African swine fever virus(ASFV)I177L gene deletion vaccine is one of the key directions of African swine fever(ASF)live attenuated vaccine research and development.In order to effec-tively distinguish between the wild-type ASFV strain and the I177L gene-deleted strain,specific primers and probes were designed based on ASFV B646L and I177L genes,respectively.After screening and optimization,a duplex real-time PCR method was developed that can simultaneously detect these two genes.The results showed that ASFV B646L and I177L genes were detected spe-cifically and simultaneously by the method developed without cross-reactions with porcine circovir-us type 2,Seneca virus A,classical swine fever virus,foot-and-mouth disease virus,porcine respira-tory and reproductive syndrome virus.The detection limits of the duplex real-time PCR for recom-binant plasmids pUC57-B646L and pUC57-I177L were 1×103 copies/mL.The intra-and inter-as-say coefficients of variation were less than 4％,respectively.Detection of 122 pork and pork prod-ucts using the duplex real-time PCR developed and the real-time PCR recommended by WOAH showed that the coincidence rates of the two methods for B646L gene detection was 100％with two amplification curves appeared in the positive results of the established methods.The method established in this study can be used for the detection of ASFV I177L gene deletion strains,which provides technical support for ASF surveillance and epidemiological investigation.

OBJECTIVE To explore the material basis and potential mechanism of Kazakh classic prescription Wuzdekh （WZDK） in the treatment of enteritis. METHODS LC-MS/MS technology was used to analyze the chemical components in WZDK. Through network pharmacology and molecular docking technology， the main chemical components of WZDK were screened and the target was predicted； therapeutic effect and target of WZDK on acute enteritis were verified through in vivo experiments. The acute enteritis model of mice was induced by dextran sulfate sodium salt； the general condition of the mice was observed during administration and the disease activity index （DAI） score was calculated； pathological changes of the intestine and mRNA expression of core target were validated by HE staining and quantitative real-time PCR. RESULTS A total of 316 chemical components were obtained by LC-MS/MS. The core targets of network pharmacological analysis mainly included interleukin 1β（IL- 1β）， protein kinase B1 （AKT1）， tumor protein p53 （TP53）， IL-6， tumor necrosis factor （TNF） and so on. The results of molecular docking showed that chemical components such as mairin， lappadilactone， costunolide and dehydrocostus lactone were stable in binding to the core target. The results of in vivo experiment showed that， compared with model group， high dose （5.00 g/kg） of WZDK could significantly reduce the DAI score （P＜0.05）， improve inflammatory cell infiltration and mucosal tissue damage of colon tissue， and significantly down-regulated mRNA expressions of IL-6， TNF-α， IL-1β and TP53 in colon tissue（P＜ 0.05 or P＜0.01）. CONCLUSIONS Chemical components of WZDK such as mairin， lappadilactone， costunolide and dehydrocostus lactone may play the role of improving the imbalance of local inflammatory factors in the intestine and repairing damage of colonic mucosal tissue by down-regulating mRNA expressions of TNF-α， IL-6， IL-1β and TP53 in colon tissue.

Nitrate is the main form of inorganic nitrogen that crop absorbs, and nitrate transporter 2 (NRT2) is a high affinity transporter using nitrate as a specific substrate. When the available nitrate is limited, the high affinity transport systems are activated and play an important role in the process of nitrate absorption and transport. Most NRT2 cannot transport nitrates alone and require the assistance of a helper protein belonging to nitrate assimilation related family (NAR2) to complete the absorption or transport of nitrates. Crop nitrogen utilization efficiency is affected by environmental conditions, and there are differences between varieties, so it is of great significance to develop varieties with high nitrogen utilization efficiency. Sorghum bicolor has high stress tolerance and is more efficient in soil nitrogen uptake and utilization. The S. bicolor genome database was scanned to systematically analyze the gene structure, chromosomal localization, physicochemical properties, secondary structure and transmembrane domain, signal peptide and subcellular localization, promoter region cis-acting elements, phylogenetic evolution, single nucleotide polymorphism (SNP) recognition and annotation, and selection pressure of the gene family members. Through bioinformatics analysis, 5 NRT2 gene members (designated as SbNRT2-1a, SbNRT2-1b, SbNRT2-2, SbNRT2-3, and SbNRT2-4) and 2 NAR2 gene members (designated as SbNRT3-1 and SbNRT3-2) were identified, the number of which was less than that of foxtail millet. SbNRT2/3 were distributed on 3 chromosomes, and could be divided into four subfamilies. The genetic structure of the same subfamilies was highly similar. The average value of SbNRT2/3 hydrophilicity was positive, indicating that they were all hydrophobic proteins, whereas α-helix and random coil accounted for more than 70% of the total secondary structure. Subcellular localization occurred on plasma membrane, where SbNRT2 proteins did not contain signal peptides, but SbNRT3 proteins contained signal peptides. Further analysis revealed that the number of transmembrane domains of the SbNRT2s family members was greater than 10, while that of the SbNRT3s were 2. There was a close collinearity between NRT2/3s of S. bicolor and Zea mays. Protein domains analysis showed the presence of MFS_1 and NAR2 protein domains, which supported executing high affinity nitrate transport. Phylogenetic tree analysis showed that SbNRT2/3 were more closely related to those of Z. mays and Setaria italic. Analysis of gene promoter cis-acting elements indicated that the promoter region of SbNRT2/3 had several plant hormones and stress response elements, which might respond to growth and environmental cues. Gene expression heat map showed that SbNRT2-3 and SbNRT3-1 were induced by nitrate in the root and stem, respectively, and SbNRT2-4 and SbNRT2-3 were induced by low nitrogen in the root and stem. Non-synonymous SNP variants were found in SbNRT2-4 and SbNRT2-1a. Selection pressure analysis showed that the SbNRT2/3 were subject to purification and selection during evolution. The expression of SbNRT2/3 gene and the effect of aphid infection were consistent with the expression analysis results of genes in different tissues, and SbNRT2-1b and SbNRT3-1 were significantly expressed in the roots of aphid lines 5-27sug, and the expression levels of SbNRT2-3, SbNRT2-4 and SbNRT3-2 were significantly reduced in sorghum aphid infested leaves. Overall, genome-wide identification, expression and DNA variation analysis of NRT2/3 gene family of Sorghum bicolor provided a basis for elucidating the high efficiency of sorghum in nitrogen utilization.
Nitrate Transporters ; Nitrates/metabolism* ; Sorghum/metabolism* ; Anion Transport Proteins/metabolism* ; Phylogeny ; Protein Sorting Signals/genetics* ; Nitrogen/metabolism* ; DNA ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

OBJECTIVE To discuss the application effect of health care failure mode and effect analysis(HFMEA) and root cause analysis(RCA) in the intelligent management of narcotic use in the hospital. METHODS Set up an analysis team to use HFMEA to list potential failure modes in the flow chart of intelligent management of narcotic use in the hospital, and analyze and evaluate the key failure modes in combination with RCA, determine the root cause, and then formulate improvement measures and evaluate the effects after implementation. RESULTS After the implementation of the improvement measures, the risk priority number of the eight potential failure modes decreased significantly(P<0.05). The satisfaction of pharmacists, doctors and nurses with the intelligent management process of narcotic use in the hospital increased from (88.39±2.21)% to (98.04±0.51)%, from (87.79±1.36)% to (97.55±1.24)%, and from (90.79±1.39)% to (95.68±1.30)%, respectively. CONCLUSION HFMEA combined with RCA can reduce the risk of narcotic drug abuse, ensure drug quality and safety, reduce the occurrence of dispensing errors, and ensure the rational clinical use of narcotic drugs.

OBJECTIVE To review the application and development of automatic intelligent drug cabinet in hospitals at home and abroad. METHODS The relevant research articles published from 2010 to 2022 were retrieved from CNKI, Wanfang, VIP, PubMed and other databases. RESULTS The application of automatic intelligent drug cabinet could shorten the execution time of inpatient medical orders, improve the work efficiency of pharmacists and doctors and nurses, improve the quality of drug management in wards and promote the transformation of hospital pharmaceutical care. CONCLUSION The construction of decentralized dispensing mode with automated intelligent drug cabinets as the core has important theoretical guiding significance and broad application prospects for the automation, informatization, intelligence and whole process management of drugs in medical institutions, and also helps to improve the level of medical pharmacy service management.

The clinical data, laboratory test, and gene mutations were collected from a family with Liddle syndrome. Literatures on Liddle syndrome published in domestic and abroad since 1994 were reviewed and the types of gene mutations were summarized. The proband was diagnosed with hypertension at the age of 24. Laboratory test showed that serum potassium was 3.65 mmol/L, plasma renin was <0.5 mU/L, and plasma aldosterone was 1.5 ng/dL. Proband′s father was diagnosed with hypertension at the age of 34 with the serum potassium 3.34 mmol/L, plasma renin 3.72 mU/L, and plasma aldosterone 6.04 ng/dL. A nonsense mutation(1724G>A, p.Trp575*) in exon 13 of SCNN1G gene was detected in the proband and his father. In 288 cases from 107 families reported in the review of domestic and foreign literature, the incidence of hypertension, hypokalemia, and low renin/low aldosterone were 95.1%, 55.2%, and 49.6%, respectively. This case suggests that the clinical phenotype of Liddle syndrome is heterogeneous. Patients with early-onset hypertension, regardless of whether they are accompanied by hypokalemia, should be screened for renin-angiotensin-aldosterone and genetic testing related to Liddle syndrome should be further detected in patients with low plasma renin/aldosterone.

Objective:To investigate the treatment options for multiple myeloma patients with central nervous system involvement (CNS-MM) , as well as their clinical characteristics and prognostic factors.Methods:Between January 2011 and January 2022 our center diagnosed 18 people with CNS-MM. A retrospective analysis was done on the clinical information from the initial diagnosis and central nervous system involvement, and it was compared to 1∶3 matched newly diagnosed MM from the same period. Analysis was done on the clinical characteristics and survival rates of the two groups.Results:In patients with CNS-MM, the median time of onset was 14.2 (0.9-79.6) months and the median overall survival (OS) was 30.5 months from initial diagnosis and only 3.8 months in patients after CNS involvement. The CNS-MM patients showed more IgD type ( P=0.010) , severer anemia ( P=0.014) , a higher proportion of bone marrow plasma cells ( P=0.013) , more extramedullary lesions ( P=0.001) , and increased lactic dehydrogenase (LDH) ( P=0.009) when compared to the control group. Lenalidomide or pomalidomide-based combinations had higher rates of hematology and CNS remission than bortezomib or daratumumab-based regimens (75.0% vs 16.7% , P=0.019) . Patients who received IMiD-based regimens and had 2 high-risk factors at initial diagnosis (high LDH and extramedullary lesions) had a significantly lower incidence of CNS-MM ( P=0.026) . At the initial diagnosis, LDH ( P=0.008, HR=7.319, 95% CI 1.663-32.219) and extramedullary lesions ( P=0.006, HR=8.054, 95% CI 1.828-35.486) were independent risk factors for the occurrence of CNS-MM. Conclusion:Patients with CNS-MM had a poor prognosis. Patients with high LDH or extramedullary lesions at the time of the initial diagnosis are more likely to have CNS-MM. The prognosis of this patient may be improved by immunoregulator-based therapy.

Objective:To investigate the effects of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in treatment of high-risk multiple myeloma (MM) patients and its influencing factors of the prognosis.Methods:The clinical data of 44 high-risk MM patients treated with allo-HSCT in Changzheng Hospital Affiliated of Naval Military Medical University from April 2003 to March 2017 were retrospectively analyzed. The overall response rate (ORR), relapse rate, non-relapse-related death (NRM) rate, graft-versus-host disease (GVHD) incidence of patients were also analyzed. Kaplan-Meier was used to analyze the overall survival (OS) rate and progression-free survival (PFS) rate after transplantation. Cox proportional hazard model was used to make regression analysis of the factors affecting the prognosis.Results:Among 44 patients, 38 cases could be evaluated for efficacy after transplantation. The median follow-up time was 111 months (0-216) months, 22 cases survived, 22 cases died, 21 cases relapsed. Before transplantation, complete remission (CR) rate was 29.5%(13/44), very good partial remission(VGPR) rate was 45.5%(20/44), partial remission (PR) rate was 22.7% (10/44), stable disease (SD) rate was 2.3% (1/44); After transplantation, CR rate was 71.7%(27/38), VGPR rate was 13.2% (5/38), PR rate was 13.2% (5/38), the progression of the disease (PD) rate 2.6% (1/38). The 5-year OS rate and PFS rate was 51.8% and 47.8%, the 10-year OS rate and PFS rate was 51.3% and 43.1%, respectively; the 5-year and 10-year cumulative disease relapse rate was 38.6% and 45.4%, the 5-year cumulative NRM rate was 25.0%. Acute GVHD rate was 38.6% (17/44) and grade 3-4 acute GVHD rate was 6.8% (3/44); chronic GVHD rate was 27.3% (12/44). Cox univariate and multivariate analysis showed that the use of bortezomib before transplantation ( HR = 3.461, 95% CI 1.211-9.880, P = 0.020) and post-transplant infection ( HR = 0.283, 95% CI 0.098-0.819, P = 0.020) were independent factors affecting OS after transplantation. Conclusions:Allo-HSCT can overcome the high-risk factors of MM and is worth to try for high-risk MM patients. The use of bortezomib before transplantation and post-transplant infection can be important factors affecting OS after transplantation.

Chimeric antigen receptor T cell (CAR-T) is a kind of adoptive cell immunotherapy, in which T cells are genetically modified to exert targeted killing effect on tumors. CAR-T cell therapy has shown remarkable antitumor efficacy for the treatment of tumors, especially for hematological malignancies, but is less effective in solid tumors. Single-target CAR-T is prone to off-target effect during application, and there is a risk of relapse or more refractory treatment. The development of double-target or multi-target CAR-T is expected to extend the antigen coverage of target cells, effectively avoids antigen escape and prevents tumor recurrence, and prolongs the survival time of patients. This article reviews the advances of multi-target chimeric antigen receptor T cell, and discusses the prospect of its development.

Objective:To analyze the epidemiological, clinical characteristics and treatment of coronavirus disease 2019 (COVID-19) in Northeastern Sichuan, and summarize experience in time to provide reference for clinical diagnosis and treatment.Methods:A retrospective study was conducted. Fifty-nine patients with COVID-19 admitted to Bazhong Central Hospital and Dazhou Central Hospital from January 27th to February 9th, 2020 were selected as the subjects. The data of demography, epidemiology, laboratory examination, chest CT and related clinical treatment were collected. According to the severity of the disease, the patients were divided into three types: mild, general and severe types, and the differences of the above indices among different clinical types were compared.Results:① General information and epidemiology: 31 cases (52.5%) were male, 28 cases (47.5%) were female, the average age was (42.0±16.4) years old, and the patients over 40 years old accounted for the largest proportion (35 cases, 59.3%). The proportion of clinical type was 72.9% (43 cases) in general type, and 62.7% (37 cases) were imported type. With the increase of disease severity, the average age of patients also showed a significant increase trend [the age of the mild, general and severe patients were (30.9±13.6), (42.7±15.3), (55.8±18.9) years old, P < 0.01]. The proportion of patients with more than one basic disease in severe patients was significantly higher than those in mild and general patients [66.7% (4/6) vs. 20.0% (2/10), 9.3% (4/43), both P < 0.05]. In the distribution of clinical symptoms, the proportion of severe patients with chest distress/dyspnea was significantly higher than those in mild and general patients [66.7% (4/6) vs. 10.0% (1/10), 11.6% (5/43), both P < 0.05]. ② Laboratory examination index: the total number of white blood cell count (WBC), neutrophils count (NEU), C-reactive protein (CRP) in severe patients were higher than those in mild patients and general patients [WBC (×10 9/L): 7.21±4.35 vs. 5.85±1.69, 5.43±2.04; NEU (×10 9/L): 6.09±4.43 vs. 3.95±1.45, 3.54±1.83; CRP (mg/L): 16.00 (8.20, 46.43) vs. 5.00 (0.00, 16.13), 15.00 (3.13, 28.58)], the albumin (Alb) level in severe patients was lower than those in mild and general patients (g/L: 38.00±5.35 vs. 49.23±5.27, 39.81±2.15, both P < 0.05), while the hemoglobin (Hb) level in mild patients was higher than that in severe and general patients (g/L:155.2±12.1 vs. 141.3±6.8, 131.1±11.7, both P < 0.05). ③ Chest imaging: the CT manifestations of typical cases were single or multiple ground glass shadows. With the progress of the disease, the focus gradually increased, the scope gradually expanded, and multiple solid shadows of lung lobes were involved. ④ Treatment: all patients received at least 2 kinds of antiviral therapy, and the application rate of Interferon and Ribavirin in severe patients were higher than those in mild and general patients [100.0% (6/6) vs. 80.0% (8/10), 97.7% (42/43); 83.3% (5/6) vs. 0% (0/10), 20.9% (9/43); all P < 0.05]. ⑤ Prognosis: until March 6th 2020, 50 patients (84.8%) were discharged from the hospital after rehabilitation, and the remaining 9 patients were still under treatment, none deaths. Conclusions:The proportion of severe patients with chest distress/dyspnea is higher, the older the patients are and the more basic diseases are, the more likely they are to develop into severe type. High resolution chest CT could be considered for suspected cases or even fever patients, which may show the progress of the disease.