Opioid analgesics are currently known as the best analgesics. However， toxicity and side effects such as constipation， tolerance and addiction severely limit their clinical application. With the in-depth understanding of the signal transduction mechanism of opioid receptors and the continuous advancement of drug design technology， researchers have managed to develop many promising new methods to get low-toxic and more efficient opioid analgesics， which are different from the traditional morphine skeleton structure modifications. This article focuses on three new research strategies of G-protein biased activation，“ one drug-multiple targets” and peripheral activation. The basic principles of relative separation of analgesic activity and adverse drug reaction by each strategy are introduced， and the latest research progress of representative drugs is briefly reviewed. Among them， the recently approved novel opioid analgesics oliceridine and tegileridine are G-protein biased μ-opioid receptor agonists， Cebranopadol is a typical “one drug-multiple targets” analgesic， and NFEPP is a representative drug of peripheral opioid receptor agonists. The above several strategies complement each other and provide reference for the development of new opioid analgesic drugs.

OBJECTIVE To evaluate the cost-effectiveness of adebrelimab combined with chemotherapy versus chemotherapy alone in the first-line treatment of extensive stage small cell lung cancer from Chinese healthcare system perspective. METHODS Using the data obtained from the CAPSTONE-1 trial(230 cases for adebrelimab group, and 232 cases for chemotherapy group), Markov model was created for simulation of the disease development process of the extensive stage small cell lung cancer. The total costs, quality-adjusted life-years(QALYs) and incremental cost-effectiveness ratio(ICER) in each group were calculated. The sensitivity of key parameters was analyzed. RESULTS Compared with pure chemotherapy(etoposide plus carboplatin chemotherapy), the ICER of adebrelimab combined with chemotherapy was 157128.79 yuan·QALY−1 under the situation of charity assistance, and 351367.27 yuan·QALY −1 in the environment of no charity assistance. Sensitivity analysis showed that the utility and the cost of adebrelimab were the main influence parameter. CONCLUSION Adebrelimab combined with chemotherapy regimen has no cost-effective advantage versus chemotherapy alone in the treatment of extensive stage small cell lung cancer under the current economic level of China; the probability of adebrelimab combined with chemotherapy being cost- effectiveness was 44.5% under the situation of charity assistan.

Objective To preliminarily investigate the effects of estradiol on retinal microglia and retinal ganglion cells(RGCs)in rats with glucocorticoid-induced ocular hypertension(OHT).Methods Thirty-six male SD rats(36 eyes)were randomly divided into a control group,an OHT group,and an OHT estradiol-treated group(E2-OHT group),with 12 rats in each group.Among them,the rats in the OHT group and the E2-OHT group were given dexamethasone sodi-um phosphate injection under the conjunctiva,and the rats in the control group were injected with the same volume of ster-ile normal saline.Two weeks after modeling,the rats in the E2-OHT group were treated with estradiol eye drops in addition to subconjunctival injection of dexamethasone sodium phosphate.The eyeballs of all rats were removed 4 weeks after mod-eling.The changes in the number of RGCs and the activation of microglia were observed after immunofluorescence stai-ning,the expression levels of Brn3a and Iba1 proteins in the retina were detected by Western blot,and the relative expres-sion levels of tumor necrosis factor α(TNF-α)and interleukin 1 β(IL-1 β)mRNA were detected by real-time quantitative polymerase chain reaction.Results Among the three groups,the intraocular pressure(IOP)of rats showed no signifi-cant difference before modeling(all P＞0.05),but showed a significant difference at 1 week,2 weeks,3 weeks and 4 weeks after modeling(all P＜0.01).Compared with the control group,the IOP of rats in the OHT group at 1 week,2 weeks,3 weeks and 4 weeks after modeling increased significantly(all P＜0.01).Compared with the OHT group,the IOP of rats in the E2-OHT group showed no significant difference at 1 week and 2 weeks after modeling(both P＞0.05),but decreased significantly at 3 weeks and 4 weeks after modeling(both P＜0.01).The immunofluorescence staining results showed that the retinal microglia of rats in the control group were mainly concentrated in the inner plexiform layer,while the retinal microglia of rats in the OHT group migrated to the ganglion cell layer and had morphological changes(amoebic activation state).The morphology and distribution of rat retinal microglia in the E2-OHT group were basically the same as the retinal staining results of rats in the control group.Compared with the control group,the number of RGCs in the OHT group decreased,the relative expression levels of TNF-α and IL-1β mRNA and Iba1 protein increased,while the expression level of Brn3a protein decreased,and the differences were statistically significant(all P＜0.05).Compared with the OHT group,the rats in the E2-OHT group had an increased number of RGCs,a decreased relative expression level of TNF-α and IL-1 β mRNA and Ibal protein,and an increased expression level of Brn3a protein(all P＜0.05).Conclusion Estradiol can inhibit the activation of microglia,reduce the expression of TNF-α and IL-1β in the retina of rats with OHT,and reduce the damage to RGCs.

OBJECTIVE To investigate the effect and mechanism of action of KRT8 small interfering RNA(si-KRT8)in exosomes derived from patient serum with hypopharyngeal carcinoma on chemosensitivity to 5-fluorouracil(5-FU)in human pharyngeal squamous cell carcinoma FaDu cell line.METHODS The cancerous tissues of hypopharyngeal cancer patients and The serum,cancerous tissues and paracancerous tissues of drug-resistant patients after treatment were collected.A 5-FU-resistant cell line of FaDu(FaDu/R)was constructed for subsequent experiments.Exosomes were isolated from patient serum by ultrafast gradient centrifugation,identified using transmission electron microscopy and Western blot(WB)techniques.si-KRT8 was encapsulated into exosomes(Exosome@si-KRT8)using electroporation technology and subsequently used to treat cells.Real-time fluorescence quantitative PCR(RT-qPCR)and WB were used to detect the expression levels of KRT8 in different tissues,exosomes after electroporation of si-KRT8,and FaDu cells,respectively.Cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay,migration invasion technique and WB were used to detect the effects of Exosome@si-KRT8 on viability,apoptosis,and epithelial-mesenchymal transition of FaDu/R cells.RESULTS The expression level of KRT8 in drug-resistant tissues of hypopharyngeal carcinoma and FaDu/R cells was elevated compared with that in paraneoplastic tissues,cancer tissues and normal FaDu cells(t=15.79,P＜0.01).Isolated patient serum exosomes showed a double membrane structure and expressed both CD63 and TSG101 proteins,and KRT8 expression in exosomes was decreased after electro-transfection with si-KRT8(t=6.70,P＜0.01).Exosome@si-KRT8 inhibited KRT8 protein expression levels in FaDu/R cells(t=123.50,P＜0.01).Compared with the 5-FU group and the 5-FU+Exosome group,Exosome@si-KRT8 was able to inhibit the viability of FaDu/R cells(t=17.07,P＜0.01),promote the level of apoptosis in FaDu/R cells,and inhibit the expression of drug-resistance-associated proteins in FaDu/R cells(P-gp:t=103.20,MDR1:t=238.60,P＜0.01),and Exosome@si-KRT8 was able to suppress the expression of metastasis of FaDu/R cells(t=42.30,t=122.00,P＜0.01)and promoted the expression of E-cadherin while inhibiting the expression level of N-cadherin(t=130.80,t=83.90,P＜0.01).CONCLUSION Serum-derived exosome encapsulation of si-KRT8 enhances chemosensitivity of hypopharyngeal carcinoma cells and its mechanism of action may be related to inhibition of epithelial-mesenchymal transition.

Objective:To evaluate the effect of esketamine combined with ultrasound-guided dorsal penile nerve block (DPNB) on negative postoperative behavioral changes (NPOBCs) in pediatric patients undergoing circumcision under general anesthesia.Methods:One-hundred and ninety-five pediatric patients, aged 4-8 yr, with body mass index of 10-35 kg, of American Society of Anesthesiologists Physical Status classificationⅠ or Ⅱ, undergoing elective circumcision under general anesthesia, were selected and divided into 3 groups ( n=65 each) using a random number table method: esketamine group (group E), DPNB group (group D) and esketamine combined with DPNB group (group ED). Propofol 1.5 mg/kg was intravenously injected, and the patients were admitted to the operating room after consciousness disappeared in the 3 groups. Esketamine 0.5 mg/kg was intravenously injected in E and ED groups, and the equal volume of normal saline was given in group D. D and ED groups underwent bilateral DPNB with 0.25 % ropivacaine 0.15 ml/kg under ultrasound guidance, with the maximum total amount of the drug not exceeding 10 ml. Fentanyl 1.0 μg/kg and propofol 2.0 mg/kg were intravenously injected prior to the skin incision in the three groups. If intraoperative body movement occurred, propofol 10 mg was added, which could be repeated. The occurrence of intraoperative body movement, respiratory depression and amount of propofol added was recorded. When postoperative pain (FLACC score >4) occurred, flurbiprofen 1 mg/kg was intravenously injected for analgesia, and the usage of flurbiprofen was recorded. When emergence agitation(PEAD score>10) occurred, propofol 1 mg/kg was intravenously injected for sedation, and the occurrence of emergence agitation was recorded. Parents were followed up by telephone at 1, 7 and 30 days postoperatively to assess the occurrence of NPOBCs using the PHBQ scale. Results:Fifty-six patients in group E and 59 patients in D and ED groups finally completed the study.Compared with group E, the incidence of intraoperative body movement was significantly decreased, the amount of additional propofol was reduced, the emergence agitation score, incidence of emergence agitation and severe agitation and usage rate of postoperative flurbiprofen were decreased, and the incidence of separation anxiety at 7 and 30 days postoperatively was decreased in D and ED groups, and the incidence of intraoperative respiratory depression was significantly decreased, and the incidence of NPOBCs at 7 and 30 days postoperatively was decreased in group ED ( P<0.05). Compared with group D, the incidence of intraoperative respiratory depression was significantly decreased, the amount of additional propofol was decreased, the usage rate of postoperative flurbiprofen and incidence of sleep anxiety at 1 day postoperatively were decreased ( P<0.05), and no significant change was found in the incidence of NPOBCs at each time point after operation in group ED ( P>0.05). Conclusions:Esketamine combined with ultrasound-guided DPNB can reduce the occurrence of NPOBCs in pediatric patients undergoing circumcision under general anesthesia.

Objective:To analyze the characteristics of viral isolation and unique recombinant from untreated HIV-1 patients infected through sexual transmission and injection drug use, so as to provide evidence for understanding the biological characteristics and precise prevention and control of HIV-1 infection in different transmission routes.Methods:In view of the different HIV-1 transmission risks, newly diagnosed untreated HIV-1 patients from Beijing, Guangxi and Sichuan were carefully selected. Venous blood was collected to detect the viral load and CD4 + T cell count, and peripheral blood lymphocytes were isolated for virus isolation. Viral RNAs were extracted from the virus supernatant, and the near full-length genome sequences were obtained using in-house method, then the recombination patterns were determined. Results:Among the 65 HIV-1 infection, 32(49.2%), 20(30.8%) and 13(20.0%) were infected via men who have sex with men (MSM), heterosexual and injection drug use (IDU), respectively; genotypes mainly included 26(40.0%) CRF07_ BC, 23(35.4%) CRF01_ AE, and 9(13.8%) unique recombinant types (URFs). A total of 46 HIV-1 clinical strains were isolated. The positive rate of HIV-1 isolation was significantly negatively correlated with CD4 + T cells ( X2=4.22, P=0.04), but positively correlated with viral load ( X2=22.4, P<0.001); the multi-variate generalized estimating equations(GEE) model analysis of HIV-1 P24 antigen content showed similar result. In addition, GEE model showed a positive correlation between viral P24 antigen content and virus-producing culture time (52.14, 95% CI: 9.42~94.87, P=0.017). Viral growth curve analysis showed that the level of viral P24 antigen in MSM Group was significantly higher than that in heterosexual group and IDU group (adjusted P values were p<0.01 and P<0.05, respectively), on the 14th day after culture. The proportion of URFs in MSM Group was higher than that in heterosexual group, and the recombinant breakpoints in MSM Group were more than that in heterosexual group. Conclusions:MSM population was more sensitive to HIV-1 virus isolation; there was unique diversity of recombinant forms of HIV-1 among those with sexually transmitted infections, especially in the MSM population.

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.
Chromatography, Liquid ; Cloning, Molecular ; Escherichia coli ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; Tandem Mass Spectrometry

Objective To investigate the change rules of the lumbar vertebral bone marrow fat fraction(FF)and T2* values at different bone mineral density(BMD)groups,and their relevances and the applications on the diagnosis of the primary osteoporosis. Methods The patients who underwent the dual energy X-ray absorptiometry(DXA)and the routine lumbar MRI scan,mDIXON-Quant scan were collected,and the BMD,T-score,FF and T2* values of L1-L4 were measured.According to the T-score,the vertebral bodies were divided into the normal group,the osteopenia group and the osteoporosis group.The differences of the FF,T2* values and BMD between each group were analyzed by one-way analysis of variance,and their correlations with BMD were analyzed by Pearson correlation,the diagnostic effects of them were evaluated by receiver operating characteristic curve(ROC curve).Results The differences of the FF ,T2 * values and BMD among the three groups were statistically significant (P < 0 .05) .The correlations of the FF and T2 * values with BMD were negative (r = - 0 .628 ,P < 0 .05 and r = - 0 .468 ,P < 0 .05 ,respectively) .The area under curve (AUC) of the FF and T2 * values were 0 .82 ± 0 .03 ,0 .79 ± 0 .03 ,respectively .Conclusion mDIXON-Quant which can accuratly quantify fat content could evaluate the lumbar vertebral bone marrow fat content ,reflect the changes of the bone quality ,provide valuable information for the diagnosis of osteoporosis ,and is helpful to predict the risks of the lumbar fragility fracture.

Objective To identify the efficacy of granulocyte-macrophage colony-stimulating factor (GM-CSF) gargle in patients who had oral mucositis after allogeneic hematopoietic stem cell transplantation. Methods A total of 134 patients were enrolled in this study from 2014 to 2015. They were randomly divided into two groups:the experimental group (n=65) and the control group (n=69). Both groups received preventive measures for oral mucositis. But once oral mucositis occurred, the control group continued with the routine nursing measure, while the experimental group added GM-CSF gargle based on previous routine nursing measure. The effective rate and healing time were compared between two groups. Results The effective rate of the experimental group (81.54%, 53/65) was significantly higher than that of the control group (24.64%,17/69) (χ2=43.434, P=0.000). The median healing time in the experimental group was 4.5 days, shorter than 9.0 days in the control group (Z=-5.379, P< 0.01). Conclusions GM-CSF gargle can control the oral mucositis after allogeneic hematopoietic stem cell transplantation.

Neurogenic detrusor overactivity (NDO) often occurs after spinal cord injury, which often causes urinary tract infection, vesi-coureteral reflux, or even renal failure, and seriously impacts on the patient's quality of life. This paper reviewed the mechanism, the com-mon treatment methods, and neuromodulation theray of NDO after spinal cord injury, and elaborated percutaneous posterior tibial nerve stimulation, pudendal nerve regulation and the sacral neuromodulation respectively.