To establish an LC-MS/MS method for simultaneous determination of peimine and peiminine in rat plasma after oral and intravenous administration of Fritillaria thunbergii Miq. extract, the pharmacokinetic parameters were calculated as well. Peimine, peiminine and internal standard carbamazepine were extracted from plasma with liquid-liquid extraction by ethyl acetate, then separated on a Luna C18 column by using acetonitrile-water containing 10 mmol x L(-1) ammonium formate (35:65), as mobile phase. The electrospray ionization (ESI) source was applied and operated in positive ion mode. Peimine was detected at m/z 432.4 --> 414.4, peiminine at m/z 430.4 --> 412.4 and carbamazepine (IS) at 237.1 --> 194.2. The linear calibration curves were obtained at the concentration range of 0.8-800 ng x mL(-1) for peimine and peiminine. The extraction recoveries were 94.1%-105.3% and 85.8%-98.6%, respectively. The precisions, accuracy and stability of the analytes meet the requirements. The method was shown to be effective, convenient, and suitable for simultaneous pharmacokinetic study of peimine and peiminine in rat.

Objective To investigate the inhibitory effect of IN5 from chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia trachomatis (Ct).Methods PCR was used to amplify IN5 gene from Vp1 DNA of phiCPG1, then the recombinant plasmid pET28a/IN5 was constructed.After transformation, the fusion protein IN5 was induced,identified and purified.Ct was incubated with the purified IN5 protein or Vp1 protein.After 48 h of incubation, the inclusion bodies were counted with iodine staining and indirect immunofluorescence.One-way ANOVA was used to compare the difference of inclusion bodies among groups.If the difference among the groups was statistically significant, the Bonferroni method was used to compare any two mean values.Finally, the inhibitory rate of IN5 protein and Vp1 protein to Ct was calculated.Results IN5 protein from chlamydiaphage phiCPG1 capsid protein Vp1 was successfully obtained.At the same concentration of 53μg/mL,the inhibitory rates of Ct growth in IN5 and in Vp1 groups were 52.42% and 78.04%, respectively.Conclusion IN5 protein has inhibitory effect on the growth of Ct,but the inhibitory rate is lower than that of Vp1, which provides a preliminary clue for searching the dominant region of Vp1 protein inhibiting the growth of Ct.

Objective To determine the potential correlation of 5-HTT gene polymorphisms (5-HTTLPR and STin2) with clinical manifestations in depression.Methods A total of 401 depressed patients,all of Chinese Han region,were collected and genotyped by polymerase chain reactions (PCR).All patients were evaluated using a 17-item Hamilton depression rating scale (HAMD-17) and Hamilton anxiety rating scale (HAMA),and then associated analysis was applied.Results (1) The age of onset in patients with L/S genotype of 5-HTLPR polymorphism were much younger than that of patients with L/L and S/S genotype (F=3.281,P=0.039).Besides,there was also a significant difference of HAMA1 (anxious mood) scores among patients with different genotypes for 5-HTTLPR polymorphism,where the scores of those with L/S genotype were the highest (2.34±0.80,P=0.010).(2) The scores of HAMD10 (mental anxiety),HAMA1 (anxious mood),HAMA3 (fear) and mental anxiety factor were higher in patients with 12/10 genotype than patients with 12/12 and 10/10 genotype for STin2 polymorphism (2.40±0.83,2.38±0.90,1.42± 1.04,14.60±4.26 respectively;P value:0.014,0.044,0.03 and 0.006 respectively).The scores of HAMD10(mental anxiety) and mental anxiety factor were (2.11±0.77),(12.96±3.78) in the 12/12 genotype patients,and significantly lower than that in the 12/10 genotype patients (adjustedPvalue:0.018,0.006).Conclusions A positive association of the 5-HTT polymorphisms with anxious symptoms in depressed patients is revealed.These findings might provide some evidences for the clinical phenotype and optimization of depression treatment.

Abstract: Objective To investigate the clinical features and risk factors for severe tsutsugamushi disease, so as to provide reference for diagnosis and differentiation of severe tsutsugamushi disease as soon as possible. Methods The clinical data of 178 cases of inpatients with tsutsugamushi disease admitted to the Guangzhou Eighth People's Hospital, Guangzhou Medical University from January 2016 to September 2021 were collected and analyzed according to their gender, age, underlying diseases, clinical characteristics at admission, laboratory examination results within 24 hours of admission and epidemiological history. The patients were divided into the severe group and the non-severe group according to the diagnostic criteria. The data of clinical characteristics, laboratory examination and prognosis of the two groups were compared. Multivariate logistic regression analysis was performed on the variables with statistical significance and the receiver operating characteristic curve (ROC) was drawn. Results A total of 178 patients were included in this study, with 37 in the severe group and 141 in the non-severe group. Compared with the non-severe group, the age of the severe group was older, the underlying diseases were more, the incidence of dyspnea and the levels of white blood cell, total bilirubin, aspartate aminotransferase, lactate dehydrogenase, cystatin C, uric acid and serum creatinine were significantly increased, the levels of platelet and albumin were significantly decreased (all P<0.05). The dyspnea [odds ratio (OR value)=8.93, 95% confidence interval (CI): 1.200-66.424; P=0.032], total bilirubin (OR=1.091, 95%CI: 1.028-1.159; P=0.004) and serum creatinine (OR=1.052, 95%CI: 1.004-1.102; P=0.033) were independent risk factors for severe tsutsugamushi disease. The area under ROC curve of total bilirubin and serum creatinine were 0.777 and 0.764, respectively (both P<0.01), indicating high predictive value for severe tsutsugamushi disease. The optimal cut-off value for total bilirubin was 23.01 µmol/L, with a sensitivity of 54.10% and a specificity of 90.60%; the optimal cut-off value for creatinine was 126.45 µmol/L, with a sensitivity of 43.20% and a specificity of 100.00%. The case fatality rate of severe tsutsugamushi disease was 2.70%. Conclusions The patients with severe tsutsugamushi disease are older, and have more underlying diseases. Dyspnea, increased total bilirubin and elevated serum creatinine are independent risk factors for severe tsutsugamushi disease, which can help in the early identification of severe tsutsugamushi disease early.

We wished to undertake molecular genetic typing and evaluate recombinants of the hepatitis-B virus (HBV) in Tibet (China). Multistage random sampling was used to collect HBsAg-positive samples. Nested polymerase chain reactions were used to amplify the whole sequence of the HBV. DNAstar, MEGA6 and SimPlot were used to assemble sequences, create phylogenetic trees, and undertake recombination analyses. Twelve whole sequences of the HBV of a Tibetan population were collected using these methods. Results showed that all 12 strains were C/D recombinants. Nine of the recombinations were at nt750, and the other three at nt1526. Therefore, the 12 strains could be divided into two types of recombinants: C/Da and C/Db. Analyses of the sequence of the whole genome revealed that the 12 strains belonged to genotype C, and that the nucleotide distance was > 4% between the 12 strains and sub-genotypes C1 to C15 in Genbank. The most likely sub-genotype was C1. Individuals with C/Da were from central and northern Tibet (e.g., Lasa, Linzhi, Ali) and those with C/Db recombinants were from Shannan in southern Tibet. These data suggest that the two types of recombinants had a good distribution in Tibet. Also, they can provide important information for studies on HBV recombination, gene features, virus evolution, as well as the control and prevention of HBV infection in Tibet.
Adult ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Tibet ; Young Adult

Starting from February 2014, the Ebola virus outbreak had spread across West African countries within a few months and caused great concerns of the World Health Organization. Currently no effective vaccines or drugs have been available for prevention and treatment of Ebola virus infection. This paper gives a brief review of the epidemics and pandemics, the biological characteristics of Ebola virus, the potential antiviral drug targets, and research progress of vaccine and drug development against the virus.
Antiviral Agents ; therapeutic use ; Ebolavirus ; Hemorrhagic Fever, Ebola ; prevention & control ; therapy ; Humans

Objective To evaluate the effects of interleukin-22 (IL-22) on the expression of CC chemokine ligand 27 (CCL27) in human epidermal keratinocytes,and to explore its mechanism.Methods Immunohistochemical study was performed to determine the expression of CCL27 in skin lesions of 10 patients with psoriasis and skin tissues of 5 healthy controls.Cultured HaCaT cells were divided into 8 groups:control group treated with PBS,5 IL-22 groups treated with 12.5,25,50,100 and 200 μg/L IL-22 respectively,2 signaling pathway inhibition groups treated with 50 μrmol/L AG490 (JAK2/STAT3 signaling pathway inhibitor) or PD98059 (MAPK-ERK1/2 signaling pathway inhibitor) for 2 hours followed by the treatment with 50 μg/L IL-22 in the 5％ CO2 incubator at 37 ℃.After 24-hour cultivation,total proteins were extracted,and culture supernatants were collected,and both Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to determine the expression of CCL27.Results Immunohistochemical study showed that the expression of CCL27 was significantly higher in the skin lesions of the patients with psoriasis than in the skin tissues of the healthy controls.Western blot analysis revealed that the protein expression of CCL27 in the 12.5-,25-,50-,100-and 200-μg/L IL-22 groups was 0.491 ± 0.013,0.620 ± 0.019,0.623 ± 0.014,0.802 ± 0.052 and 1.138 ± 0.013 respectively,which were all higher than that in the control group (0.413 ± 0.013,all P ＜ 0.01).The expression of CCL27 was significantly lower in the IL-22 + AG490 group (0.411 ± 0.019) and IL-22 + PD98059 group (0.280 ± 0.012) than in the 50-μg/L IL-22 group (both P ＜ 0.01).ELISA also showed the same trend of changes in the level of CCL27 in the above groups as Western blot showed.Conclusion IL-22 can promote the expression of CCL27 in HaCaT cells,which may be associated with the MAPK-ERK 1/2 and JAK2/STAT3 signaling pathways.

ObjectiveTo analyze the on-site technical evaluation results of the provincial-level health enterprises in Shandong Province. Methods A total of 88 provincial-level health enterprises in Shandong Province in 2021 were selected as the research subjects using the convenient sampling method. The scores of on-site technical assessments were analyzed. Results The total score of on-site technical evaluations for the 88 provincial health enterprises was (942.1±21.8) points. The scores for the first-level indicators, including management system, health environment, health management and services, and health culture, were (193.7±4.7), (191.6±5.2), (414.4±16.4), and (142.3±6.7) points, respectively. The score for health culture in large-sized enterprises was higher than that in medium-sized and below enterprises (P<0.05). State-owned enterprises had higher total scores than private enterprises and joint-stock enterprises (both P<0.05). The score for health management and services in the mining industry was higher than that in the manufacturing industry (P<0.05). Enterprises with high total investment in health promotion had higher scores for health culture than those with low investment (P<0.05). There were no significant differences in the scores for indicators of management system and health environment among enterprises of different sizes, natures, industries, and total investments in health promotion (all P>0.05). Conclusion The overall score of these 88 enterprises was higher than the standard for provincial health enterprises. Except for management systems and general environment, there were some differences in the scores for health management and services, and health culture assessments among enterprises of different sizes, natures, industries, and total investments in health promotion.

Objective:To explore the clinical characteristics of imported malaria from Africa, and the correlation between blood lipids and disease severity.Methods:The clinical data of 172 imported malaria patients from Africa were collected from January 1, 2018 to January 31, 2021 in Guangzhou Eighth People′s Hospital, Guangzhou Medical University. The general conditions, clinical symptoms, laboratory tests, treatment and prognosis of the patients were analyzed retrospectively. The independent samples t-test, Mann-Whitney U test and binary multivariate logistic regression analysis were used for statistical analysis when appropriate. Results:A total of 172 patients were divided into 39 severe cases and 133 non-severe cases in this study. The main infection species was Plasmodium falciparum, with a total of 153 cases (89.0%), and the severe cases were all falciparum malarias. Compared with the non-severe group, the white blood cell count and triacylglycerol level were increased in the severe group.The differences were both statistically significant ( Z=2.397 and 4.368, respectively; both P<0.05). The hemoglobin, hematocrit, platelet count, high-density lipoprotein cholesterol, apolipoprotein A (ApoA)Ⅰ and ApoAⅠ/apolipoprotein B (ApoB) were significantly decreased in the severe group. The differences were all statistically significant ( Z=-4.473, -4.464, -4.750, -4.826, -5.488 and -4.419, respectively; all P<0.01). The total cholesterol and low-density lipoprotein cholesterol were also reduced in the severe group. The differences were both statistically significant ( t=3.817 and 5.285, respectively; both P<0.01). The area under receiver operator characteristic curve of high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triacylglycerol, ApoAⅠ, and ApoAⅠ/ApoB were 0.754, 0.727, 0.730, 0.789 and 0.733, respectively (all P<0.01). When the Youden index was at its maximum, the best cut-off value of ApoAⅠ was 0.535 g/L with sensitivity of the prediction for severe malaria of 79.5% and the specificity of 68.4%. ApoAⅠ had independent predictive value for severe malaria (odds ratio ( OR)=0.013, 95% confidence interval ( CI) 0.002 to 0.086, P<0.01), and stratified according to the best cut-off value, the risk of severe malaria when ApoA Ⅰ<0.535 g/L was 8.396 times of ApoA Ⅰ≥0.535 g/L ( OR=8.396, 95% CI 3.557 to 19.820, P<0.01). The case fatality rate of severe malaria was 2.6%(1/39). Conclusions:The imported malaria patients from Africa are mainly infected with Plasmodium falciparum and the case fatality rate of severe malaria is high. The high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triacylglycerol, ApoAⅠ, ApoAⅠ/ApoB of blood lipids show higher predictive value for severe malaria, among which, ApoAⅠ has independent predictive value for severe malaria. Changes in blood lipids will help to identify severe malaria as early as possible, improve the cure rate, and reduce the risk of death.

Objective:To analyze the epidemiological and clinical characteristics of patients with tsutsugamushi disease in Guangdong Province from 2016 to 2021, and to provide a reference for clinical diagnosis, treatment, scientific prevention and control of tsutsugamushi disease.Methods:A retrospective analysis was conducted to collect the information of patients with tsutsugamushi disease admitted to Guangzhou Eighth Hospital Affiliated to Guangzhou Medical University from 2016 to 2021, including the basic information, epidemiological characteristics, clinical manifestations, auxiliary examinations, complications, misdiagnosis at first diagnosis and treatment outcomes.Results:Among 155 patients with tsutsugamushi disease, there were 75 males (48.39%) and 80 females (51.61%), with an e age of (54.41 ± 13.78) years old, and 30 cases (19.35%) had other underlying diseases. The peak time of onset was from June to September. There were 97 local patients (62.58%) in Guangzhou, and 58 cases (37.42%) in other prefecture-level cities; 76.77% (119/155) had a history of field activities before the onset of the disease. 36.13% (56/155) were farmers. The most common clinical manifestations were fever (100.00%, 155/155), chills and/or shivering (77.42%, 120/155), headache and/or dizziness (74.19%, 115/155), fatigue (65.81%, 102/155), eschar or ulcer (92.90%, 144/155), and lymphadenopathy (49.68%, 77/155). The laboratory test results mainly showed a decrease in eosinophils (81.94%, 127/155), a decrease in hematocrit (78.71%, 122/155), a decrease in hemoglobin (52.26%, 81/155), a decrease in platelet count (50.97%, 79/155), a decrease in albumin (92.26%, 143/155), an increase in lactate dehydrogenase (90.32%, 140/155), an increase in adenosine deaminase (88.39%, 137/155), and an increase in alanine aminotransferase (85.16%, 132/155), elevated aspartate aminotransferase (85.16%, 132/155), and elevated procalcitonin (52.90%, 82/155); 30 cases (19.35%) were positive for the Weil-Felix Test. There were 95 cases (61.29%) with abnormal chest imaging results, and 34 cases (21.94%) with abnormal abdominal ultrasound or CT results. Common complications were toxic hepatitis, pulmonary infection, organ failure, and acute kidney injury, etc. The misdiagnosis rate of the initial diagnosis of this disease was 75.48% (117/155). Doxycycline and symptomatic and supportive therapy were given, 154 patients (99.35%) were cured or improved and discharged from hospital.Conclusions:Tsutsugamushi disease is prevalent in summer and autumn in Guangdong Province. Before the onset, most of the patients have a history of field activities. Farmers are susceptible people. Its clinical manifestations are diverse, and can affect multiple systems and organs. There are many complications, and doxycycline can be used for anti-infection treatment, with a high cure rate.