexa high-throughput sequencing technique.The miRNA lentiviral plasmids were constructed and injected into the model of AD rats.The cognitive function of rats was analyzed with water maze test.The mimics and inhibitors were synthesized and transfected into SH-SY5Y cell lines.The protein expression of β-amyloid precursor protein cleavage enzyme 1 (BACE1) was detected by western blot.Results Compared with the NC group,the expression of miR-146a-5p (log2AD/Control =2.3),miR-195-5p (log2 AD/Control =10.2),miR-20b-5p(log2AD/Control =15.1) in serum of AD group was up-regulated and the expression of miR-19b-3p (log2 AD/Control =-8.0) and miR-125b-3p (log2 AD/Control =-15.3) was down-regulated (P＜ 0.05).Compared with the rats in the AD group((26.50±3.12) s),the rats in the AD+miR-19b-3p group ((15.33±2.78) s) showed significantly shorter escape latencies(P＜0.05).Compared with the NC group,the protein expression of BACE1 in miR-19b-3p mimetic group was decreased and the protein expression of BACE1 in inhibitor group was increased.Conclusion miR-19b-3p may improve the cognitive function of AD patients via regulating the expression of BACE1.

BACKGROUND:It has been an urgent problem of how to promote cartilage repair of the knee and shorten the total course through a tissue engineering approach. Fortunately, microfracture plus stem cel transplantation may open up a new path for this issue. OBJECTIVE:To investigate the clinic feasibility of arthroscopic microfracture technique plus stem cel transplantation for repair of articular cartilage injury of the knee. METHODS:From October 2010 to March 2012, a total of 16 patients with articular cartilage injury of the knee were enrol ed, including 12 males and 4 females, with the average age of 38.6 years (16-52). Al cases of cartilage injury were confirmed by arthroscopy. Autologous bone marrow was extracted from patients at 2 weeks before treatment to isolate, culture and amplify bone marrow mesenchymal stem cel s in vitro. The cel culture solution of 3-5 mL (about 107 cel s) was harvested. The articular cavity was clean by arthroscopy and microfracture technique was performed at the area of cartilage injury that was then covered with hemostatic gauze through a minimal y invasive incision and the prepared bone marrow mesenchymal stem cel s were injected. The knee was bandaged with the elastic bandage after aspirating the joint cavity effusion by vacuum suction. Functional exercises were performed early by CPM.RESULTS AND CONCLUSION:After fol ow-up of 4-18 months, there were 13 cases of excel ent, 2 cases of valid and 1 case of ineffective. According to Lysholm knee scores, the average scores were improved from 42 points (33-67 points) to 89 points (75-99 points) at 4 weeks after treatment. The function was satisfied and al patients were fol owed up without recurrence or worse. Under the arthroscopy, the combination of microfracture technique and autologous bone marrow mesenchymal stem cel transplantation is proved to be effective for articular cartilage injury of the knee and it can notably improve the clinic symptoms and recover the function of the knee.

BACKGROUND:Ligamentum flavum hypertrophy is one of the most important factors of lumbar spinal stenosis, but the molecular mechanism is stil not very clear. OBJECTIVE:To explore the role of basic fibroblast growth factor, connective tissue growth factor and transforming growth factor β1 in hypertrophy of the lumbar ligamentum flavum. METHODS: The ligamentum flavum samples were divided into three groups according to different diseases: control group (acquired from the patients with lumbar spinal canal tumor,n=6), lumbar disc herniation (LDH) group (acquired from the patients with LDH,n=6) and lumbar spinal stenosis (LSS) group (acquired from the patients with LSS,n=6). Then the mRNA expressions of basic fibroblast growth factor, connective tissue growth factor, transforming growth factor β1 and colagen I, III, V of the ligamentum flavum were detected using real-time quantitative RT-PCR method. The roles of basic fibroblast growth factor, connective tissue growth factor and transforming growth factor β1 were explored. RESULTS AND CONCLUSION:The expression of basic fibroblast growth factor mRNA in the LSS group was significantly higher than that in the LDH and control groups (bothP < 0.05); the expression of connective tissue growth factor mRNA was not found statisticaly different among the three groups, although it was slightly higher in the LSS group (P> 0.05); the expression of transforming growth factor β1 mRNA was significantly higher in the LSS group than in the LDH and control groups (bothP < 0.01). The colagen I mRNA expressed significantly higher in the LSS group than the LDH and control groups (bothP < 0.05), but both the colagen III and V mRNA showed no significant difference among the three groups (P> 0.05). This study indicate that both basic fibroblast growth factor and transforming growth factor β1 play important roles in the formation process of the lumbar ligamentum flavum hypertrophy, and the main type of the colagen in the hypertrophied ligamentum flavum is colagen I.