OBJECTIVE To standardize the main processes and related technical links of the clinical comprehensive evaluation of drugs， and provide guidance and reference for improving the quality of comprehensive evaluation evidence and its transformation and application value. METHODS The construction of Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs was based on the standard guideline formulation method of the World Health Organization （WHO）， strictly followed the latest definition of guidelines by the Institute of Medicine of the National Academy of Sciences of the United States， and conformed to the six major areas of the Guideline Research and Evaluation Tool Ⅱ. Delphi method was adopted to construct the research questions； research evidence was established by applying the research methods of evidence-based medicine. The evidence quality classification system of the Chinese Evidence-Based Medicine Center was adopted for evidence classification and evaluation. The recommendation strength was determined by the recommendation strength classification standard formulated by the Oxford University Evidence-Based Medicine Center， and the recommendation opinions were formed through the expert consensus method. RESULTS & CONCLUSIONS The Guideline for the Workflow of Clinical Comprehensive Evaluation of Drugs covers 4 major categories of research questions， including topic selection， evaluation implementation， evidence evaluation， and application and transformation of results. The formulation of this guideline has standardized the technical links of the entire process of clinical comprehensive evaluation of drugs， which can effectively guide the high-quality and high-efficient development of this work， enhance the standardized output and transformation application value of evaluation evidence， and provide high-quality evidence support for the scientific decision-making of health and the rationalization of clinical medication.

This study investigates the pharmacokinetics and metabolic characteristics of three marine-derived piericidins as potential drug leads for kidney disease: piericidin A (PA) and its two glycosides (GPAs), glucopiericidin A (GPA) and 13-hydroxyglucopiericidin A (13-OH-GPA). The research aims to facilitate lead selection and optimization for developing a viable preclinical candidate. Rapid absorption of PA and GPAs in mice was observed, characterized by short half-lives and low bioavailability. Glycosides and hydroxyl groups significantly enhanced the absorption rate (13-OH-GPA > GPA > PA). PA and GPAs exhibited metabolic instability in liver microsomes due to Cytochrome P450 enzymes (CYPs) and uridine diphosphoglucuronosyl transferases (UGTs). Glucuronidation emerged as the primary metabolic pathway, with UGT1A7, UGT1A8, UGT1A9, and UGT1A10 demonstrating high elimination rates (30%-70%) for PA and GPAs. This rapid glucuronidation may contribute to the low bioavailability of GPAs. Despite its low bioavailability (2.69%), 13-OH-GPA showed higher kidney distribution (19.8%) compared to PA (10.0%) and GPA (7.3%), suggesting enhanced biological efficacy in kidney diseases. Modifying the C-13 hydroxyl group appears to be a promising approach to improve bioavailability. In conclusion, this study provides valuable metabolic insights for the development and optimization of marine-derived piericidins as potential drug leads for kidney disease.
Animals ; Male ; Mice ; Aquatic Organisms/chemistry* ; Biological Availability ; Cytochrome P-450 Enzyme System/metabolism* ; Glucuronosyltransferase/metabolism* ; Microsomes, Liver/metabolism* ; Molecular Structure ; Biological Products/pharmacokinetics* ; Pyridines/pharmacokinetics*

ObjectiveTo investigate whether elemene（ELE） enhances the anti-glioma efficacy of cabazitaxel（CTX）， and prepare a double-targeted cationic liposome（LIP） co-loaded with ELE/CTX for the treatment of glioma， and to achieve the effect of increasing the efficacy and reducing the adverse reactions. Pharmacodynamic tests in vitro were performed to explore the advantages and mechanism of its preparation. MethodELE/CTX@LIP was prepared by high speed shear combined with probe ultrasound， the particle size and potential were characterized by nano-particle size potentiometer， and high performance liquid chromatography（HPLC） was used to determine the encapsulation efficiency and drug loading capacity of CTX/ELE. The cytotoxicity of ELE/CTX in vitro was detected by cell proliferation and activity assay（CCK8）. JMP Pro 16 software was used to optimize the process parameters of ELE/CTX@LIP based on encapsulation efficiency. The optimal cationic material type， content and ratio were screened by in vitro cytotoxicity and in vitro cell uptake， on this basis， the dual-targeted cationic liposome T7/arginine glycine aspartate tripeptide sequence（T7/cRGD）-ELE/CTX@CLIP was prepared， the stability of morphology and particle size were characterized， and the effect of T7/cRGD-ELE/CTX@CLIP on the apoptosis inducing ability and cell cycle regulation ability of glioma cells was analyzed by cell cycle and apoptosis. ResultELE/CTX showed stronger anti-glioma activity on C6 and RG2 cells. The results of in vitro cytotoxicity and in vitro cell uptake showed that the amount of cationic material was 0.10% of the total content. The optimum ratio of T7， cRGD and phospholipids was 1∶1∶50. T7/cRGD-ELE/CTX@CLIP［1，2-dilinoleyloxy-3-dimethylaminopropane（Dlin-MC3-DMA）］ and T7/cRGD-ELE/CTX@CLIP［1，2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol 2000（DMG-PEG2000）］ showed multi-level spherical nanostructures with particle sizes of 146.0， 111.3 nm， respectively， and were stable in serum. In vitro cytotoxicity results showed that T7/cRGD-ELE/CTX@CLIP had higher cytotoxicity to glioma cells than single-targeted liposomes or dual-targeted non-cationic liposomes. T7/crGD-ELE/CTX@CLIP affected the apoptosis and cycle of glioma cells， the results showed that ELE/CTX combined with liposomes could more effectively activate the apoptosis channel and inhibit the proliferation of glioma cells， and the use of T7/cRGD short peptide and cation modification enhanced the ability of apoptosis induction. ELE/CTX could effectively block glioma cell cycle at G₂/M phase， and the effect was enhanced after T7/cRGD targeted modification. ConclusionELE can enhance the anti-glioma effect of CTX. The preparation parameters of ELE/CTX@LIP are stable and feasible. Combined with the in vitro efficacy test， the anti-glioma mechanism of T7/cRGD-ELE/CTX@CLIP is preliminarily revealed.

‍ ObjectiveTo investigate the serum level of HBV RNA in untreated or treatment-experienced patients with chronic hepatitis B （CHB） and the correlation between serum HBV RNA level and the duration of antiviral therapy with nucleos（t）ide analogues （NAs）. MethodsA total of 300 patients with CHB who attended Department of Infectious Diseases in The First Affiliated Hospital of Shihezi University School of Medicine from February to July， 2022， were enrolled as subjects. Related clinical data were collected， and according to the duration of antiviral therapy， they were divided into untreated group with 73 patients， treatment duration ≤1 year group with 91 patients， and treatment duration >1 year group with 136 patients. Serum HBV RNA load， HBV DNA load， and HBsAg concentration were measured for all patients. The Mann-Whitney U test was used for comparison of continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups， further pairwise comparison using Bonferroni method； the chi-square test was used for comparison of categorical data； a Spearman correlation analysis was used to investigate the degree of correlation between various indicators. ResultsThe positive rate of HBeAg was 18.3%， and among the patients with negative HBV DNA， the patients with positive HBV RNA accounted for 44.1% （86/195）. There was a significant difference in the distribution of the serum levels of HBV RNA， HBV DNA， and HBsAg between the positive HBeAg group and the negative HBeAg group （Z=10.740， 6.300， and 7.280， all P<0.05）. There was a significant difference in the distribution of DNA level between the untreated group and the treatment duration ≤1 year group （P<0.05）； there was a significant difference in the distribution of HBV RNA and HBV DNA levels between the untreated group and the treatment duration >1 year group （P<0.05）； there was a significant difference in the distribution of HBV RNA， HBV DNA， and HBsAg levels between the treatment duration ≤1 year group and the treatment duration >1 year group （P<0.05）. The correlation analysis between the duration of antiviral therapy and the levels of HBV RNA， HBV DNA， and HBsAg showed that the duration of antiviral therapy had an extremely weak negative correlation with the levels of HBV RNA and HBsAg （r=-0.247 and -0.138， both P<0.05） and a strong negative correlation with the level of HBV DNA （r=-0.771， P<0.001）. There was a low degree of correlation between the serum level of HBV RNA and the serum levels of HBV DNA and HBsAg （r=0.360 and 0.442， both P<0.001）. Further stratified analysis showed that in the untreated group， there was a strong positive correlation between HBV RNA and HBV DNA （r=0.752， P<0.001） and a moderate positive correlation between HBV RNA and HBsAg （r=0.559， P<0.001）； in the treatment duration ≤1 year group， there was a low degree of positive correlation between HBV RNA and HBV DNA/HBsAg （r=0.396 and r=0.388， both P<0.001）； in the treatment duration >1 year group， there was a low degree of positive correlation between HBV RNA and HBsAg （r=0.352， P<0.001）. ConclusionSerum HBV RNA is negatively correlated with the duration of treatment with NAs， and the correlation of HBV RNA with HBV DNA and HBsAg gradually decreases with the increase in the duration of treatment. Therefore， it can be used as a supplementary indicator for monitoring the level of virologic response in CHB patients to a certain extent， with a relatively high accuracy in reflecting the level of viral replication in untreated patients.

OBJECTIVE To study the pharmacokinetics and tissue distribution of cannabidiol（CBD）-cholesterol succinate monoester-g-carboxymethyl chitosan （CCMC） nano-micelles in rats， and to evaluate its anti-inflammatory effect. METHODS CBD- CCMC nano-micelles were prepared by dialysis method and the properties were characterized. SD rats were divided into CBD group and CBD-CCMC nano-micelles group with 6 rats in each group. The rats were given 100 mg/kg CBD and CBD-CCMC nano- micelle by intragastric administration， respectively （based on the CBD load）. Blood was collected from the posterior ophthalmic venous plexus at 0.5， 1， 1.33， 1.5， 1.75， 2， 4， 8， 24， 48 h after administration. The heart， liver， spleen， lung， kidney and muscle tissues of rats were separated at 0.25， 1.5， 10 and 24 h after administration of CBD and CBD-CCMC nano-micelle with the same dose. The drug content in plasma and tissues was determined， the pharmacokinetic parameters were calculated， and the tissue distribution was analyzed. The inflammatory model of Caco-2 cells was induced by lipopolysaccharide， after 24 h of treatment with 5， 10， and 15 µg/mL CBD and CBD-CCMC nanomicelles （based on loaded CBD）， its anti-inflammatory activity was investigated by measuring cell viability， transepithelial electrical resistance （TEER） and inflammatory cytokines IL-1β， IL-8 and TNF-α. RESULTS The prepared CBD- CCMC nano-micelles had a particle size of （230.6±1.8） nm， a polydispersity index of 0.170±0.053， a Zeta potential of （－13.5± 1.2） mV， an encapsulation rate of （86.35±0.56）% and a drug loading of （9.18±0.32）%， respectively； the solubility was 68.240 μg/mL. The pharmacokinetic results showed that the AUC0-48 h， AUC0-∞， half-life time and peak concentration of CBD-CCMC nano- micelle group were significantly increased/extended compared with CBD group （P＜0.05 or P＜0.01）. The results of the tissue distribution study showed that at the same time point， the drug distribution concentration of CBD-CCMC nanomicelles in the rat tissue was higher than that in the CBD group. Research on anti-inflammatory effects shows that compared with CBD of the same mass concentration, CBD-CCMC nano-micelles can significantly increase cell viability （P＜0.05 or P＜0.01）， enhance TEER， and reduce the levels of IL-8， IL-1β and TNF-α in cells （P＜0.01）， and the secretion levels of inflammatory cytokines IL-8， IL-1β and TNF- α were significantly decreased （P＜0.01）. CONCLUSIONS CBD-CCMC nano-micelles can increase the plasma concentration and tissue distribution concentration of CBD， and improve anti-inflammatory activity of CBD.

Objective:To analyze the use of policy tools for rural-oriented tuition-waived medical student training policy and to provide relevant suggestions for the continuous promotion of the policy.Methods:With "rural-oriented tuition-waived medical students" as the key word, the policy texts were collected and screened from government portals. Using ROTHWELL disaggregated method to build the rural order directional medical students training policy analysis framework, applying Excel 2019 software for classification and coding of policy texts.Results:A total of 13 rural-oriented medical student training policy texts were screened and obtained. The X dimension of the policy analysis framework for rural order-oriented medical student training included three policy tools, namely, supply, environment and demand, and the Y dimension included three policy objectives, namely, available, usable, and retained. In X dimension, environmental policy tools were most frequently used. In Y dimension, the "retained" target had the highest frequency of use.Conclusion:There were differences in the frequency of using policy tools for targeted medical student cultivation in different policies. The frequency of using environmental tools is higher, which highlighted the attention of the state to medical and health services. The internal structure of policy tools is unbalanced, so the configuration of supply-oriented policy tools should be optimized, and the construction of demand-oriented policy tools should be emphasized. It is suggested to continuously optimize the combination of policy tools, improve the compatibility between policy tools and rural order-oriented medical student training, and pay attention to the sustainability of policy tools.

Objective:To establish a performance validation method for mNGS applied in BALF samples.Method:Hela cells were used as a representative of host cells, and simulated BALF samples were prepared by adding different concentrations of Hela cells, seven species of isolated pathogens (including Streptococcus pneumonia, Hemophilus influenza, Klebsiella pneumonia, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and Adenovirus), and interfering substances to sterile normal saline. Clinical BALF samples were collected simultaneously, and the results of mNGS were evaluated using traditional detection methods as a reference. The limit of detection (LOD), precision, anti-interference ability, stability, and accuracy of mNGS were determined. Results:In the simulated samples, the LOD of Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae, Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus, and Adenovirus were 150, 262, 102, 67, 96, 83 CFU/ml, and 439 copies/ml, respectively. The repeatability of the detection results for all pathogens of simulated positive BALF samples was 100%. The anti-interference test showed that the higher the concentration of human DNA, the fewer pathogen sequences detected by mNGS. Escherichia coli and Shigella sonnei were used to evaluate the ability of mNGS to distinguish closely related species. The results showed that the system could stably distinguish Escherichia coli and Shigella sonnei when the concentration of Shigella sonnei was 4, 000 CFU/ml. The stability test results showed that there was no significant change in the number of pathogen sequences detected whether after 1 to 3 freeze-thaw cycles or storage at 4 ℃, -20 ℃, or -80 ℃ for 36 h. Compared with traditional detection methods, the accuracy of 17 clinical samples was 82.4%(14/17). Continuous evaluation of clinical BALF samples simultaneously tested by mNGS and traditional methods at Tongji Hospital from October 25, 2021, to September 14, 2022, showed that the accuracy of mNGS compared to bacterial culture, fungal culture, mycobacterial culture, Mycobacterium tuberculosis culture, and conventional PCR techniques was 67.5%(472/699), 81.5%(570/699), 92.3%(335/363), 96.4%(350/363), and 86.8%(132/152), respectively. Compared with conventional PCR techniques, the accuracy of mNGS for detecting Pneumocystis jirovecii, Adenovirus, and Mycoplasma pneumoniae was 89.4%(84/94), 93.3%(56/60), and 87.1%(61/70), respectively. Conclusion:By preparing simulated BALF samples and using traditional detection methods as a reference, the performance characteristics of mNGS in detecting BALF samples can be preliminarily evaluated.

Objective:To study the epidemic status about genotype of the astrovirus, and to provide the epidemic data and control epidemic of infectious disease.Methods:Screening for astrovirus positive nucleic acids was performed using an astrovirus real-time fluorescent PCR kit. RT-PCR amplification was performed with astrovirus-specific primers Mn289/270. The positive products were recovered and purified and directly sequenced. Sequence analysis and phylogenetic analysis of astrovirus sequences were performed using Clustal and MEGA3.0 biological software.Results:From 2016 to 2018, we collected 496 fecal specimens from children with diarrhea from different regions, of them 136 cases had viral diarrhea, and in 17 of them astrovirus (3.4%) was detected, there were more males than females. All the 17 cases were under 5 years of age, and 12 cases (70.58%) were younger than 1 year old. In regional distribution, samples from Xining accounted for 64.70% (11/17), Huangzhong county for 29.41% (5/17), Huangnan Prefecture for 5.88%(1/17). The positive cases were mainly found in April to June, and there was another peak from October to December, which was consistent with the time distribution of rotavirus in past years. Seven strains were HAstV-1, one strain was HAstV-2, and the homology among four strains of HAstV-1 was 99.0%-100%. The homology with the other two strains of the same type was 88.4-95.7%.Conclusions:The dominant genotype of Astrovirus was HAstV-1 in infants with diarrhea in Qinghai, meanwhile there was also HAstV-2.

Objective:To investigate the prognosis of severe hyperbilirubinemia in full-term infants who met the exchange transfusion criteria and were treated by blood exchange transfusion and phototherapy.Methods:A total of 168 full-term infants with severe hyperbilirubinemia who met the criteria for exchange transfusion and were hospitalized in the Neonatology Department of seven tertiary hospitals in Hebei Province from June 2017 to December 2018 were retrospectively included. According to the treatment protocol, they were divided into two groups: exchange transfusion group (38 cases) and phototherapy group (130 cases). Two independent sample t-test and Chi-square test were used to compare the clinical manifestations and follow-up results between the two groups. Multivariate logistic regression was used to analyze the risk factors for poor prognosis. Results:Neonatal severe hyperbilirubinemia in the exchange transfusion and phototherapy group were both mainly caused by hemolytic disease [42.1%(16/38) and 29.2%(38/130)], sepsis [28.9%(11/38) and 11.5%(15/130)] and early-onset breastfeeding jaundice [15.8%(6/38) and 11.5%(15/130)]. Total serum bilirubin level on admission in the exchange transfusion group was significantly higher than that in the phototherapy group [(531.7±141.3) vs (440.0±67.4) μmol/L, t=3.870, P<0.001]. Moreover, the percentage of patients with mild, moderate and severe acute bilirubin encephalopathy in the exchange transfusion group were higher than those in the phototherapy group [15.8%(6/38) vs 3.8%(5/130), 7.9%(3/38) vs 0.8%(1/130), 13.2%(5/38) vs 0.0%(0/130); χ2=29.119, P<0.001]. Among the 168 patients, 135 were followed up to 18-36 months of age and 12 showed poor prognosis (developmental retardation or hearing impairment) with four in the exchange transfusion group (12.9%, 4/31) and eight in the phototherapy group (7.7%, 8/104). Multivariate logistic regression analysis showed that for full-term infants with severe hyperbilirubinemia who met the exchange transfusion criteria, phototherapy alone without blood exchange transfusion as well as severe ABE were risk factors for poor prognosis ( OR=14.407, 95% CI: 1.101-88.528, P=0.042; OR=16.561, 95% CI: 4.042-67.850, P<0.001). Conclusions:Full-term infants who have severe hyperbilirubinemia and meet the exchange transfusion criteria should be actively treated with blood exchange transfusion, especially for those with severe ABE, so as to improve the prognosis.

Objective To investigate the epidemiological characteristics and risk factors of tuberculosis in Cangzhou from 2010 to 2020. Methods A total of 325 patients diagnosed with pulmonary tuberculosis in Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2010 to December 2020 were selected as the study group. At the same time, 348 healthy adults who had a physical examination in the same hospital were selected as the control group. The epidemiological characteristics of 325 patients were analyzed. The age, gender, occupation, annual family income, marital status, drinking history, smoking history, education level and tuberculosis contact history of the two groups were compared to analyze the risk factors of tuberculosis. Results The epidemiological and statistical analyses found that among 325 cases of tuberculosis patients, patients ≥60 years old accounted for a relatively high proportion (71.08%). The majority of patients were male by gender (58.15%), and the majority of patients were farmers by occupation (47.69%). The incidence seasons were mainly in winter (40.62%) and spring (29.85%). The results of univariate analysis showed that there were statistically significant differences in gender, occupation, age, history of exposure to tuberculosis, and history of smoking between the two groups (P<0.05). Multivariate analysis found that ages ≥60 years old, male, history of smoking, farmers, and history of exposure to tuberculosis were risk factors for the onset of tuberculosis (P<0.05). Conclusion In Cangzhou, people aged over 60 years, male and farmers are prone to contract tuberculosis, and the peak time of the disease is in spring and winter. The development of corresponding prevention and control measures based on these characteristics is of great significance for the prevention and control of tuberculosis.