Adjuvants are an important part of vaccines. In order to overcome the weaknesses of traditional adjuvants, it is essential to develop new adjuvants. At present, CpG ODN has been used as vaccine adjuvants in clinical and preclinical studies of preventive antiviral, anti-parasitic and therapeutic tumor vaccines. Hepatitis B vaccine and COVID-19 vaccine with CpG ODN 1018 as adjuvant have been approved for marketing. In addition to single application, it is one of the most important research directions to develop new delivery systems of anti-tumor vaccines(such as nanoparticles and liposomes)with CpG ODN as adjuvant, and the delivery efficiency and targeting of CpG ODN need to be improved. In addition to the traditional subcutaneous and intramuscular immunization routes, CpG ODN is also being applied to the development of intranasal mucosal immune adjuvants, and its efficacy and mechanism as mucosal immune adjuvants have not been fully clarified.CpG ODN is an agonist of Toll-like receptor 9(TLR9), which can activate natural immunity and further mediate adaptive immune response. Its characteristic is that it can cause strong Th1 immune response. Although many clinical studies showed that CpG ODN adjuvants only cause slight and short-term adverse reactions, some studies also showed that CpG ODN may cause or aggravate autoimmune diseases and inflammatory syndrome by promoting Th1 response. How to overcome this shortcoming of CpG ODN adjuvants may be a challenge for the development of CpG ODN adjuvants in the future. In this paper,the latest research progress and challenges of CpG ODN adjuvants are reviewed.

Objective:To analyze the perpendicular force on each tooth produced by the reverse-curve arch wires of various angles and depths using the method of 3-D finite element (3-D FEM) analysis. Methods:3-D FEM models of lower teeth, periodontal ligaments and alveolar bone as well as the 0.46 mm?0.64 mm inch stainless steel reverse-curve arch wires with different angles and depths were developed with the ANSYS finite element software, and the displacements of deformation of the reverse-curve arch wires were established on the models. Results:A ideal 3-D FEM was construct,including 2 709 elements and 1 969 nodes. The depth and angle of the reverse-curve arch wires produced obvious effect on the force exerted to teeth, and they could alter both the strength and the direction of the force. The force was mainly loaded on the canines, the second premolars and the molars, while less loaded on the incisors. Affected by the reverse-curve arch wires, the perpendicular force on the incisors were brought intrusively and on the molars were brought intrusively and distal upright; however, the canines and the bicuspids, in the perpendicular direction, moved along with the transformation of the angle and depth of the reverse-curve arch wires. When the angle of reverse-curve arch wires was fixed, the intrusive force on the incisors and molars was increased along with the increase of depth of the reverse-curve arch wires. Perpendicularly, the canines underwent a process from extrusion to intrusion, while the bicuspids were from intrusion to extrusion. When the depth of reverse-curve arch wires was fixed, the intrusive force on the incisors and molars was increased along with the increase of angle of the reverse-curve arch wires. The canines underwent a process from intrusion to extrusion.Conclusion:The variation of the angle and depth of the reverse-curve arch wires may produce evident effect on the force exerted to teeth.

Breast cancer is the principal cause of death in malig-nancy women , usually treated with the combination of surgery , chemotherapy , radiotherapy and endocrinotherapy .With the de-velopment of cell biology , molecular biology , immunology, im-munotherapy becomes a new field of breast cancer treatment .In this review, we discuss new findings in breast cancer immuno-therapy , including recent successes with bispecific antibodies and immune checkpoint blockade .We also discuss therapeutic cancer vaccines and highlight several additional immunotherapy modalities in early stages of development .

Objective To understand the plasma protein differential expression before and after cardiopulmonary bypass (CPB)through conducting the comparative proteomics study on rats in order to find the plasma markers with potential value in the early diagnosis of CPB resulted complications.Methods 10 adult male SD rats were divided into the experiment group and the con-trol group randomly (n=5),and took food and water freely before operation.The rat models of CPB were constructed in the experi-ment group.But no any CPB operation was administered in the control group in addition to anesthesia induction,arterial and venous puncture procedure.1 mL of blood sample was extracted for separating plasma before CPB and at the end of CPB in the two groups. The total plasma protein was purified.Then the 2-dimensional electrophoresis and the scanning imaging by ImageScanner were per-formed.The protein spots verified to be differential expression were performed the cutting,enzymolysis and peptide fragment ex-traction.Finally the mass spectrometer was adopted to conduct the analysis and identification.Results The number of visualized spots was increased significantly after CPB.17 protein spots with up-regulated expression were identified as differential expression caused by CPB.5 proteins were verified by mass spectrometer analysis and database research.They were gelsolin,haptoglobin,apo-lipoprotein A-1,immunoglobulin gamma-2b and Ba1-647 respectively.Conclusion CPB can cause the differential expression of plas-ma proteins in rat model.According to the function analysis,gelsolin,haptoglobin and apolipoprotein A-1 have the potentiality of be-ing the plasma markers for studying CPB complications.

Objective To clone and fuse the cDNA of human cytokeratin 9 in prokaryotic expression system,and purify and identify the fusion protein.Methods The cDNA fragment of human cytokeratin 9 was amplified from human keratinocyte (HaCaT) total RNA with specific primers.The PCR products were cloned into vector pET-28a,then the fusion protein of his-CK9 was induced by IPTG.The expressed fusion protein of his-CK9 was purified by nickel ion affinity chromatography and identified by SDS-PAGE and Western blot.Results The sequencing proved that the recombinant vector of the cDNA of CK9 was correct.The fusion protein of his-CK9 was induced to be expressed in E.coli.The fusion protein of his-CK9 was highly purified and his-CK9 showed specific binding to the commercialized antibodies of CK9.Conclusion The recombinant vector of pET-28a-CK9 has been successfully constructed,and the fusion protein of his-CK9 has been successfully expressed and purified.

OBJECTIVE:To investigate the therapeutic efficacy of edaravone combined with citicoline sodium on acute cere-bral infarction and its effects on the levels of oxidative stress and inflammatory factors. METHODS:108 patients with acute cere-bral infarction were randomly divided into edaravone group(single group)and edaravone+citicoline sodium group(drug combina-tion group),with 54 cases in each group. Based on routine treatment,single group was given Edaravone injection 30 mg added in-to 100 ml 0.9% Sodium chloride injection intravenously,bid,used up within 30 min each time;drug combination group was addi-tionally given Citicoline sodium injection 0.5 g added into 250 ml 0.9% Sodium chloride injection intravenously,qd,on the basis of single group. Treatment course of 2 groups lasted for 2 weeks. NIHSS,HDS,Barthel index,oxidant stress indicator and inflam-matory factors were compared between 2 groups before and after treatment. RESULTS:After treatment,the effective rate of NI-HSS in drug combination group was 81.48%,which was significantly higher than single group(53.70%),with statistical signifi-cance (χ2=9.511,P=0.002). HDS score and Barthel index of 2 groups were significantly increased after treatment,especially in drug combination group,with statistical significance(P<0.05). Compared with before treatment,contents of MDA and ET-1 in 2 groups were decreased significantly,while SOD activity and NO content were increased significantly;the inflammatory cytokines IL-6,IL-8,IL-12 and IL-16,TNF-α were all decreased gradually,with statistical significance (P<0.05);the improvement of each indicator in drug combination group was more significant than single group,with statistical significance(P<0.05). CONCLU-SIONS:Edaravone combined with citicoline sodium show good therapeutic efficacy in the treatment of acute cerebral infarction, can decrease the levels of oxidative stress and inflammation and promote the recovery of the neurological function and the daily liv-ing ability.

Objective To evaluate the role of neuron-restrictive silencer factor (NRSF) in the spinal cord in remifentanil-induced hyperalgesia in a mouse model of incisional pain (IP) .Methods Fifty-six male Kunming mice were randomly divided into 7 groups (n=8 each):control group (group C) ,IP group (group I) ,IP +remifentanil group (group IR ) , NRSF antisense oligonucleotide group (NAS group ) , IP + NRSF antisense oligonucleotide group (I+NAS group ) ,IP + remifentanil + NRSF mismatch oligonucleotide group (IR+NMS group) , and IP + remifentanil + NRSF antisense oligonucleotide group (IR + NAS group ) . Artificial cerebrospinal fluid 5 μl was injected intrathecally once a day for 3 consecutive days in C ,I and IR groups .NRSF antisense oligonucleotide NAS 10μg was injected intrathecally once a day for 3 consecutive days in NAS ,I+NAS and IR + NAS groups . NRSF mismatch oligonucleotide 10 μg was injected intrathecally once a day for 3 consecutive days in IR+NMS group .A 1-cm longitudinal incision was made through skin ,fascia and muscle of the plantar aspect of the right hindpaw to establish the model of incisional pain in sevoflurane-anesthetized rats .At 30 min after the last injection ,normal saline 0.4 ml was infused subcutaneously in C and NAS groups ,the model was established and normal saline 0.4 ml was subcutaneously infused simultaneously in I and I+NAS groups ,and the model was established and remifentanil 0.04 mg/kg was subcutaneously infused simultaneously in IR ,IR+NMS and IR+NAS groups .At 3 days before operation (T0 ) ,4 h before operation (T1 ) and 4 ,12 ,24 and 48 h after operation (T1-5 ) ,mechanical paw withdrawal threshold to von Frey stimuli (PMWT ) and paw withdrawal latency to thermal nociceptive stimulus (PTWL ) were measured .Results Compared with C group ,the PWMT and PWTL were significantly decreased at T2-5 in I ,IR ,I+NAS ,IR+NMS and IR+NAS groups ( P<0.05) ,and no significant change was found in the PWMT and PWTL at each time point in NAS group ( P>0.05 ) .Compared with I group ,the PWMT and PWTL were significantly decreased at T2-5 in IR and IR+NMS groups ( P<0.05) , and no significant change was found in the PWMT and PWTL at each time point in I +NAS group ( P>0.05) . Compared with IR group ,no significant change was found in the PWMT and PWTL at each time point in IR+NMS group ( P>0.05) ,and the PWMT and PWTL were significantly increased at T2-5 in IR+NAS group ( P<0.05) . Conclusion NRSF in the spinal cord is involved in the development and maintenance of hyperalgesia induced by remifentanil in a mouse model of IP .

To study the effect of batroxobin(DF-521) on atherosclerosis, we divided 50 Japanese big ear rabbits into control group and high-lipid group. After the atherosclerosis model was successfully established, the high-lipid rabbits were divided into 3 groups(placebo group, treatment group 1 and treatment group 2). Batroxobin was injected in the treatment groups, and saline was injected in placebo group and control group. Getting the aorta before, inter and after treatment, dyeing the lipid, endothelium, smooth muscle, collagen fibers of the vascular plaque(the elastic fibers are of autofluorescence), we observed them with the light microscope and laser scanning confocal microscope. From the results, we found that the atherosclerotic plaque in the treatment groups, tended to be static four weeks later, but there was no obvious difference between treatment group 1 and treatment group 2. These implied that batroxobin possessed the action of stabilizing the atherosclerotic plaque, but the dosage-effect was not clear and the principle needed more study.
Animals ; Arteriosclerosis ; drug therapy ; pathology ; Batroxobin ; therapeutic use ; Disease Models, Animal ; Female ; Fibrinolytic Agents ; therapeutic use ; Male ; Rabbits

OBJECTIVE: To determine the changes of Mu-opioid receptor (Mor) and neuron-restrictive silencer factor (NRSF) in periaquductal gray (PAG) in mouse models of remifentanil-induced postoperative hyperalgesia. METHODS: Thirty-two Kun-Ming mice were randomly divided into 4 groups (8 mice in each group): Group C (mice underwent a sham procedure and saline was infused subcutaneously over a period of 30 min), Group I (mice underwent a surgical incision and the same volume of saline), Group R (mice underwent a sham procedure and remifentanil was infused subcutaneously at the moment of surgical incision over a period of 30 min), and group IR (mice underwent a surgical incision and remifentanil). Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) tests were performed 24 h before the operation and 2, 6, 24, and 48 h after the operation. The specimens were collected after behavioral testings at 48 h. The expressions of Mor and NRSF in mice's PAG neurons were determined by Western blot. RESULTS: Mechanical allodynia and thermal hyperalgesia developed in Group I, R and IR (P<0.01). Intraoperative infusion of remifentanil enhanced mechanical allodynia and thermal hyperalgesia in mice with planta incision (P<0.01). In Group R and Group IR, the expression of Mor was significantly lower (P<0.01) and NRSF was significantly higher (P<0.01) when compared with Group C and Group I. CONCLUSION Intraoperative infusion of remifentanil induces postoperative hyperalgesia in mouse models, accompanied with decreased expressions of Mor and increased of NRSF level in PAG neurons, which may be involved in remifentanil induced hyperalgesia.
Animals ; Disease Models, Animal ; Hyperalgesia ; chemically induced ; Mice ; Pain, Postoperative ; Periaqueductal Gray ; drug effects ; metabolism ; Piperidines ; administration & dosage ; Receptors, Opioid, mu ; metabolism ; Remifentanil ; Repressor Proteins ; metabolism

Objective:A machine learning algorithm was used to develop a predictive model of self-injury among college students and to explore the high-risk factors for self-injury among college students.Methods:From November to December 2022, a convenience sample of 791 college students from a university in Hebei Province was selected.Whether the self-injurious behavior occurred or not was regarded as an outcome variable.The basic demographics data were collected for statistical analysis.The adolescent self-harm questionnaire, the acquired helplessness scale, the Chinese version of the interpersonal needs questionnaire, the adolescent life events scale, and the childhood traumatic experiences questionnaire were used for assessment.The predictor variables were statistically analyzed by SPSS 26.0 software, and the performance of the model was evaluated by random forest, support vector machine and logistic regression so as to predict the self-injury behavior of college students.The model performance was evaluated by the accuracy, F1 score, sensitivity, specificity, and AUC value of the model, and the optimal model was selected.Finally, the optimal model was used to analyze the high-risk factors of college students' self-injury behaviors.Results:(1) The results of one-way ANOVA showed that the detection rate of self-injury behavior among college students was 42.4%(335/791), and the detection rate of male students was significantly higher than that of female students ( χ2=14.139, P<0.05). Individuals with lower-middle monthly household income(RMB 3 000-5 999) had a significantly higher detection rate of self-injury behavior than those with other monthly household income( P<0.05). (2) The accuracy of random forest, support vector machine, and logistic regression models were 85.53%, 85.96%, and 68.86%, F1 scores were 0.853, 0.864, and 0.676, and sensitivities were 83.91%, 89.04%, and 64.91%, respectively.The AUCs of support vector machine, logistic regression models and random forest were 0.89, 0.73 and 0.92.(3) The top ten characteristic variables of high risk factors for college students' self-injury behaviors based on the random forest algorithm with better predictive efficacy were emotional abuse, frustration of belonging, helplessness, interpersonal relationship factor, despair, emotional neglect, academic stress factor, monthly family income, perception of tiredness, and health adaptation factor, in that order. Conclusions:Random forest is optimal for predicting self-injury behavior among college students compared to support vector machine and logistic regression.Factors influencing self-injury behavior among college students originate from environmental factors, individual factors and interpersonal factors.