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 explore the function of risk management in outpatient blood collection work under the conditions of informationization. Method:This paper retrospectively reviewed the nursing risk management in out-patient blood collection work from January 2014 to January 2015 in our hospital, analyzed the causes of risk in out-patient blood collection work, evaluated the possible adverse outcomes, and put forward the measures to prevent and control risks. Results:Through the nursing risk management, the nurses′ risk prevention consciousness was enhanced, as well, both nurse and patient satisfaction was improved. Conclusion:Application of risk management in outpatient blood collection work could improve the quality of nursing, conform to the ethical requirements of guaranteeing patient safety, and effectively reduce the incidence of medical risks and accident.

Objective To compare the value of assessment with DWI and contrast-enhanced MRI (CE-MRI) in activity of sacroiliitis of patients with ankylosing spondylitis(AS).Methods Ninety-six patients conforming to modified New York criteria were prospectively collectedas the AS group, and twenty-one healthy volunteers were enrolled into the control group. According to the Bath AS disease activity index (BASDAI), erythrocyte sedimentation rate and C-reaction protein, AS patients were divided into the active AS group (n=60) and the chronic AS group (n=36) . All subjects were performed with conventional MRI, DWI and CE-MRI of bilateral sacroiliac joints. The MRI manifestations were reviewed and the ADC values and signal intensity enhancement rate (ΔSI) were measured.ANOVA was performed for the comparison ofΔSI and ADC values among active AS group, chronic AS group and control group with BASDAI and lab test results as the gold standards. ROC was analyzed with ΔSI and ADC values for activity of AS and paired
samples t test was obtained to comparethe areas under the ROC ofΔSI and ADC values.Results Among 96 cases of AS patients, MRI of sacroiliac jointsshowed that 62 cases had subchondral bone edema (57 cases of active group, 5 cases of chronic group), that 11 cases had bone surface erosion(4 cases of active group, 7 cases of chronic group), that 15 cases had bone sclerosis(6 cases of active group, 9 cases of chronic group) and that 58 cases had fat deposition on the sacroiliac joints (27 cases of active group, 31 cases of chronic group). The ΔSI values of the active group, the chronic group and control group were respectively (2.51 ± 1.69)%,(1.19 ± 0.67)%and(0.75 ± 0.21)%, and the ADCvalues were(1.33 ± 0.33)× 10-3,(1.00 ± 0.43)× 10-3 and(0.38±0.13)×10-3mm2/s. There were significant differences forΔSI and ADC values among three groups (F=18.375, 16.366. P<0.01), and statistical significance ofΔSI and ADC values were found between every two groups of three(P< 0.05).The area under the ROC between ΔSI and ADC to determine activity of AS patients were respectively 0.814 and 0.730, which had nostatistical significance(t=1.632, P=0.103). The sensitivity and specificity to determine activity of AS patients byΔSI=1.44%were 81.67%and 80.00%.The sensitivity and specificity to determine activity of AS patients by ADC=1.15 × 10-3/mm2 were 76.67% and 71.43%.Conclusion DWI and CE-MRI performed equally in detecting activity of AS patients.

BACKGROUND:Foreign injectable sulphate calcium has good biocompatibility, injectability and in situimmobilization, moulding based on adaptation to the shape of bone defects, but the price is expensive. OBJECTIVE:To explore the optimal fabricating parameters of bone repair materials with α-calciumsulfate hemihydrates as the main component, and to study the performance and characterization METHODS:α-Calciumsulfate hemihydrates powder was mixed with sodium hyaluronate at liquid-solid-ratios of 0.2, 0.25, 0.3, 0.35, 0.4 mL/g using vapor-heat method to prepare injectable bone materials. Performance, setting time and compressive strength of the injectable bone was detected. The best liquid-solid-ratio was 0.3 mL/g.α-Calcium sulfate hemihydrates powder was mixed with calcium sulfate dihydrate powder (1%, 2%, 3% mass fractionas) to fabricate injectable bone materials. Performance, setting time and compressive strength of the injectable bone was also detected; meanwhile, the biosafety of the injectable bone was determined. Theinjectable bone material that was made at the liquid-solid-ratio of 0.3 mL/g and by 2% calcium sulfate dihydrate was implanted into Ba-ma swine models of thoracic bone defects. At the time points of 8, 16 and 24 weeks after implantation, histological observation was done. RESULTS AND CONCLUSION: The injectable bone material was made at the liquid-solid-ratio of 0.3 mL/g and by 2% calcium sulfate dihydrate. The initial and final setting time was 4.0-5.0 minutes and 8.0-9.0 minutes, respectively. The compressive strength of the injectable bone reached (8.93±0.23) MPa. These findings indicate that the injectable boen material has good performance, initial setting time and compressive strength meeting the requirements of clinical application and good biosafety. Animal experiments show that the injectable bone can provide space for new bone in creeping substitution way by auto-degradation, with osteogenic activity.

Objective o evaluate the association between IL-28B ( rs12979860 ) polymorphism andantiviraltherapeutic effectbydetecting the genotype of interleukin-28B( IL-28 B) in patients with hepatitis C ( HCV ) .Methods Of total 1153 HCV patients, 303 diagnosed with CHC had been treated with pegylated interferon plus ribavirin for 24-48 weeks.IL-28B ( rs12979860 ) was genotyped by two-color fluorescent TaqMan assay.Results Among 1153 patients, CC, CT and TT genotype frequencies of IL-28B rs12979860 are 83.26%, 16.22%and 0.52%respectively.The results of HCV genotypingof 580 in 1153 cases, the frequencies of 1b, 2a and their non-1b/2a type are 63.45%, 35.00%and 1.55%respectively;In 303 CHC patients with clear medical history, the proportion of SVR was71.98% in patients with CC genotype and 16.90%in those with either the CT or TT genotypes.Logistic regression model was adopted to analyze the association of rs12979860 with SVR while adjusting for age, gender, viral load and HCV GT factors.Populations carrying combined genotype ( CT +TT) are making it harder to get SVR compared with those with CC genotype (OR, 95%CI:11.10,5.35-23.04;P<0.000 1).The percentages of SVR in HVC patients with 1b and 2a genotypeare 48.02% and 81.19% respectively.there is a statistically significant difference between these subgroups (χ2 =30.639,P<0.000 1).Conclusion IL-28B rs12979860 genotype is closely related to SVR in CHCpatients.Patients with CC genotype have a higher virus sustained response rate than those carrying CT or TT genotype.The SNP , rs12979860, might be applied as a predictor of clinical antiviral efficacy in the furture.

ObjectiveToinvestigateRUNX3genepromoter methylationincolorectal carcinogenesis and its prognostic significance. MethodsThe protein expression of RUNX3 was detected by immunohistochemistry and the methylation status of the RUNX3 was determined by methylation-specific PCR (MSP) in colorectal normal mucosa and cancer tissue from 65 patients,and adenoma from 28 patients.5-year overall survival rate was analysed according to RUNX3 methylation status from cancer patients.Kruskal-Wallis rank sum test,x2 or Fisher's exact test,Log-rank test and multivariable Cox regression analysis were used for statistical analysis.ResultsThe protein expression of RUNX3 gene in adenoma and cancer was 85％ (24/28) and 52％ (34/65),significantly lower than that in normal mucosa 94％ (61/65)( x2 =4.328,P =0.037 ; x2 =16.675,P =0.000),the difference between adenoma and cancer tissue was no statistic significance (x2 =3.266,P =0.071 ).No case showed RUNX3 methylation in normal mucosa,methylation rates in adenoma and cancer tissue were 21％ (6/28) and 35％ (23/65),significantly higher than in normal mucosa (P =0.000 ),but there was no statistic significance between adenoma and cancer tissue (x2 =1.766,P =0.183).The protein expression rate with RUNX3 methylation was 67％ (4/6) in adenoma,unmethylation 91％ (20/22) (P =0.191 ).The protein expression rate with RUNX3 methylation was 26％ (6/23) in cancer,unmethylation was 88％ (28/32).The presence of RUNX3 methylation was related to loss of protein ( x2 =9.810,P =0.002 ).5-year total survival rate with methylation in cancer was significantly lower with unmethylation ( x2 =5.87,P =0.016 ).Multivariate analysis showed RUNX3 methylayion wasanindependentprognosticfactoramongthefactorsanalyzed(P=0.033 ).ConclusionsRUNX3 methylation is important genetic event in colorectal carcinogenesis,possibly related to protein downregulation,and an independent prognostic factor for colorectal carcinoma.

Objective To investigate the acute effect of apolipoprotein E(apoE)on the intracellular free Ca~(2+) of rat cortical neurons. Methods The intracellular resting calcium level in cultured primary rat cortical neurons was measured by using confocal fluorescent imaging technique. MK-801, an N-methyl-D-aspartate (NMDA) receptor noncom-petitive antagonist, was employed to test potential function of apoE4 through blocking NMDN receptor. Results Acute application of apoE4, but not apoE3, significantly increased the resting [Ca~(2+)] i in a dose-and time-dependent manner (P <0. 01 or P <0. 05) , and MK-801 partly blocked the apoE4-induced elevation of resting [Ca~(2+)]i (P <0. 05 or P <0. 01). Conclusion Acute administration of ApoE4 disturbs calcium homeostasis, and the activation of NMDA receptor may play a critical role in the intracellular calcium overload induced by apoE4 and neurotoxicity.

Objective To examine whether microinjectlon of morphine into the rat thaiamle nucleus submedlus (Sin) could depress the bee venom (BV)-induced nociceptive behaviours. Methods In inflammatory pain model induced by BV subcutaneous injection into rat unilateral hind paw, the inhibitory effects of morphine microinjection into thalamic nucleus suhmedius (Sin) on the spontaneous nociecptlve behavior, heat hyperalgesia and tactile ailodynia, and the influence of naioxone on the morphine effects were observed in the rat. Results A single dose of morphine (5.0 μg, 0. 5μL) applied into the Sm ipsilaterni to the BV injected paw significantly depressed the spontaneous paw flinching response. Morphine also significantly increased the heat paw withdrawal iateneies in the bilateral hind paw and the tactile paw withdrawal threshold in the ipsilnteral hind paw 2 hours after BV injection. All these depressive effects could be effectively antagonized by pre-treatment with the opiuld receptor antagonist naloxone (1.0μg, 0. 5μL) in the Sm 5rain prior to morphine administration. Naloxone alone injected to the Sm had no effect on the BV-induecd nociceptive behavior. Conclusion These results suggest that Sm is involved in opioid receptor-mediated antt-nociception in the rat with the BV-induced inflammatory pain. Together with results from previous studies, it is likely that this effect is produced by activation of the Sm-ventrolateral orbital cortex-periaqueductal gray pathway, leading to activation of the brainstem descending inhibitory system and depression of the nodceptive inputs at the spinal cord level.

Objective To observe the changes in excitability of alpha-motoneuron responding to the muscle spindles afferent excitability in the model rats and explore whether the changes in this excitability to be changed as high frequency sinusoidal vibration acted on the soleus muscle.Methods The simulated weightlessness model was created by the tail-suspension.Mechanical vibrations at a frequency of 100 Hz with amplitude of 0.3 mm were acted on soleus muscle.The electromyogram of soleus muscle evoked by electrical stimulation of the sciatic nerve was recorded in control rats and HS rats.Results It was showed that HS and HFV did not influence the tendency of changes in amplitude of H reflex when the stimulation intensity increased.However,after 14-day HS,mean maximal motor response(Mmax)and mean maximal monosynaptic reflex/Mmax(Hmax/Mmax)both significantly decreased,respectively(P0.05).Conclusion It is suggested that HS should lead to the decrease in motoneurons excitability which is resulted from the reduction of muscle spindle afferent.HFV acted on soleus muscle of rats can improve the excitability of alpha-motoneurons in spinal cord during hindlimb suspension.

Objective To investigate the acute effect of apolipoprotein E(apoE)on the intracellular free Ca2+ of rat cortical neurons.Methods The intracellular resting calcium level in cultured primary rat cortical neurons was measured by using confocal fluorescent imaging technique. MK-801,an N-methyl-D-aspartate (NMDA) receptor noncompetitive antagonist,was employed to test potential function of apoE4 through blocking NMDN receptor. ResultsAcute application of apoE4,but not apoE3,significantly increased the resting [Ca2+]i in a dose-and time-dependent manner (P