ABSTRACT
Background& AimsThe Coronavirus Disease 2019 (COVID-19) has become a global epidemic and has caused a lasting and huge loss of life security, economic development and social stability in more than 180 countries around the world. Unfortunately, there is still no specific treatment for COVID-19 till now, therefore, at this point, all potential therapies need to be critically considered. LL-37 is one of the best-studied human antimicrobial peptide (AMPs) that has a broad-spectrum activity against bacteria and viruses. The use of living, genetically modified organisms (GMOs) is an effective approach for delivery of therapeutic proteins. The aim of this study was to determine the safety and efficacy of the Lactococcus lactis which has been genetically modified to produce the therapeutic human antimicrobial peptide LL-37 (herein after referred to cas001) in the patients of COVID-19. MethodsFirstly we constructed genetically modified food-grade probiotic, Lactococcus lactis, with sequence of seven tandem repeats of mature human LL-37 under control of the nisin-inducible nisA promoter to produce the cas001. A total of 20 healthy SD rats, half male and half female (There were five male and five female in the control group, the same in treatment group) were used to observe the acute toxic reaction and death after daily administration of cas001 for three weeks, which helps to provide necessary reference basis for clinical dose selection, verificaition of toxic reaction and possible target organs. According to the estimated clinical dosage of 1 x 108CFU /kg/day, considering the conversion of body surface area, the dose for rats should be multiplied by 6.17 to 6 x 108 CFU/kg/day. We administrated 100 times higher dose at 6 x 1010 CFU/ /kg/day to rats. In order to investigate the pharmacokinetics of cas001, male SD rats (body weight 250-300g, 1 x 1010 /animal, n=3) were given oral administration of LL-37 bacteria powder. The concentration of LL-37 in the blood before and after gavage was detected by ELISA kit (Hycult biotechnology Cat# HK321). Human clinical study was approved by Ethics committee of Chinese PLA General Hospital (S2020-074-04) and a total of 11 patients with mild symptoms were enrolled in Wuhan hankou hospital and Huoshenshan hospital. They were enrolled voluntarily and all patients signed informed consent. Among them, there were 5 males and 6 females, aged 55 {+/-} 12 (36-70) years old, and the duration from onset to medication enrollment was 35 {+/-} 19 (5-68) days. 6 patients were nucleic acid positive and 5 patients were nucleic acid negative when they were enrolled. All patients received the oral drug cas001 treatment according to requirement(1 x 109 CFU/capsule, 3 capsules/time, three times a day for 3weeks), with an average follow-up time of 33 {+/-} 15 days (see table 1 for the results). O_TBL View this table: org.highwire.dtl.DTLVardef@1845d29org.highwire.dtl.DTLVardef@1004b38org.highwire.dtl.DTLVardef@4a741aorg.highwire.dtl.DTLVardef@c73d5org.highwire.dtl.DTLVardef@188b563_HPS_FORMAT_FIGEXP M_TBL O_FLOATNOTable 1.C_FLOATNO O_TABLECAPTIONSerum biochemical detection in rats two weeks after gavage Serum biochemistry detection of rats at the end of two weeks (Vehicle[male] v.s. LL-37[male], n=5; Vehicle[female] v.s. LL-37[female], n=5). *represent p value<0.05. C_TABLECAPTION C_TBL FindingsWestern blot analysis shows that reasonable amount of LL-37 were induced by different concentrations of nisin, which means we have successfully constructed cas001. In the pre-clinical safety evaluation test, after three weeks administration of cas001, no adverse effects were observed on the rats body weight, food and water intake, hematological or serum biochemical parameters. The results showed that the LD50 of cas001 was higher than that of the 100 times of the expected clinical dose of 6 x 1010 CFU/day. These results showed that cas001 could be safe in animal experiments. In addition, rat pharmacokinetics results showed that the serum concentration of LL-37 reached peak 2 hours after gavage of cas001 and returned to basal level 6 hours after gavage. During study period, the volunteers did not feel any discomfort while taking the cas001 capsules, and two hours after oral administration, the concentration of LL-37 were increased in healthy volunteers. cas001 shows definite effect in the improvement of gastrointestinal symptoms and is possible to have effects in improving the systemic symptoms and respiratory symptoms and may play a role in the improvement of results of nucleic acid test and lung CT test. 11 patients enrolled showed good compliance, tolerance, subjective feeling and actively interacted with the doctors. None of the patients had any adverse reactions. ConclusionsBased on above observations, we conclude here that as an oral anti-viral agent, cas001 displayed good safety profiles. It is very hard to reach conclusion of clinical outcomes related to the cas001, although changes of several symptoms indicate encouraging findings.
ABSTRACT
BackgroundSARS-CoV-2-caused coronavirus disease (COVID-19) is posing a large casualty. The features of COVID-19 patients with and without pneumonia, SARS-CoV-2 transmissibility in asymptomatic carriers, and factors predicting disease progression remain unknown. MethodsWe collected information on clinical characteristics, exposure history, and laboratory examinations of all laboratory-confirmed COVID-19 patients admitted to PLA General Hospital. Cox regression analysis was applied to identify prognostic factors. The last follow-up was February 18, 2020. ResultsWe characterized 55 consecutive COVID-19 patients. The mean incubation was 8.42 (95% confidence interval [CI], 6.55-10.29) days. The mean SARS-CoV-2-positive duration from first positive test to conversion was 9.71 (95%CI, 8.21-11.22) days. COVID-19 course was approximately 2 weeks. Asymptomatic carriers might transmit SARS-CoV-2. Compared to patients without pneumonia, those with pneumonia were 15 years older and had a higher rate of hypertension, higher frequencies of having a fever and cough, and higher levels of interleukin-6 (14.61 vs. 8.06pg/mL, P=0.040), B lymphocyte proportion (13.0% vs.10.0%, P=0.024), low account (<190/{micro}L) of CD8+ T cells (33.3% vs. 0, P=0.019). Multivariate Cox regression analysis indicated that circulating interleukin-6 and lactate independently predicted COVID-19 progression, with a hazard ratio (95%CI) of 1.052 (1.000-1.107) and 1.082 (1.013-1.155), respectively. During disease course, T lymphocytes were generally lower, neutrophils higher, in pneumonia patients than in pneumonia-free patients. CD8+ lymphocytes did not increase at the 20th days after illness onset. ConclusionThe epidemiological features are important for COVID-19 prophylaxis. Circulating interleukin-6 and lactate are independent prognostic factors. CD8+ T cell exhaustion might be critical in the development of COVID-19.
ABSTRACT
Objective@#To investigate and analyze the epidemiological and clinical characteristics of some cases of coronavirus disease 2019 (COVID-19) in Beijing.@*Methods@#A retrospective study was conducted to analyze the data of 46 patients with COVID-19 in Beijing from 20th January 2020 to 8th February 2020 at the Fifth Medical Center of the PLA General Hospital. Features of clinical symptoms, laboratory inspections and imaging inspections were analyzed. Statistical analysis used Fisher exact test. If P<0.05, post-hoc test was used for pairwise comparison, and the statistics were corrected by Bonferroni test.@*Results@#Among the 46 patients included in this study, 27 were male and 19 were female. The age range was between 3 - 79 years old, and the mean age was (41.8 ± 16.3) years old. The average incubation period was (4.85 ± 3.00) days. A total of 26 cases (56.5%) were clustered patients, and 12 (26.1%), 23 (50.0%) and 11 patients (23.9%)were assigned to the mild group, common group, and sever group, respectively. Fever (39.8%), cough (27.6%), and fatigue (25.3%) was the main clinical symptom for these patients. The decrease in white blood cell counts occurred in 12 patients, four had the decrease in T lymphocyte counts, 17 had the decrease in CD4 + T lymphocyte counts, seven had the decrease in CD8 + T lymphocyte counts, 21 had the increase level of C-reactive protein (45.7%), and IL-6 level increased in 32 cases (69.6%), erythrocyte sedimentation rate increased in 20 cases(50.0%), serum ferritin level increased in 26 cases (56.5%), and blood lactate level increased in nine cases. There was a statistically significant difference in the number of cases in which the absolute value of T lymphocytes and of CD8 + T lymphocytes decreased among the mild, common and severe groups (all P<0.05). Comparing the number of cases in the three groups with elevated C-reactive protein, interleukin-6, erythrocyte sedimentation rate, serum ferritin and blood lactate levels, the differences were statistically significant (all P<0.05). The number of cases with elevated C-reactive protein levels was higher in severe group than that in mild and common group. The number of cases with elevated interleukin-6, erythrocyte sedimentation rate, and serum ferritin levels were higher in severe group than in mild group. The number of cases with elevated blood lactic acid levels was higher in severe group than in mild group. The differences between the above groups were statistically significant (both adjusted P<0.017). Analysis of chest X-rays showed that 34 patients (73.9%) had inflammation in the lungs.@*Conclusions@#The epidemiological characteristics of cases with COVID-19 in Beijing are mainly imported cases and clustered cases. The clinical manifestations are mainly fever , fatigue and cough. C-reactive protein, interleukin-6, red blood cell sedimentation rate, serum ferritin and blood lactate levels are higher in severe patients.
ABSTRACT
Objective:To investigate and analyze the epidemiological and clinical characteristics of 46 patients with corona virus disease 2019 (COVID-19) in Beijing City.Methods:A retrospective study was conducted to analyze the data of 46 patients with COVID-19 in Beijing from 20th January 2020 to 8th February 2020 at the Fifth Medical Center of the PLA General Hospital in Beijing City. Twelve, 23 and 11 patients were assigned to the mild group, common group and severe group, respectively. The epidemiological history, clinical characteristics, laboratory tests and imaging inspections were analyzed. Statistical analysis used Fisher exact test. If P<0.05, post- hoc test was used for pairwise comparison, and the statistics were corrected by Bonferroni test. Results:Among the 46 patients included in this study, 27 were male and 19 were female. The age range was between 3-79 years old, and the age was (41.8±16.3) years old. The average incubation period was (4.85±3.00) days. A total of 26 cases (56.5%) were clustered patients, and 26 cases had a history of staying in Wuhan, 10 cases had contact with Wuhan personnel. Fever (39 cases, 84.8%), cough (27 cases, 58.7%), and fatigue (25 cases, 54.3%) were the main clinical symptoms for these patients. The decrease in white blood cell counts occurred in 12 patients, four had the decrease in T lymphocyte percentage, 17 had the decrease in CD4 + T lymphocyte counts, seven had the decrease in CD8 + T lymphocyte counts, 21 had the increase level of C reactive protein (45.7%), and interleukin-6 (IL-6) level increased in 32 cases (69.6%), erythrocyte sedimentation rate (ESR) increased in 23 cases (50.0%), serum ferritin level increased in 26 cases (56.5%), and blood lactic acid level increased in nine cases. There were statistically significant differences in the proportion of cases with decreased absolute value of CD8 + T lymphocytes and T lymphocytes counts among the mild, common and severe groups (all P<0.05). Comparing the proportion of cases in the three groups with elevated C reactive protein, IL-6, ESR, serum ferritin and blood lactic acid levels, the differences were statistically significant (all P<0.05). The proportion of cases with elevated C reactive protein levels in severe group was higher than those in mild and common groups. The proportion of cases with elevated IL-6, ESR, and serum ferritin levels in severe and common group were higher than those in mild group. The proportion of cases with elevated blood lactic acid levels in severe group was higher than those in mild group. The differences between the above groups were statistically significant (all adjusted P<0.017). Analysis of chest X-rays results showed that 34 patients (73.9%) had inflammation in the lungs. Conclusions:The epidemiological characteristics of patients with COVID-19 in Beijing City are mainly imported cases and clustered cases. The clinical manifestations are mainly fever, fatigue and cough. C reactive protein, IL-6, ESR, serum ferritin and blood lactic acid levels are higher in severe patients.
ABSTRACT
Objective To compare and explore the differences of clinical characteristics between human adenovirus type 7 (HAdV7) and type 55 (HAdV55) infections in adults,and to provide evidences for clinical management.Methods The data of clinical manifestations,laboratory examinations,chest computed tomography and prognosis of 214 cases with HAdV7 and 235 cases with HAdV55 infections from 2012 to 2018 were retrospectively collected and analyzed.The chi-square test was used for the categorical variables,and the rank sum test was used for the continuous variables of non-normal distribution.Results Compared with patients in HAdV55,those in HAdV7 group displayed more diarrhea (12.1% [26/214] vs 2.6% [6/235],x2 =15.583),more laryngeal lymphatic follicles (33.2% [71/214] vs 17.9% [42/235],x2 =23.566),more tonsil enlargement (56.5% [121/214] vs 20.0% [47/235],x2 =63.870) with secretions (33.2% [71/214] vs 11.5% [27/235],x2 =30.878),more leukocytosis (24.8% [53/214] vs 14.0% [33/235],x2 =8.318),more monocytosis (78.0% [167/214] vs 52.8% [124/235],x2 =31.364),more thrombocytosis (7.1% [15/212] vs 3.8% [9/235],x2 =5.835),more elevated level of C-reactive protein (80.8% [97/120] vs 64.3% [137/213],x2 =10.020),more abnormalities of liver function (alamne aminotransferase [ALT]:13.5% [28/207] vs 5.6% [13/232],x2 =8.111) and myocardial enzymes (creatine kinase [CK]:37.6% [77/205] vs 26.4% [61/231],x2 =6.246),creatine kinase isoenzymes ([CK-MB]:35.6% [73/205] vs 11.1% [24/216],x2 =35.600),and all the differences were statistically significant (all P < 0.05,0.01).Compared with patients in HAdV7 group,those in HAdV55 group displayed longer duration of diseases (7 [5]days vs 6[5] days,Z=-2.632),more sore throat (72.8% [171/235] vs 62.1% [133/214],x2 =0.016),more headache (62.1% [146/235] vs 16.4% [35/214],x2 =97.527),more pharyngeal congestion (93.6% [220/235]vs 74.8% [160/214],x2 =30.602),more leukopenia (14.0% [33/235] vs 24.8% [53/214],x2 =8.318),and more elevated level of lactate dehydrogenase (12.6% [29/230] vs 6.3% [13/205],x2 =4.881),more pneumonia (71.5% [168/235] vs31.8% [68/214],x2 =70.846) (all P<0.05,0.01).Conclusions The clinical characteristics of HAdV7 and HAdV55 infections in adults are different.The type 55 infection is more likely to develop to pneumonia,while the type 7 infection has wider tissue tropism.
ABSTRACT
Objective@#o investigate the features of pathogenic bacteria for community-acquired bloodstream infection due to Gram-negative bacilli in patients with liver cirrhosis and optimal therapeutic strategy.@*Methods@#A retrospective analysis was performed for the clinical data of patients with liver cirrhosis who were admitted to 302 Hospital of PLA due to community-acquired bloodstream infection from January 2010 to December 2015, and a statistical analysis was performed for their clinical features, pathogenic bacteria, and results of drug sensitivity test. The Pearson chi-square test was used for comparison of rates, and the Wilcoxon rank sum test was used for comparison of ranked data.@*Results@#A total of 240 patients (including 178 male patients) with liver cirrhosis caused by various reasons were enrolled, with a mean age of 51.7 ± 11.1 years, an overall clinical remission rate of 80.42%, and an ineffective/mortality rate of 19.58%. The patients who used sensitive antibiotics within 12 hours after the onset of community-acquired bloodstream infection achieved a significantly higher improvement rate than those who used such drugs at more than 12 hours after onset (88.2% vs 58.1%, P < 0.001). The improvement rate achieved by the application of sensitive antibiotics at more than 12 hours after onset decreased with the increase in the Child-Pugh grade (P < 0.05). A total of 245 strains of Gram-negative bacilli were isolated, among which the six most common ones were 135 strains of Escherichia coli (55.1%), 62 strains of Klebsiella pneumoniae (25.3%), 16 strains of Aeromonas (6.5%), 4 strains of non-typhoidal Salmonella (1.6%), 3 strains of Enterobacter cloacae (1.2%), and 2 strains of Acinetobacter baumannii (0.8%). These Gram-negative bacilli had the highest sensitivity to meropenem (98.5%), followed by imipenem (97.9%), amikacin (97.5%), piperacillin/tazobactam (94.7%), cefmetazole (93.7%), and cefoperazone/sulbactam (93%). Different bacteria had different sensitivities to antibiotics.@*Conclusion@#Once community-acquired bloodstream infection occurs in patients with liver cirrhosis, highly sensitive antibiotics should be used as early as possible. Cefoperazone/sulbactam, piperacillin/tazobactam, imipenem, and meropenem can be used as first-line empirical antibiotics, and drug combination should be considered when necessary.
ABSTRACT
Objective To investigate the distribution and the rate of drug sensitivity of dysenteric bacilli in Beijing from 2003 to 2004 so as to guide the choice of antibacterial. Methods This was a retrospective study. 254 shigellae were cultured from stool. The group and serotype were identified with biochemical and serologic tests. Kirby-Bauer method was used to detect the drug sensitivity rate. Results There were 196 strains of Shigellae flexneri and 58 Shigellae sonnei a-mong 254 strains of shigellae. The number of F4 was 106 strains in Shigellae flexneri. SMZ resistance rate was the highest ; fluoroquinolones, the third generation cephalosporins and phosphonomycins had much more sensitivity. The drug sensitivity rate to fluoroquinolones between Shigellae flexneri and Shigellae sonnei was different. Conclusion In 2003 and 2004, Shigellae flexneri is the dominant group in 254 Shigellae in Beijing. F4 is the dominant serotype among Shigellae flexneri. On antibacterial effects,fluoroquinolones is one of the most effective drug, but had a certain resistance rate. If the pathogen is resistant to fluoroquinolones, the third generation cephalosporins or phosphonomycins can be used.
ABSTRACT
AIM:To construct hybrid nucleic acid vaccines which contain Plasmodium falciparum merozoite surface protein 1 (MSP1 ) block 1 7gene and gene fragment coding for several T cell epitopes. METHODS:MSP1 block 1 7gene of FUP strain was connected with a polyvalent gene fragment which contains several T cell epitopes from MSA1 ,MSA2 ,RESA,IL- 1 and Tetanus toxin,the connected gene fragment (hybrid gene,HG) was cloned into eukaryotic expression plasmid p CDNA3.1 (- ) ,VR1 0 1 2 for intracellular expression,VR1 0 1 2 /TPA for secretion,the recombinants were identified by PCR and restriction enzyme digestion. RE- SUL TS &CONCLUSION:The hybrid nucieic acid vaccines:VR1 0 1 2 /HG,VR1 0 1 2 /TPA/ HG and p CDNA3.1 /HG were successfully constructed.