Detection of 14 sulfonate esters impurities of active pharmaceutical ingredients based on GC-MS/MS and LC-MS/MS / 药学学报

Two methods including gas chromatography tandem mass spectrometry (GC-MS/MS) and high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) were established to detect common alkyl sulfonates and aryl sulfonates genotoxic impurities. Four alkyl sulfonates and methyl benzenesulfonate were determined by GC-MS/MS using butyl methanesulfonate as the internal standard, the chromatographic column was HP-5MS UI (30 mm × 0.25 mm, 0.25 µm), the carrier gas was helium, the flow rate was 1.0 mL·min-1 in a constant flow mode, the sample inlet temperature was set to 250 ℃, the split ratio was 10∶1, and the initial temperature of the heating program was 80 ℃, maintained for 1 minute, and then increased to 240 ℃ at a heating rate of 30 ℃·min-1 for 2 minutes. The mass spectrometry detector was an electron bombardment ion source (EI source), the data collection condition was multi reaction monitoring mode (MRM), and method validation using the raw material of clinical drug citalopram hydrobromide as a sample. The results showed that the linear range of four alkyl sulfonates and methyl benzenesulfonate were good at 3-50 ng·mL-1 and 9-150 ng·mL-1, with a correlation coefficient of r > 0.999, The spiked recovery was 80%-120%. The detection limits were 1 and 3 ng·mL-1; Ten aryl sulfonates determined by LC-MS/MS, the chromatographic column was CSH Fluoro phenyl (100 mm × 2.1 mm, 1.7 µm), the mobile phase was methanol (B)-5 mmol·L-1 ammonium formate (D), with a flow rate of 0.2 mL·min-1, and gradient elution was performed. The gradient program (T/% B) was set as 0/20, 25/90, 35/90, 42/20. The mass spectrometer detector was electro spray ionization with positive ionization mode (ESI+), the data collection was in dynamic multi reaction monitoring mode (dMRM), and the method was validated using the raw material of the clinical drug citalopram hydrobromide as a sample. The results showed that the linear range of aryl sulfonates were good at 9-2 000 ng·mL-1, 3-100 ng·mL-1 and 0.9-30 ng·mL-1, respectively. The correlation coefficient r > 0.999, the spiked recovery was 80%-120%. The detection limits were 30, 1 and 0.3 ng·mL-1. Two detection methods did not detect potential sulfonate genotoxicity impurities in the above APIs. The established analytical methods are reliable and effective, which can provide reference for drug quality control and detection.

Growth retardation and its influencing factors in infants aged 0‒3 in Minhang District， Shanghai / 上海预防医学

ObjectiveTo investigate the detection rate and main influencing factors of growth retardation in infants aged 0-3 in Minhang District， and to provide relevant evidence for early intervention， nutrition promotion and health guidance in the future. MethodsFrom September 1， 2020 to August 31， 2021， the height， weight， basic information of parents， feeding methods， and lifestyle habits of infants who received systematic healthcare aged 0‒3 in community health service centers and Minhang maternal child health hospital were collected， and the current situation and influencing factors of infant growth retardation were analyzed. ResultsAmong the 68 637 infants who underwent a systematic physical examination in Minhang District， the total detection rate of growth retardation was 5.03% （3 453/68 637）. The detection rates in the 0-year-old， 1-year-old， 2-year-old， and 3-year-old groups were 6.57% （1 636/24 885）， 3.90% （664/17 031）， 4.62% （827/17 905）， and 3.72% （326/8 773）， respectively. There was no difference in the detection rate of growth retardation between boys and girls （P>0.05）， and a multinomial logistic regression analysis of 13 influencing factors （infant birth weight， birth length， parental weight， height， education level， mother’s childbearing age， delivery mode， household registration， feeding mode within 6 months， infant sleep， etc.） in univariate analysis showed that birth weight <2 500 g （OR=3.99， 95%CI： 2.809‒5.674） or ≥4 000 g （OR=12.78， 95%CI： 8.868‒18.443）， maternal height <150 cm （OR=7.10， 95%CI： 4.294‒11.753）， paternal height <160 cm （OR=5.65， 95%CI： 2.792‒11.422）， maternal education level of junior high school and below （OR=1.31， 95%CI： 1.087‒1.588）， paternal education level of junior high school and below （OR=1.02， 95%CI： 0.838‒1.236）， mixed feeding （OR=1.15， 95%CI： 1.031‒1.288）， and sleep duration exceeding the recommended time （OR=1.58， 95%CI： 1.466‒1.710） were risk factors for growth retardation in infants aged 0‒3. Infants with a birth length <50 cm or with household registration in Shanghai had a higher incidence of growth retardation. ConclusionGrowth retardation in infants aged 0‒3 is influenced by a combination of genetic， environmental， and sleep factors. It is essential for parents to realize the impact of growth retardation on the future of their children early on and actively participate in the early detection， screening， and intervention of growth retardation.

Etiological characteristics of viral gastroenteritis in pediatric inpatients under five years old in Shanghai， 2021‒2022 / 上海预防医学

ObjectiveTo investigate the changes in the pathogen spectrum of viral diarrhea in local pediatric inpatients as well as any variations in genotypes of major pathogens during the COVID-19 control period. MethodsFecal samples were collected from the children <5 years who were hospitalized due to acute gastroenteritis in a pediatric hospital in Shanghai. PCR test was carried out to detect rotavirus， norovirus， sapovirus， astrovirus and enteric adenovirus， and then genotyping was performed for major pathogens. ResultsOut of 546 samples， 37.55% tested positive for virus with the following positive rate ranking： norovirus GⅡ （22.16%）， group A rotavirus （16.12%）， astrovirus （2.93%）， enteric adenovirus （2.38%）， sapovirus （0.92%） and norovirus GⅠ （0.18%）. The predominant genotype within norovirus GⅡ were GⅡ.4［P31］ and GⅡ.4［P16］ with a proportion of 24.79% and 14.05% respectively. The detection rate of GⅡ.4［P31］ dropped significantly over the 2-year period （χ2＝16.140，P<0.001）. In addition， an emerging rotavirus genotype G8P ［8］， which was rarely found nationally， was discovered for the first time locally with an increasing proportion， accounting for 7.95% of all rotavirus positive cases. Phylogenic analysis demonstrated that the representative strains of this genotype were genetically closer to the DS-1-like G8P ［8］ strain found in Southeast Asia. ConclusionThe changes in the prevalence of various norovirus genotypes together with the emergence of rare rotavirus genotype in the local area illustrate the importance of continuous monitoring of viral diarrhea and genotyping of key pathogens. Increased local activity of the rare genotype also adds new parameters in the efficacy evaluation of marketed vaccines and development of potential new vaccines in near future.

Experience and Reflection on the Ethical Review of Organ Donation and Transplantation after Citizens’ Death in Medical Institution / 中国医学伦理学

In order to explore the ethical review experience of organ donation and transplantation after the death of citizens, and provide reference value for medical institutions to carry out corresponding ethical review. By using descriptive research, purpose sampling method and the principle of data saturation, 10 members and secretaries of ethics committee on clinical application of organ transplantation technology were finally selected as respondents for semi-structured interviews. The Colaizzi 7-step analysis method was adopted to analyze, summarize and refine the theme. The results showed that the ethical review experience of organ donation and transplantation after the death of citizens included four themes: the responsibilities of ethics committee, the key points of ethics review, the form of ethics review conference and its advantages and disadvantages, and the construction of the ethics committee of organ transplantation. Therefore, there are defects in the ethical review of organ donation and transplantation in medical institutions at present. These can be remedied by enriching elements of the ethical review following the four principles of medical ethics, refining the laws related to organ donation after citizens’ death, constructing a reasonable and efficient pattern of ethical review conference, and establishing a robust and appropriate operation mode of organ transplantation ethics committee.

Long-term auditory monitoring in children with Alport syndrome based on different degrees of renal injury / 临床耳鼻咽喉头颈外科杂志

Objective:To investigate long-term auditory changes and characteristics of Alport syndrome（AS） patients with different degrees of renal injury. Methods:Retrospectively analyzing clinical data of patients diagnosed AS from January 2007 to September 2022, including renal pathology, genetic detection and hearing examination. A long-term follow-up focusing on hearing and renal function was conducted. Results:This study included 70 AS patients, of which 33（25 males, 8 females, aged 3.4-27.8 years） were followed up, resulting in a loss rate of 52.9%.The follow-up period ranged from 1.1to 15.8 years, with 16 patients followed-up for over 10 years. During the follow-up, 10 patients presenting with hearing abnormalities at the time of diagnosis of AS had progressive hearing loss, and 3 patients with new hearing abnormalities were followed up, which appeared at 5-6 years of disease course. All of which were sensorineural deafness. While only 3 patients with hearing abnormalities among 13 patients received hearing aid intervention. Of these patients,7 developed end-stage renal disease（ESRD）, predominantly males （6/7）. The rate of long-term hearing loss was significantly different between ESRD group and non-ESRD group（P=0.013）. There was no correlation between the progression of renal disease and long-term hearing level（P>0.05）. kidney biopsies from 28 patients revealed varying degrees of podocyte lesion and uneven thickness of basement membrane. The severity of podocyte lesion was correlated with the rate of long-term hearing loss（P=0.048）, and there was no correlation with the severity of hearing loss（P>0.05）. Among 11 cases, theCOL4A5mutationwas most common （8 out of 11）, but there was no significant correlation between the mutation type and hearing phenotype（P>0.05）. Conclusion:AS patients exhibit progressive hearing loss with significant heterogeneity over the long-term.. THearing loss is more likely to occur 5-6 years into the disease course. Hearing abnormalities are closely related to renal disease status, kidney tissue pathology, and gene mutations, emphasizing the need for vigilant long-term hearing follow-up and early intervention.

The Effect and Mechanism of Fructus lycii on Improvement of Exercise Fatigue Using a Network Pharmacological Approach with in vitro Experimental Verification / 生物医学与环境科学（英文）

OBJECTIVE@#This study aimed to investigate the effect and underlying mechanism of Fructus lycii in improving exercise fatigue.@*METHODS@#A network pharmacological approach was used to explore potential mechanisms of action of Fructus lycii. Skeletal muscle C2C12 cells and immunofluorescence were employed to verify the effect and mechanism of the representative components in Fructus lycii predicted by network pharmacological analysis.@*RESULTS@#Six potential active components, namely quercetin, β-sitosterol, stigmasterol, 7-O-methylluteolin-6-C-beta-glucoside_qt, atropine, and glycitein, were identified to have potency in improving exercise fatigue via multiple pathways, such as the PI3K-Akt, neuroactive ligand-receptor interaction, IL-17, TNF, and MAPK signaling pathways. The immunofluorescence results indicated that quercetin, a significant active component in Fructus lycii, increased the mean staining area of 2-NBDG, TMRM, and MitoTracker, and decreased the area of CellRox compared to the control. Furthermore, the protein expression levels of p-38 MAPK, p-MAPK, p-JNK, p-PI3K, and p-AKT markedly increased after quercetin treatment.@*CONCLUSION@#Fructus lycii might alleviate exercise fatigue through multiple components and pathways. Among these, quercetin appears to improve exercise fatigue by enhancing energy metabolism and reducing oxidative stress. The PI3K-AKT and MAPK signaling pathways also appear to play a role in this process.

Proteomic analysis and validation of DNA repair regulation in the process of hepatocellular carcinoma recurrence / 临床肝胆病杂志

ObjectiveTo investigate the role and mechanism of DNA repair regulation in the process of hepatocellular carcinoma （HCC） recurrence. MethodsHCC tissue samples were collected from the patients with recurrence within two years or the patients with a good prognosis after 5 years， and the Tandem Mass Tag-labeled quantification proteomic study was used to analyze the differentially expressed proteins enriched in the four pathways of DNA replication， mismatch repair， base excision repair， and nucleotide excision repair， and the regulatory pathways and targets that play a key role in the process of HCC recurrence were analyzed to predict the possible regulatory mechanisms. The independent samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsFor the eukaryotic replication complex pathway， there were significant reductions in the protein expression levels of MCM2 （P=0.018）， MCM3 （P=0.047）， MCM4 （P=0.014）， MCM5 （P=0.008）， MCM6 （P=0.006）， MCM7 （P=0.007）， PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the nucleotide excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the base excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019） and LIG1 （P=0.042） in the HCC recurrence group； for the mismatch repair pathway， there were significant reductions in the protein expression levels of MSH2 （P=0.026）， MSH6 （P=0.006）， RFC4 （P=0.002）， RFC5 （P<0.001）， PCNA （P=0.019）， and LIG1 （P=0.042） in recurrent HCC tissue. The differentially expressed proteins were involved in the important components of MCM complex， DNA polymerase complex， ligase LIG1， long patch base shear repair complex （long patch BER）， and DNA mismatch repair protein complex. The clinical sample validation analysis of important differentially expressed proteins regulated by DNA repair showed that except for MCM6 with a trend of reduction， the recurrence group also had significant reductions in the relative protein expression levels of MCM5 （P=0.008）， MCM7 （P=0.007）， RCF4 （P=0.002）， RCF5 （P<0.001）， and MSH6 （P=0.006）. ConclusionThere are significant reductions or deletions of multiple complex protein components in the process of DNA repair during HCC recurrence.

MR-guided focused ultrasound surgery for localized prostate cancer: a preliminary investigation / 中华放射学杂志

Objective:To investigate the safety and efficacy of MR-guided focused ultrasound surgery (MRgFUS) in treating localized prostate cancer.Methods:Six patients with localized prostate cancer who underwent MRgFUS treatment from August 2020 to September 2021 in Beijing Hospital were prospectively enrolled in this study. The patients were all over 18 years old, with an average age of (68±10) years, and had not received any prior treatment for prostate cancer. Pretreatment pelvic MR and CT scans were performed to determine the region of treatment (ROT). Different urinary catheterization measures were given based on the location of the lesion. Under general anesthesia, the lesions were treated by MRgFUS using a specialized rectal ultrasound transducer on the treatment bed. The patients were followed up at 1, 3, and 6 months after treatment and annually thereafter. During follow-up, prostate-specific antigen (PSA) levels, pelvic MR scans, International Prostate Symptom Score (IPSS), International Index of Erectile Function-15 (IIEF-15) scores, and adverse events were assessed.Results:(1) All six patients underwent MRgFUS treatment for six lesions, with an average duration of (126±56) minutes, an average number of (7.3±3.2) focal ultrasound pulses per lesion, and an average non-perfusion volume of (3.8±1.1) cm 3, which covered the entire treatment target area. No treatment-related adverse events were reported. (2) The PSA levels at baseline, 1, 3, 6, and 12 months after treatment were (6.6±0.8), (3.6±1.3), (3.4±3.0), (2.5±1.7), and (2.3±1.8) ng/ml, respectively. PSA levels increased in 2 out of 6 patients during follow-up, and pelvic MR scan revealed recurrent lesions, while PSA levels continued to decrease in the remaining 4 patients, and pelvic MR scan were normal. (3) The IPSS scores at baseline, 1, and 3 months after treatment were 13.0 (4.0, 16.0), 10.0 (4.0, 12.0), and 5.0 (3.0, 6.0) points, respectively. For the three sexually active patients, the IIEF-15 scores at baseline were 40, 51, and 14 points, respectively, and IIEF-15 at 1 month after treatment were 9, 8, and 14 points, respectively, and at 3 months after treatment were 9, 66 and 26 points, respectively. (4) One patient was diagnosed with recurrence 10 months after treatment, and another patient was found to have a new lesion 6 months after treatment. Conclusions:MRgFUS might be a safe, non-invasive, and effective treatment for localized prostate cancer, but regular follow-up is vital for detecting tumor recurrence.

Research progress on cognitive impairment and intervention in patients with depression / 中华行为医学与脑科学杂志

Depression is one of the most common mental disorders, which can be accompanied by cognitive symptoms in addition to core symptoms.With the improvement of the awareness of the disease, people pay more attention to the cognitive symptoms of depression. The cognitive impairment of patients not only appears in the onset of the disease, but also persists during the remission of the disease, which has a negative impact on the patients’ social function. Some cognitive assessment tools such as cognitive tests, event-related potential(ERP) and eye movement tracking technology have been used to assess cognitive impairment in patients with depression, which show that there are multidimensional cognitive impairments. Some therapeutic methods such as exercise therapy, repetitive transcranial magnetic stimulation(rTMS) and drug therapy have shown the potential to improve the cognitive function of patients with depression. This article reviews related researches in recent years to explore the cognitive impairment and therapeutic methods of patients with depression.

Research progress on sensitivity, resistance and mechanism of plague infected animals to Yersinia pestis / 中华地方病学杂志

The sensitivity and resistance of host animals to Yersinia pestis are different among species and regions, and the indicator animals have natural resistance to Yersinia pestis. This article reviews the research progress on sensitivity, resistance and mechanism of plague infected animals to Yersinia pestis, in order to help improve the ecological research of plague and the prevention and control of plague.

Correlation of serum immunoglobulins levels with disease severity and nutritional indexes in elderly patients with heart failure / 中国医师进修杂志

Objective:To observe the correlation between serum immunoglobulins levels with the disease severity and nutritional indexes in elderly patients with heart failure.Methods:The clinical data of 53 elderly patients with heart failure (observation group) from October 2020 to January 2022 in the Affiliated Hospital of Ningbo University Medical College were retrospectively analyzed, and 53 cases of health examination at the same time were selected as the control group. The disease severity was assessed by a 6-minute walk test (6MWT), and nutritional status was assessed by nutritional index. The serum levels of IgA, IgM and IgG were measured by PUZS-300 fully automated biochemical analyzer; the serum nutrient indexes were measured by enzyme linked immunosorbent assay, including total protein, albumin and proalbumin. Pearson method was used for correlation test; the value of IgA, IgM and IgG in predicting disease severity in elderly patients with heart failure was analyzed by the receiver operating characteristic (ROC) curve.Results:The IgA, IgM and IgG in observed group were significantly higher than those in control group: (4.31 ± 1.50) g/L vs. (1.63 ± 0.71) g/L, (18.50 ± 3.93) g/L vs. (11.33 ± 1.12) g/L and (4.58 ± 1.26) g/L vs. (1.96 ± 0.36) g/L, and there were statistical differences ( P<0.01). The 6MWT result showed that 23 cases were with mild status, 18 cases moderate, and 12 cases severe. The IgA, IgM and IgG in moderate and severe patients were significantly higher than those in mild patients: (4.34 ± 0.78) and (6.38 ± 0.91) g/L vs. (3.23 ± 0.94) g/L, (18.90 ± 2.09) and (23.81 ± 2.78) g/L vs. (15.44 ± 2.03) g/L, (4.77 ± 0.56) and (6.47 ± 0.34) g/L vs. (3.45 ± 0.44) g/L, the indexes in severe patients were significantly higher than those in moderate patients, and there were statistical differences ( P<0.05). Pearson correlation analysis result showed that IgA, IgM and IgG were positively correlated with disease severity in elderly patients with heart failure ( r = 0.801, 0.830 and 0.925; P<0.01). The nutritional index result showed that malnutrition was in 25 cases, and normal nutrition in 28 cases. The IgA, IgM and IgG in patients with malnutrition were significantly higher than those in patients with normal nutrition: (4.89 ± 0.80) g/L vs. (1.98 ± 0.69) g/L, (19.45 ± 2.01) g/L vs. (12.01 ± 1.28) g/L and (4.88 ± 0.59) g/L vs. (2.01 ± 0.47) g/L, the nutritional index, total protein, albumin and proalbumin were significantly lower than those in patients with normal nutrition: (49.44 ± 6.98) scores vs. (58.34 ± 8.12) scores, (62.87 ± 5.20) g/L vs. (66.74 ± 4.54) g/L, (33.21 ± 2.64) g/L vs. (39.12 ± 3.11) g/L and (185.55 ± 16.72) mg/L vs. (196.33 ± 18.90) mg/L, and there were statistical differences ( P<0.01 or <0.05). Pearson correlation analysis result showed that the IgA and IgM were negatively correlated with nutritional index, total protein, albumin and proalbumin in elderly patients with heart failure (IgA: r = - 0.456, - 0.353, -0.620 and -0.281; IgM: r = -0.445, -0.327, -0.611 and -0.321; P<0.01 or <0.05); IgG was negatively correlated with nutritional index, total protein and albumin ( r = - 0.425, - 0.359 and - 0.646; P<0.01), and it had no correlation with proalbumin ( P>0.05). ROC curve analysis result showed that the areas under the curve of IgA, IgM and IgG in predicting disease severity in elderly patients with heart failure were 0.891, 0.943 and 0.838, sensitivity of 76.70%, 90.00% and 73.33%, and specificity of 87.00%, 87.00% and 100.00%. Conclusions:The serum levels of IgA, IgM and IgG increased significantly in elderly patients with heart failure, who were also with abnormal immune function and decreased nutritional status. The serum levels of IgA, IgM and IgG correlate with disease severity and nutritional indexes.

Observation on the effect of power PICC with two different external lengths in catheterization of patients with hematologic diseases / 中国实用护理杂志

Objective:To investigate the effect of power PICC with two different external lengths in patients with hematologic diseases.Methods:From August 2020 to August 2021, a retrospective analysis was conducted on 233 cases of hematologic diseases who received power PICC catheterization in Department of Hematology, the Nanfang Hospital of Southern Medical University, the patients were divided into 0-1 cm group (132 cases) and 2-4 cm group (101 cases) according to the external lengths after catheter insertion. The incidence of related complications and patient comfort at 24 hours, 72 hours, 7 days and 14 days after catheterization were compared between the two groups.Results:At 24 hours and 72 hours after catheterization, the incidence of bleeding at puncture point in 0-1 cm group was 14.39%(19/132) and 9.85%(13/132), in 2-4 cm group was 30.69%(31/101) and 32.67%(33/101), the difference was statistically significant ( χ2=9.02, 14.16, both P<0.05). The rate of redness and swelling of punture point at 7 days after catheterization was 9.09%(12/132) and 40.59%(41/101) in the two groups, with statistical significance ( χ2=32.32, P<0.05). The rates of slightly prolapse at 7 and 14 days after catheterization were 18.18% (24/132) and 18.94%(25/132) in 0-1 cm group, 59.41% (60/101) and 67.33%(68/101) in 2-4 cm group, the difference between the two groups was statistically significant ( χ2=42.18, 55.86, both P<0.05). In the observation of comfort at 14 days after catheterization, the comfort score of patients in the 0-1 cm group was significantly higher than that in the 2-4 cm group, and the difference between the two groups was statistically significant ( χ2=7.34, P<0.05). Conclusions:Patients with hematologic diseases can reduce the risk of bleeding and slightly prolapse, and improve the comfort of patients when the external lengths is 0-1 cm of power PICC, which is worthy of clinical application and promotion. The catheterizer should adopt accurate in appropriate measurement method to determine the best lengths of the catheters.

Correlation between white matter hyperintensities and outcome after intravenous thrombolysis in patients with acute ischemic stroke / 国际脑血管病杂志

Objective:To investigate the correlation between different degrees of white matter hyperintensities (WMHs) and outcome after intravenous thrombolysis (IVT) in patients with acute ischemic stroke.Methods:Patients with AIS received intravenous thrombolytic therapy with standard dose of alteplase in the First Hospital of Putian City from January 2019 to June 2022 were retrospectively included. The Fazekas scale was used to score the WMHs shown on MRI, and the patients were divided into without or mild WMH group and moderate-to-severe WMH group. The clinical baseline data and the clinical outcome after 3 months of the two groups were compared. The poor outcome was defined as the modified Rankin scale score >2. Multivariate logistic regression analysis was used to determine the influence of the severity of WMHs on the outcome after intravenous thrombolysis. Results:A total of 103 patients with AIS were included. Their age was 64.85±10.89 years old, and 66 (64.1%) were men. There were 60 patients (58.3%) in the without or mild WMH group, and 43 (41.7%) in the moderate-to-severe WMH group. There were significant differences in age, body mass index, systolic blood pressure, baseline National Institutes of Health Stroke Scale (NIHSS) scores, as well as the proportion of patients with hypertension, smoking, hemorrhagic transformation and poor functional outcome at 3 months after onset in different degrees of WMH groups (all P<0.05). There were 73 patients (70.9%) in the good outcome group and 30 (29.1%) in the poor outcome group. There were significant differences in age, body mass index, baseline NIHSS score, WMH score, as well as the proportion of patients with hypertension, large atherosclerotic stroke and symptomatic intracranial hemorrhage between the two groups ( P<0.05). Multivariate logistic regression analysis showed that after adjusting for confounding factors, moderate-to-severe WMHs were the independent risk factors for the poor outcome at 3 months after intravenous thrombolysis (odds ratio 3.810, 95% confidence interval 1.298-1.124; P=0.015). Conclusion:Moderate-to-severe WMHs are associated with the poor outcome in patients with AIS at 3 months after intravenous thrombolysis.

Historical Tracing and Textual Research on Ancient Classic Formula Zhenwutang / 中国实验方剂学杂志

ZHANG Zhongjing's Zhenwutang is a classic formula for warming Yang and excreting water. It is composed of Aconiti Lateralis Radix Praeparata， Poria， Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens. Physicians of later generations have inherited and developed this formula by detailed recording and application. This paper adopted bibliometrics method to analyze Zhenwutang in terms of history， indications， dosage， drug processing， usage， and modification. The results showed that Zhenwutang was most widely used in Ming and Qing dynasties. Many physicians have inherited ZHANG Zhongjing's theory regarding the application of Zhenwutang in disease treatment， and a few physicians have used it to treat diphtheria and water-related diseases. Some physicians modified this formula to treat maculae， intermittent dysentery， jaundice and so on. Zhenwutang was mainly used to treat diseases of the circulatory system， respiratory system and urinary system in modern clinical practice. The processing of herbal medicines in this formula was clear. Specifically， the raw material of Aconiti Lateralis Radix Praeparata needed to be processed and peeled， while those of Poria， Atractylodis Macrocephalae Rhizoma， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens can be used directly. Although being different， most of the dosages were consistent with those in Treatise on Febrile Diseases. According to the textual research， it is suggested that the reference dosage of this prescription in clinical practice is 41.25 g for Poria， Paeoniae Radix Alba， and Zingiberis Rhizoma Recens， respectively， 27.5 g for Atractylodes macrocephala， and 15 g for Aconiti Lateralis Radix Praeparata. The medicinal materials should be decocted in 1 600 mL water to reach a volume of 600 mL. After removal of the residues， the decoction should be taken warm with 140 mL each time， three times a day. The textual research of Zhenwutang is expected to provide a theoretical reference for the clinical application and formulation of Zhenwutang.

Role and possible mechanism of estrogen receptor α down-regulation leading to damage of TM4 Sertoli cell connectivity in mice / 中国药理学通报

Aim To investigate the role of autophagy in the dysfunction of testicular TM4 cell junction induced by ERα down-regulation. Methods TM4 cells were treated with different concentrations of E R a inhibitor ICI182780 (ICI), and the proliferative activity of TM4 cells was detected by CCK-8 method. The number and morphological changes of TM4 cells were observed by light microscope. The levels of E R a, junction function related proteins and autophagy marker proteins were detected by Western blot. The expression and localization of Cx43 were detected by immunofluorescence staining. The cells were treated with chloroquine (CQ) and ICI for 24 h. The expression levels of autophagy and junction function related proteins were detected by Western blot. Results When ICI concentration was 50 nmol • L ~ or above, the cell viability decreased significantly. The increase of cell vacuoles in ICI group was observed by light microscope. Compared with normal control group, the protein expression levels of E R a, ZO-1, occludin, claudin-11, p-catenin and Cx43 in ICI groups significantly dropped, while the expression levels of N-cadherin and E-cadherin had no significant changes; LC3 II significantly rose, while p62 expression significantly fell. The results of immunofluorescence showed that the fluorescence expression of Cx43 in ICI group decreased significantly, but the position of CX43 did not change significantly. Compared with ICI group, the expression levels of LC3 II, p62, Cx43, ZO-1 and β-Catenin significantly increased. Conclusions The down-regulation of E R a leads to damage of TM4 cell junction function, which may be related to the activation of autophagy.

Effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress / 中国药理学通报

Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress. Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly, with six mice in each group. The mice in normal group or high-fat diet group were fed with regular or high-fat diet continuously for five months. The mice were weighed, anesthetized, and euthanized to collect testicular and epididymal tissue for analysis. The testicular tissue was weighed and their indices were calculated. Epididymal tissue was collected for semen analysis. The morphological alterations of testicular tissue were observed using hematoxylin-eosin ( HE ) staining. The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calculated. The expression levels of apoptosis and endoplasmic reticulum stress-related proteins in testicular tissue were detected by Western blot. The protein expression and localization of GRP78 in testicular tissue were further detected by immunofluorescence. Results The results showed that compared to the normal group, the high-fat diet group had a significant increase in body weight, a significant decrease in testicular index, sperm concentration, and sperm vability, loose arrangement of germ cells, significant thinning of the seminiferous epithelium, no significant change in the diameter of seminiferous tubules, a significant increase in germ cell apoptosis , with an increased apoptosis index, and significant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expression. The expression levels of GRP78 , p-IREl, XBP1, and ATF6a proteins were significantly up-regulated, while p-PERK, p-eIF2a, ATF4 protein expression showed no significant changes. Immunofluorescence results further showed a significant increase in the expression of GRP78 protein in the testicular tissue,with no significant changes in the expression location. Conclusions High-fat diet can induce the apoptosis of mouse testicular germ cells, and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.

Analysis of clinical effect and influencing factors of history-indicated cerclage or ultrasound-indicated cerclage in singleton pregnancy / 中华妇产科杂志

Objective: To investigate the clinical effect and the influencing factors of ultrasound-indicated cerclage and history-indicated cerclage in singleton gestation. Methods: The clinical data of 272 singleton pregnant women with cervical incompetence who underwent McDonald cervical cerclage due to medical history indication (history-indicated group) or ultrasound indication (ultrasound-indicated group) in Peking University First Hospital from January 2010 to February 2021 were retrospectively analyzed. The general clinical data and maternal and fetal outcomes were compared between the history-indicated group (141 cases) and ultrasound-indicated group (131 cases). According to the gestational age at delivery, 272 pregnant women who underwent cervical cerclage were further divided into ≥34 weeks group (225 cases) and <34 weeks group (47 cases), and the influencing factors of preterm birth before 34 weeks of gestation were analyzed. Results: (1) The median gestational age at cerclage was 16.6 weeks in the history-indicated group and 23.4 weeks in the ultrasound-indicated group, and the median gestational age extension at delivery was 21.4 weeks and 14.7 weeks, respectively, with statistically significant differences between the two groups (all P<0.05). (2) The full-term birth rate was 76.6% (108/141) in the history-indicated group and 71.0% (93/131) in the ultrasound-indicated group, the live birth rate was 97.2% (137/141) and 97.7% (128/131), and the median birth weight of live birth was 3 155 g and 3 055 g, respectively. The differences were not statistically significant (all P>0.05). Among 272 pregnant women with cervical cerclage, 265 neonates survived (97.4%, 265/272). The gestational age of 7 pregnant women who did not have live birth was ≤25 weeks of gestation (range: 19+1-25 weeks), and they were all clinically infected or confirmed chorioamnionitis or pathogenic microorganisms carrying during pregnancy, and their families gave up. The minimum birth weight of the surviving neonate was 850 g (gestational week of delivery was 26+6 weeks). (3) Univariate analysis showed that compared with ≥34 weeks group, the body mass index (BMI) of pregnant women in <34 weeks group was higher at 6-7 weeks of gestation (median: 24.5 vs 25.4 kg/m2), shorter cervical length (CL) at 1-2 weeks after surgery [(31.1±8.4) vs (26.1±11.0) mm], shorter CL at 26-28 weeks of gestation after surgery (median: 26.3 vs 16.0 mm), and higher incidence of elevated C-reactive protein (CRP) before and after surgery and before delivery. The differences were all statistically significant (all P<0.05). Multivariate logistic regression analysis showed that preterm birth before 34 weeks was negatively associated with CL at 26-28 weeks of gestation after cerclage (OR=0.902, 95%CI: 0.858-0.947; P<0.001), and was positively correlated with elevated CRP before delivery (OR=3.492, 95%CI: 1.652-7.381; P=0.001). There were no significant correlations between preterm birth and preoperative or postoperative CRP elevation, CL at 1-2 weeks after surgery, and BMI at 6-7 weeks of gestation (all P>0.05). Conclusions: Cervical cerclage for singleton pregnant women with cervical incompetence indicated by history or ultrasound both have good clinical efficacy, and there is no significant difference in maternal and fetal outcomes between the two groups. CL at 26-28 weeks of gestation and CRP before delivery are risk factors for preterm birth before 34 weeks of gestation after cervical cerclage.

Comparison of BCR::ABL (P210) mRNA levels detected by dPCR and qPCR methods in patients with chronic myeloid leukemia / 中华血液学杂志

Objective: To compare digital polymerase chain reaction (dPCR) and real-time quantitative PCR (qPCR) measurements of BCR::ABL (P210) mRNA expression in patients with chronic myeloid leukemia (CML) . Methods: In this non-interventional, cross-sectional study, BCR::ABL (P210) mRNA was simultaneously measured by dPCR and qPCR in peripheral blood samples collected from patients with CML who underwent tyrosine kinase inhibitor therapy and who achieved at least a complete cytogenetic response from September 2021 to February 2023 at Peking University People's Hospital. The difference, correlation, and agreement between the two methods were evaluated using the Wilcoxon signed-rank test, Spearman's correlation, and Bland-Altman analysis, respectively. Results: In total, 459 data pairs for BCR::ABL mRNA expression measured by dPCR and qPCR from 356 patients with CML were analyzed. There was a significant difference in BCR::ABL mRNA expression between the two methods (P<0.001). When analyzed by the depth of the molecular response (MR), a significant difference only existed for patients with ≥MR4.5 (P<0.001). No significant difference was observed for those who did not achieve a major MR (no MMR; P=0.922) or for those who achieved a major MR (MMR; P=0.723) or MR4 (P=0.099). There was a moderate correlation between the BCR::ABL mRNA expression between the two methods (r=0.761, P<0.001). However, the correlation gradually weakened or disappeared as the depth of the MR increased (no MMR: r=0.929, P<0.001; MMR: r=0.815, P<0.001; MR4: r=0.408, P<0.001; MR4.5: r=0.176, P=0.176). In addition, the agreement in BCR::ABL mRNA expression between the two methods in those with MR4.5 was weaker than other groups (no MMR: ▉= 0.042, P=0.846; MMR:▉=0.054, P=0.229; MR4:▉=-0.020, P=0.399; MR4.5:▉=-0.219, P<0.001) . Conclusions: dPCR is more accurate than qPCR for measuring BCR::ABL (P210) mRNA expression in patients with CML who achieve a stable deep MR.

Advanced subunit vaccine delivery technologies: From vaccine cascade obstacles to design strategies

Designing and manufacturing safe and effective vaccines is a crucial challenge for human health worldwide. Research on adjuvant-based subunit vaccines is increasingly being explored to meet clinical needs. Nevertheless, the adaptive immune responses of subunit vaccines are still unfavorable, which may partially be attributed to the immune cascade obstacles and unsatisfactory vaccine design. An extended understanding of the crosstalk between vaccine delivery strategies and immunological mechanisms could provide scientific insight to optimize antigen delivery and improve vaccination efficacy. In this review, we summarized the advanced subunit vaccine delivery technologies from the perspective of vaccine cascade obstacles after administration. The engineered subunit vaccines with lymph node and specific cell targeting ability, antigen cross-presentation, T cell activation properties, and tailorable antigen release patterns may achieve effective immune protection with high precision, efficiency, and stability. We hope this review can provide rational design principles and inspire the exploitation of future subunit vaccines.

Advances of metal nanoadjuvant for cancer immunotherapy / 药学学报

Immunotherapy has become another effective tumor treatment after surgical resection, chemotherapy, radiotherapy and targeted therapy. However, due to the low immunogenicity of tumor cells and immunosuppressive tumor microenvironment, antigen-presenting cells inefficiently process and present tumor antigens, thus leading to insufficient activation of cytotoxic T lymphocytes and tumor infiltration, which significantly affects the effectiveness of tumor immunotherapy. In recent years, it has been demonstrated that multiple metal ions exhibit distinguished modulatory effects in activating innate immune stimulation and conquering acquired immune tolerance. Based on this, scientists have designed a series of nano-adjuvant delivery systems with metal ions or metal nanoparticles to enhance the targeted accumulation of metal ions in tumor tissues or lymphoid organs for efficiently inducing immunogenic cell death or directly activating antigen-presenting cells to initiate anti-tumor specific immune response. This review briefly outlines the role of various metal ions in anti-tumor immunomodulation, summarizes the research progress in using metal nanoadjuvant delivery systems to achieve efficient anti-tumor immunotherapy, and provides foresight on the main challenges and potential directions in this field.