The Protective Effects of the A/ZJU01/ PR8/2013 Split H7N9 Avian Influenza Vaccine Against Highly Pathogenic H7N9 in BALB/c Mice.

BACKGROUND/AIMS: Since the first case of novel H7N9 infection was reported, China has experienced five epidemics of H7N9. During the fifth wave, a highly pathogenic H7N9 strain emerged. In order to assess whether the H7N9 vaccine based on A/Zhejiang/DTID-ZJU01/2013(H7N9) was effective in protecting against highly pathogenic H7N9, we conducted this study. METHODS: Groups of mice were immunized twice by intraperitoneal injection with 500 µl of either split vaccine alone or MF59-adjuvanted vaccine. Serum was collected 2 weeks after the second vaccine booster. The hemagglutinin inhibition test was conducted on vaccine seed and highly pathogenic H7N9 to evaluate the neutralization of highly pathogenic H7N9. We also immunized mice and challenged them with highly pathogenic H7N9. Mice were observed for illness, weight loss, and death at 1 week and 2 weeks post-infection. Then, the mice were sacrificed and lungs were removed. Antibody responses were assessed and pathological changes in the lung tissue were evaluated. RESULTS: The ability of serum to neutralize highly pathogenic H7N9 was reduced. In mice, highly pathogenic H7N9 was more virulent than A/Zhejiang/DTID-ZJU01/2013(H7N9). After challenge with highly pathogenic H7N9, all mice died while mice challenged with A/Zhejiang/DTID-ZJU01/2013(H7N9) all recovered. The A/ZJU01/PR8/2013 split H7N9 avian influenza vaccine was able to protect against infection with highly pathogenic H7N9 in mice, with or without MF59. Moreover, H7N9 vaccine adjuvanted with MF59 produced high antibody levels, which lead to better protection. CONCLUSIONS: The A/ZJU01/PR8/2013 split H7N9 avian influenza vaccine based on A/Zhejiang/DTID-ZJU01/2013(H7N9) is effective in protecting against highly pathogenic H7N9. H7N9 vaccine adjuvanted with MF59 offers better protection against infection with highly pathogenic H7N9. In order to make the H7N9 vaccine applicable to humans, further clinical trials are required to evaluate MF59 adjuvanted vaccine. Meanwhile, the vaccine strain should be updated based on the highly pathogenic H7N9 gene sequence.

[Preparation and quality evaluation of tanshinone IIA microemulsion for parenteral injection].

OBJECTIVE: To study the prescription and preparation technology of tanshinone IIA microemulsion for parenteral injection, and to evaluate its quality. METHODS: The prescription was selected and optimized through single-factor test, compatibility experiment and the pseudo-ternary phase diagram method. The preparation technology was investigated, and the droplet morphous, particle diameter, zeta potential, stability and haemolyticus were evaluated. RESULTS: The prescription composition of tanshinone IIA microemulsion was MCT:Solutol HS-15: fabaceous lecithin: absolute alcohol = 9:10:5:6(m/m), oil phase: aqueous phase = 1:10, with the drug-loaded of 1. 0 mg/g. The acquired microemulsion exhibited salmon pink,uniform and transparent, with the average particle diameter of 16. 04 nm, Zeta potential of -11. 57 mV, and the encapsulation efficiency of 98. 53%. The stability result showed that tanshinone IIA content in microemulsion was influenced by high temperature and illumination, indicating tanshinone IIA microemulsion should to be stored at low temperature and protected from light. The preparation was without hemolytic crisis. CONCLUSION: The preparation of tanshinone IIA micro- emulsion is simple,corresponding to the main index of parenteral injection and offering the basis for new dosage form development of tanshinone IIA.

Clinical and pathogen features of COVID-19-associated infections during an Omicron strain outbreak in Guangzhou, China.

Although the Omicron variant has been associated with greater transmissibility and tropism of the upper respiratory tract, the clinical and pathogenic features of patients infected with the Omicron variant during an outbreak in China have been unclear. Adults with COVID-19 were retrospectively enrolled from seven medical centers in Guangzhou, China, and clinical information and specimens ( BALF, sputum, and throat swabs) from participants were collected. Conventional detection methods, metagenomics next-generation sequencing (mNGS), and other methods were used to detect pathogens in lower respiratory tract samples. From December 2022 to January 2023, we enrolled 836 patients with COVID-19, among which 56.7% patients had severe/critical illness. About 91.4% of patients were infected with the Omicron strain (BA.5.2). The detection rate of possible co-infection pathogens was 53.4% by mNGS, including Klebsiella pneumoniae (16.3%), Aspergillus fumigatus (12.2%), and Pseudomonas aeruginosa (11.8%). The co-infection rate was 19.5%, with common pathogens being Streptococcus pneumoniae (11.5%), Haemophilus influenzae (9.2%), and Adenovirus (6.9%). The superinfection rate was 75.4%, with common pathogens such as Klebsiella pneumoniae (26.1%) and Pseudomonas aeruginosa (19.4%). Klebsiella pneumoniae (27.1%% vs 6.1%, P < 0.001), Aspergillus fumigatus (19.6% vs 5.3%, P = 0.001), Acinetobacter baumannii (18.7% vs 4.4%, P = 0.001), Pseudomonas aeruginosa (16.8% vs 7.0%, P = 0.024), Staphylococcus aureus (14.0% vs 5.3%, P = 0.027), and Streptococcus pneumoniae (0.9% vs 10.5%, P = 0.002) were more common in severe cases. Co-infection and superinfection of bacteria and fungi are common in patients with severe pneumonia associated with Omicron variant infection. Sequencing methods may aid in the diagnosis and differential diagnosis of pathogens. IMPORTANCE: Our study has analyzed the clinical characteristics and pathogen spectrum of the lower respiratory tract associated with co-infection or superinfection in Guangzhou during the outbreak of the Omicron strain, particularly after the relaxation of the epidemic prevention and control strategy in China. This study will likely prompt further research into the specific issue, which will benefit clinical practice.

Effect of Recombinant Human Granulocyte Colony-Stimulating Factor for Patients With Coronavirus Disease 2019 (COVID-19) and Lymphopenia: A Randomized Clinical Trial.

Importance: Lymphopenia is common and correlates with poor clinical outcomes in patients with coronavirus disease 2019 (COVID-19). Objective: To determine whether a therapy that increases peripheral blood leukocyte and lymphocyte cell counts leads to clinical improvement in patients with COVID-19. Design, Setting and Participants: Between February 18 and April 10, 2020, we conducted an open-label, multicenter, randomized clinical trial at 3 participating centers in China. The main eligibility criteria were pneumonia, a blood lymphocyte cell count of 800 per µL (to convert to ×109/L, multiply by 0.001) or lower, and no comorbidities. Severe acute respiratory syndrome coronavirus 2 infection was confirmed with reverse-transcription polymerase chain reaction testing. Exposures: Usual care alone, or usual care plus 3 doses of recombinant human granulocyte colony-stimulating factor (rhG-CSF, 5 µg/kg, subcutaneously at days 0-2). Main Outcomes and Measures: The primary end point was the time from randomization to improvement of at least 1 point on a 7-category disease severity score. Results: Of 200 participants, 112 (56%) were men and the median (interquartile range [IQR]) age was 45 (40-55) years. There was random assignment of 100 patients (50%) to the rhG-CSF group and 100 (50%) to the usual care group. Time to clinical improvement was similar between groups (rhG-CSF group median of 12 days (IQR, 10-16 days) vs usual care group median of 13 days (IQR, 11-17 days); hazard ratio, 1.28; 95% CI, 0.95-1.71; P = .06). For secondary end points, the proportion of patients progressing to acute respiratory distress syndrome, sepsis, or septic shock was lower in the rhG-CSF group (rhG-CSF group, 2% vs usual care group, 15%; difference, -13%; 95%CI, -21.4% to -5.4%). At 21 days, 2 patients (2%) had died in the rhG-CSF group compared with 10 patients (10%) in the usual care group (hazard ratio, 0.19; 95%CI, 0.04-0.88). At day 5, the lymphocyte cell count was higher in the rhG-CSF group (rhG-CSF group median of 1050/µL vs usual care group median of 620/µL; Hodges-Lehmann estimate of the difference in medians, 440; 95% CI, 380-490). Serious adverse events, such as sepsis or septic shock, respiratory failure, and acute respiratory distress syndrome, occurred in 29 patients (14.5%) in the rhG-CSF group and 42 patients (21%) in the usual care group. Conclusion and Relevance: In preliminary findings from a randomized clinical trial, rhG-CSF treatment for patients with COVID-19 with lymphopenia but no comorbidities did not accelerate clinical improvement, but the number of patients developing critical illness or dying may have been reduced. Larger studies that include a broader range of patients with COVID-19 should be conducted. Trial Registration: Chinese Clinical Trial Registry: ChiCTR2000030007.

[Analysis of acute physiology and chronic health evaluation III in the 54 patients with severe acute respiratory syndrome].

OBJECTIVE: To evaluate the acute physiology and chronic health evaluation III (APACHE III) in the severe acute respiratory syndrome (SARS) patients and its significance in prognosis. METHODS: The clinical data of 54 SARS patients, including survivors (43 cases) and nonsurvivors (11 cases) were collected and evaluated with APACHE III scoring system. The correlation of scores and prognosis was evaluated. RESULTS: The scores of the nonsurvivors were higher remarkably than those of the surviving group(P<00.01). The scores of the acute physiology score (APS) and year score (YS) in the death group were higher obviously than those of the surviving group(both P<0.01). Elderly patients with severe disease had a high mortality. The scores of APACHE III had positive correlation with the over all fatality rate. When the scores of APACHE III was higher than 60, the fatality rate increased obviously. CONCLUSION: The scores of APACHE III in the SARS are correlated with the patient's condition and prognosis.

[Relationship between psychological distress and T lymphocyte in HIV/AIDS patients].

OBJECTIVE: To explore the relationship between psychological distress and T lymphocyte counts in HIV/AIDS patients. METHODS: A total of 102 HIV/AIDS patients were measured by symptom check list (SCL-90), self-rating depressive scale (SDS) and self-rating anxiety scale (SAS). Patients were divided into 2 groups based on CD4+ T lymphocyte counts < 0.2 x 10(9)/L (group A) and > or = 0.2 x 10(9)/L(group B). RESULTS: 77 cases (75.49%) had psychological problems, including depression, relationship problems, psychosis, force etc. The prevalence of depression and anxiety were 67.65% (69/102) and 43.13% (44/102) respectively. The symptom of depression and anxiety of patients in group A were severer than those in group B (P < 0.05). The CD4+ T lymphocyte counts were significantly negatively correlated with the total score, depression score, paranoid score and psychosis score of SCL-90 (all P <0.05). CONCLUSION: Most of the HIV/AIDS patients were in an obviously abnormal psychological status. The psychological distress symptom of HIV/AIDS patients might had negative effects on the number of CD4+ T lymphocyte.

[Dose of glucocorticosteroids in the treatment of severe acute respiratory syndrome].

OBJECTIVE: To survey the dose of glucocorticosteroids administered in patients with severe acute respiratory syndrome (SARS) and assess the effect of glucocorticosteroid doses in improving the patients' lung function. METHODS: A retrospective analysis was conducted among 225 SARS patients treated in our in 2003. Oxygenation index was used as the effectness index, and the criteria for effectiveness was defiend as increase of the value of OI by 20% or above. RESULTS: Glococoticostecoids were used in 59.56% of the SARS cases. The average value of OI before intravenous use of glucocorticosteroids was 237.08 mmHg, and that after the administration was 335.08 mmHg. The glucocorticosteroid doses that produce better effects were 1-3 mg/kg and 160-240 mg daily, with the total accumulative dose of 1000-2000 mg. The optimal duration of glucocorticosteroid use was 8-14 days. CONCLUSIONS: For SARS treatment, Glucocorticosteroids can effectively ameliorate the SARS patients' lung symptoms and improve the lung function. The appropriate daily dose of glucocorticosteroids is 1-3 mg/kg or 160-240 mg/d for a duration of 8-14 d; the accumulative dose should be controlled around 1500 mg.

Tumor necrosis factor--alpha gene promoter polymorphisms in Chinese patients with nonalcoholic fatty liver diseases.

BACKGROUND: Nonalcoholic Fatty Liver Disease (NAFLD) encompasses a histopathological spectrum of clinical conditions such as simple fatty liver (steatosis), nonalcoholic steatohepatitis (NASH), and a variant that has degrees of fibrosis. Tumor Necrosis Factor-alpha (TNF-alphalpha) is considered essential for NAFLD. Therefore, we investigated the correlation between TNF-alphalpha gene promoter polymorphism and NAFLD in this human study. PATIENTS AND METHODS: The TNF-alpha gene polymorphisms at position -238 and -308 were analyzed in 189 Chinese patients with NAFLD and 138 healthy controls by using polymerase chain reaction and restriction fragment length polymorphism assay. The serum levels of TNF-alpha in both patient and control groups were measured by ELISA. The associations of TNF-alpha polymorphism and serum TNF-alpha, and/or insulin resistance, and/or clinical features were analyzed. RESULTS: The carrier frequencies of TNF-alpha gene polymorphism with G/A mutation at -238 were significantly higher in the patients with NAFLD than those in the control subjects (p < 0.05). However, there were no significant differences between the NAFLD patients and control subjects in the polymorphisms at -308 (p > 0.05). In addition, the serum level of TNF-alpha was markedly higher in the patients with NAFLD than in the control subjects (p < 0.05). There were significant associations between TNF-alpha gene polymorphism in the -238 A allele and increased serum TNF-alpha, insulin resistance, as well as increased body mass index in the NAFLD patients. CONCLUSIONS: The results indicate that the TNF-alpha gene polymorphism at position -238 is associated with susceptibility of nonalcoholic Fatty Liver Disease in a Chinese population.

[Association of serum leptin and insulin resistance with nonalcoholic fatty liver disease].

OBJECTIVE: To investigate the relationship among serum leptin, homeostasis model assessment of insulin resistance (HOMA-IR), body mass index (BMI) and dyslipidema in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: Eighty-two patients with NAFLD were divided into mild, moderate and severe NAFLD groups according histological examination results of the liver. Twenty healthy volunteers were chosen as the normal control (NC) group. Fasting insulin, glucose, leptin and lipid levels were measured in the 82 patients with NAFLD and the BMI calculated. IR index of the patients was calculated according to the HOMA method. RESULTS: Leptin, HOMA-IR index, BMI and dyslipidemia showed significant differences between NAFLD and NC groups (P<0.05 or 0.01). Leptin and HOMA-IR index increased with the exacerbation of NAFLD, both of which were positively correlated with the severity of NAFLD. CONCLUSION: Increased leptin level, HOMA-IR, BMI and dyslipidemia can be important risk factors of NAFLD, and serum leptin level and HOMA-IR are positively correlated with the severity of NAFLD.