Objective To analyze the clinical features of patients with coronavirus disease 2019 (COVID-19) in Shanghai and to investigate the risk factors for disease progression to severe cases. Methods The clinical data of 292 adult patients with COVID-19 hospitalized in Shanghai Public Health Clinical Center from January 20, 2020 to February 10, 2020 were retrospectively analyzed, including 21 severe patients and 271 mild patients. The demographic characteristics, epidemiological history, history of underlying diseases and laboratory examinations were compared between the two groups. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using hi-square test. The binary logistic regression equation was used to analyze the risk factors for the progression of patients to severe cases. Results Among the 292 patients, 21 were severe cases with the rate of 7.2% (21/292). One patient died, and the mortality rate was 4.8% in severe patients. The severe patients aged (65.0±15.7) years old, 19 (90.5%) were male, 11 (52.4%) had underlying diseases, 7 (33.3%) had close relatives diagnosed with COVID-19. The mild patients aged (48.7±15.7) years old, 135 (49.8%) were male, 74 (27.3%) had underlying diseases, 36 (13.3%) had close relatives diagnosed with COVID-19. The differences between two groups were all significant statistically ( t =-4.730, χ 2 =12.930, 5.938 and 4.744, respectively, all P <0.05). Compared with the mild patients, the levels of absolute numbers of neutrophils, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, creatinine, serum cystatin C, C reactive protein (CRP), procalcitonin , D -dimer, pro-B-type natriuretic peptide (proBNP), serum myoglobin, creatine kinase (CK), creatine kinase isoenzyme (CK-MB), serum troponin I (cTnI) in severe patients were all significantly higher ( U =2 091.5, 1 928.0, 1 215.5, 729.0, 1 580.5, 1 375.5, 917.5, 789.5, 1 209.0, 1 434.0, 638.0, 964.5, 1 258.0 and 1 747.5, respectively, all P <0.05), while the levels of lymphocyte count, albumin, transferrin, CD3 + T lymphocyte count, CD8 + T lymphocyte count and CD4 + T lymphocyte count in severe patients were all significantly lower ( U =1 263.5, t =4.716, U =1 214.0, 962.0, 1 167.5 and 988.0, respectively, all P <0.05). Further logistic regression analysis showed that the albumin (odds ratio ( OR )=0.806, 95% CI 0.675-0.961), CRP ( OR =1.016, 95% CI 1.000-1.032), serum myoglobin ( OR =1.010, 95% CI 1.004-1.016), CD3 + T lymphocyte count ( OR =0.996, 95% CI 0.991-1.000) and CD8 + T lymphocyte count ( OR =1.006, 95% CI 1.001-1.010) at admission were independent risk factors for the progression of COVID-19 patients to severe illness (all P <0.05). Conclusions Severe cases of patients with COVID-19 in Shanghai are predominantly elderly men with underlying diseases. Albumin, CRP, serum myoglobin, CD3 + T lymphocyte count and CD8 + T lymphocyte count could be used as early warning indicators for severe cases, which deserve more clinical attention.

Wuhan is the city with the most serious outbreak of corona virus disease 2019 (COVID-19) in China. The outbreak of community has exhausted the current medical resources. With integrating local and support medical resources from other province, Wuhan City has rapidly rebuilt a new emergency medical system of classified treatment, and effectively responded to the overload medical demand after the outbreak in the community.

Objective: To evaluate the value of T cell spot test of tuberculosis infection(T-SPOT.TB) and inflammatory indicators for diagnosis of active tuberculosis in patients with fever of unknown origin (FUO). Methods: Patients with FUO in Tongji Hospital from Jan 1st 2014 to Feb 28th 2015 were retrospectively enrolled, and general condition, laboratory examination including T-SPOT.TB, blood routine test, procalcitonin (PCT), high sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR), lactate dehydrogenase (LDH), serum ferritin (SF) and final diagnosis were collected and analyzed. Results: A total of 395 hospitalized patients with FUO were retrospectively enrolled into this study, among which there were 36 (9.11%) confirmed active tuberculosis (including 7 pulmonary cases and 29 extra-pulmonary cases), 189 (47.85%) bacterial infections, 50 (12.66%) viral infections, 4 (6.32%) fungal infections, 20 (5.06%) neoplastic diseases, 51(12.91%) autoimmune diseases, 25 (6.32%) other diseases. While 20 (5.06%) patients remained un-diagnosed. The sensitivity of T-SPOT.TB for the diagnosis of active TB in patients with FUO was 80.56% (95%CI: 63.43%-91.20%), and the specificity was 83.57% (95%CI: 79.23%-87.16%). The positive predictive value was 32.95% (95%CI: 23.52%-43.89%), and the negative predictive value was 97.72% (95%CI: 95.16%-99.00%). There were significant differences in positive LDH levels (187[141, 255] U/L vs 209[160, 343] U/L) and SF levels (296.2[191.3, 494.8] g/L vs 528.1[281.1, 1 022.0] μg/L) between active tuberculosis group and bacterial infection group (χ²=77.692, H=13.442, H=16.142, all P<0.05). The combination of T-SPOT.TB and multiple inflammatory indicators obtained most valuable efficiency (AUC=0.866) for TB diagnosis. Similarly, there were significant differences in positive ESR (31[15, 78] mm/1 h vs 10[6, 19] mm/1 h), ratio of neutrophil granulocytes ([71.17±12.59]% vs [57.08±20.38]%) between active tuberculosis group and viral infection group (H=32.797, F=6.171, all P<0.05). The combination acquired most valuable efficiency (AUC=0.929). Conclusions For patients with FUO, T-SPOT.TB combined with inflammatory indicators are valuable for the diagnosis of active tuberculosis.

Objective: To determine the clinical significance of nontuberculous mycobacteria (NTM) isolated from respiratory specimens. Methods: Clinical data of patients with NTM strains isolated from the respiratory tract between January 2014 and February 2017 were retrospectively analyzed. Clinical significance of NTM isolated strains was evaluated based on diagnostic criteria of NTM pulmonary diseases from American Thoracic Society (ATS). Quantitative data of two groups were analyzed by independent t test. Categorical data were analyzed by Pearson χ² test or Fisher exact test. Results: Totally 352 NTM strains from 257 patients had been isolated between January 2014 and February 2017. Among 15 identified NTM species, M. intracellular (51.7%, 182/352), M. abscessus (25.6%, 90/352), and M. avium (10.8%, 38/352) were predominant. Of the 157 patients with full clinical data involved in the analysis, 58 (36.9%) patients were determined to have definite disease, and 34 patients (21.7%) were designated as probable disease candidates, and 16 (10.2%) patients were regarded as uncertain disease, and 49(31.2%) patients were diagnosed as unlikely disease. The age of 58 patients with definite disease was (63.9±12.7) years, and 48.3% (28/58) were female. M. intracellulare (55.2%, 32/58) was the main cause of pulmonary NTM disease, followed by M. abscessus (25.9%, 15/58) and M. avium (12.1%, 7/58), while other NTM species only accounted for 6.8% (4/58). Definite cases with M. intracellulare, M. abscessus, M. avium, M. kansasii, M. marseillense, and M. columbia accounted for 35.2% (32/91), 57.7% (15/26), 7/15, 2/3, 1/3 and 1/2, respectively, among patients with corresponding isolations, while patients with other species of isolation did not meet the diagnostic criteria. Patients with clinical significant isolation of NTM were older than those without clinical significance (χ²=3.603, P=0.000), and proportion of anti-acid staining positivity of patients with clinical significance was higher than that of patients without clinical significance (χ²=18.815, P=0.000). The proportion of M. abscessus in patients with clinical significance was higher than that in patients without clinical significance (χ²=6.313, P=0.012). However, there was no significant difference between the two groups in the isolation of M. Gordon (Fisher exact test, P=0.028). The proportion of M. abscessus lung disease in women was 11/15, which was higher than that of M. intracellulare lung disease (41.5%, 17/41), and the difference between the two groups was statistically significant (χ²=4.462, P=0.035). There was no significant difference of clinical symptoms and underlying diseases in NTM lung disease among different groups (all P>0.05). In these patients with definite disease, 39.7% (23/58) of them manifested the upper lobe cavitary form, 53.4% (31/58) exhibited nodular bronchiectatic form, and only 6.9% (4/58) exhibited unclassified form. The upper lobe cavitary form (43.8%, 14/32) and the nodular bronchiectatic form (53.1%, 17/32) dominated in patients with M. intracellulare lung disease. M. abscessus lung disease was dominated by the nodular bronchiectatic form (11/15) while the upper lobe cavitary form only accounts for 3/15. There was no significant difference of image characteristics between NTM lung disease and other different groups (all P>0.05). Conclusions 36.9% of the patients with NTM isolates met the ATS diagnostic criteria for NTM lung disease. Different species have different clinical significance. M. intracellulare and M. abscessus are the most predominant NTM isolated species that cause NTM lung disease. Majority of patients manifest as the upper lobe cavitary form, followed by the nodular bronchiectatic form.

Objective: To analyze and summarize the clinical characteristics of tubercular lymphadenitis, and to improve the ability of diagnosis. Methods: Clinical records of 129 patients first confirmed with tubercular lymphadenitis were collected retrospectively from Nanfang Hospital of Southern Medical University between January 2012 and December 2016. The categorical variables were described with the percentage (%) and compared with the chi-squaue test. Non-normal distribution data were described with M(P₂₅, P₇₅) and compared with rank sum test. Results: The disease courses were different in all cases, mostly of 1-3 months (45.7%). Among the cases, 83 cases (73.6%) complained of lymph node enlargement. The predominant involved lymph node site was cervical (56.6%) with main presentation of single lymph node (61.2%). Only a few cases presented with fever (34.1%). The positive rate of histological examinations was 94.3%, while the positive rate of T cell spot test of tuberculosis infection (T-SPOT.TB) test was 93.3% and purified protein derivative (PPD) test was 69.6%. In the diagnosis of tubercular lymphadenitis, 100 cases (77.5%) were confirmed by histological examinations, 27 cases (20.9%) were given diagnostic treatment, and only 2 case (1.6%) was confirmed by culture. The average period of diagnosis was (10.4±6.5) days. The median age of patients with fever was 50.5 years old with a median disease course of 2.5 months, while the median age of patients fever was 35(24, 49) years old with a median disease course of 1.2(0.5, 6.0) months. The differences between two groups were statistically significant (Z=-3.118 and -2.982, respectively, both P<0.05). Patients with fever had higher proportion of swollen deep lymph nodes (54.5% vs 11.8%), elevated white blood cell counts (34.1% vs 7.1%) and neutrophils (31.8% vs 1.8%), elevated erythrocyte sedimentation rate (97.1% vs 56.1%), elevated C-reactive protein (95.0% vs 40.0%) and received diagnostic treatment (47.7% vs 7.1%) than patients with no fever (χ²=27.337, 15.545, 13.567, 19.347, 25.410 and 28.974, respectively, all P<0.05). Conclusions Most patients of tubercular lymphadenitis do not present with typical symptoms which might lead to misdiagnose in early stage. The histological examinations and T-SPOT.TB test are especially essential, and histological examinations is the most important diagnostic method. Patients without symptoms of tuberculous poisoning are more common in young people, and the confirmation of diagnosis are mainly based on histological examinations. Patients with symptoms of tuberculous poisoning are more common in middle-aged, with longer duration and deep lymph node involved, which is more serious and nearly half of which are confirmed with diagnostic treatment.

Objective: To investigate the epidemiologic features of interhospital, intrahospital, interdepartmental and intradepartmental infections of carbapenem-resistant Acinetobacter baumannii (CRAB) isolated from clinical specimens. Methods: From September 2015 to September 2016, 540 non-repetitive strains of Acinetobacter baumannii were clinically isolated from 38 tertiary hospitals in Anhui Province, of which 460 strains were confirmed by antimicrobial resistance surveillance in Anhui Province. Agar plate dilution method was used to determine the MIC of 16 antibiotics and to select 312 CRAB isolates for drug sensitivity test. Pulsed field gel electrophoresis (PFGE) was used to analyze homology of 145 strains. PCR and clonal sequencing were used to determine the corresponding genotypes. Results: All the 312 CRAB strains were multiple antibiotic resistant strains. The drug resistance rates to imipenem and meropenem were 67.9% and 70.6%, respectively. All the 145 strains were divided into 80 different types (PT1-PT80) by PFGE. Only one strain was found in the 60 types and two or more strains were found in the other 20 types. Genotype bla_OXA-51 was detected in 145 strains (100%), genotype bla_OXA-23 was detected in 134 of strains (92.41%), genotype bla_OXA-24 was detected in 1 strain (0.69%), and subtype of bla_IMP-4 was detected in 4 strains (2.76%). Genotype bla_OXA-58, bla_VIM, bla_SIM, bla_NDM and bla_KPC were undetectable. Conclusions Homologous analysis indicates that all 145 strains of CRAB exhibit the interhospital, intrahospital, interdepartmental and intradepartmental transmission. Genotype bla_OXA-51 is the inherent drug resistant associated gene in the chromosome of AB and genotype bla_OXA-23 is the most common carbapenemase gene in Anhui Province and the main genotypes of resistance transmission.

Objective: To explore the changes of the peripheral invariant natural killer T (iNKT) cells in patients with human immunodeficiency virus (HIV) infection. Methods: A total of 101 patients with HIV infection including 52 asymptomatic patients and 49 acquired immunodeficiency syndrome (AIDS) patients were enrolled in the study from June 2016 to July 2017. Flow cytometry was used to detect iNKT cells, CD4⁺ T cells and CD8⁺ T cells, and the relationship among them and HIV RNA was studied. At same time, 12 healthy persons were enrolled as control group. T test or variance analysis, rank sum test, Chi-square test and Fisher exact test were used for statistical analysis. Results: In HIV infected asymptomatic patients, AIDS patients and healthy controls, iNKT cells were 0.135% (0.066%, 0.228%), 0.058% (0.034%, 0.100%) and 0.385% (0.205%, 0.600%), respectively, and the difference was statistical significant (Z=40.113, P<0.01). CD4⁺ T cell counts in the three groups were (340.82±119.26) cells/μL, (72.73±61.84) cells/μL and (555.17±229.43) cells/μL, respectively, and the difference was statistical significant (t=113.79, P<0.01); CD8⁺ T cell counts in the three groups were (842.29±423.68) cells/μL, (540.43±257.85) cells/μL and (875.92±516.45) cells/μL, respectively, and the difference was statistical significant (t=9.423, P<0.01). Ratios of CD4⁺ /CD8⁺ T cells in the three groups were 0.490 (0.240, 0.695), 0.120 (0.030, 0.210) and 0.600 (0.475, 0.895), respectively, and the difference was statistical significant (Z=53.603, P<0.01). iNKT cell counts in patients with or without hepatitis B virus infection, pneumocystis pneumonia, oral mold infection, treponema pallidum, latent tuberculosis or EB virus infection were not significantly different (Z=0.244, 2.325, 2.393, 0.168, 1.183 and 0.454, respectively, all P>0.05). There were correlations between iNKT cells and CD4⁺ T cells, CD4⁺ /CD8⁺ T cells (r=0.513 and 0.261, respectively, both P<0.05), and no relationship was found between iNKT cells and CD8⁺ T cells (r=0.155, P=0.126). In HIV infected asymptomatic patients and AIDS patients, iNKT cells was not associated with HIV RNA (r=-0.113 and -0.111, respectively, both P>0.05). Conclusions The level of peripheral iNKT cells in HIV infected patients decreases with the disease progression. To certain extent, iNKT cells can reflect the severity of immune damaging in HIV infected patients.

Objective: To investigate the differences of genotyping and virulence of hospital-acquired methicillin-resistant staphylococcus aureus (HA-MRSA) and community-acquired methicillin-resistant staphylococcus aureus (CA-MRSA). Methods: A total of 83 non-repetitive clinical specimens were collected in this study. All samples were classified into HA-MRSA and CA-MRSA according to the definition of the USA Centers for Disease Control and Prevention (CDC). Multiple polyemeras chain reaction (PCR) was used for SCCmec typing, and spa typing was conducted by means of PCR plus DNA sequencing. Panton-valentine leucocidin (PVL), phenol-soluble modulins α (PSMa), phenol-soluble modulins mec (PSM-mec), exfoliative toxin A (ETA), exfoliative toxin B (ETB), toxic shock syndrome toxin (TSST), α-toxin (hla), δ-toxin (hld) were also detected by conventional PCR. Results: The SCCmec type Ⅰ, Ⅱ, Ⅲ, Ⅳa and unclassified type accounted for 1.2%, 3.6%, 65.1%, 28.9% and 1.2%, respectively. 82.1% of the 39 strains of HA-MRSA were identified as SCCmec Ⅲ. While for the 44 strains of CA-MRSA, SCCmec Ⅲ and Ⅳa each accounted for 50%. The proportions of SCCmec Ⅲ and Ⅳa in HA-MRSA and CA-MRSA showed significant differences (χ²=9.343, 20.253, both P<0.05); There were 15 types of spa, t437, t062, t015 accounted for 39.8%, 21.7% and 10.8% respectively. Among the strains of HA-MRSA, the spa type t437 and t062 accounted for 28.2% and 23.1%, respectively, while for the spa type of CA-MRSA, t437 and t062 accounted for 50% and 20.5%, respectively. There was significant difference of t437 type between HA-MRSA and CA-MRSA (χ²=4.100, P=0.043). The positive rates of PSM-a, hla and hld were all 100%, and the positive rates of PVL, ETA, ETB, TSST and PSM-mec were 24.3%, 4.8%, 1.2%, 10.8% and 4.8%, respectively. The proportions of PVL positive HA-MRSA and CA-MRSA were 10.3% and 36.4%, respectively, which was significantly different (χ²=7.705, P=0.006), while PSM-mec positive HA-MRSA and CA-MRSA were 10.3% and 0%, respectively. Conclusions The strains of SCCmec Ⅳa and spa t437 are detected in HA-MRSA, suggesting that the more virulent CA-MRSA genotypes coexist with the traditional HA-MRSA genotypes in the hospital-acquired infections, which brings new challenges for the prevention and control of hospital infection.

Objective: To construct the mutant strain ATP binding cassette transporter SSU05_0948 of Streptococcus suis type 2 and comprehensively study its pathogenicity, and to provide useful insights for understanding the mechanism that Streptococcus suis avoid host innate immunity. Methods: The mutant strain 05ZYH33Δ0948 was constructed through homologous recombination technology. The differences between the mutant strain and the wild type strain were evaluated through bacterial adhesion, whole blood killing, mice meningitis assay, mice and piglets virulence assay. Chi-square test and t test were used. Results: Successfully constructed the mutant strain 05ZYH33Δ0948. The adhesion results showed that the adhesion rate (0.663±0.047)% of the wild strain to A549 cell was significantly higher than that of the mutant (0.246±0.074)%, the difference was statistically significant (χ²=5.267, P=0.014); the adhesion rate (16.540±2.320)% of the wild strain to Hep2 cell was significantly higher than that of the mutant (1.970±0.320)%, the difference was statistically significant (χ²=0.014, P<0.01); the adhesion rate (5.497±0.174)% of the wild strain to Hep2 cell was significantly higher than that of the mutant (1.950±0.335)%, the difference was statistically significant (χ²=0.016, P<0.01). The killing rate (32.970±3.589)% of the wild strain in whole blood is no difference with the mutant (29.560±3.737)% (χ²=1.200, P=0.133). Piglets competitive infection showed that, the competitive index at 12 h, 24 h and 36 h were 0.046±0.003, 0.107±0.003, 0.064±0.001, respectively. 12 h and 24 h was significant differences(t=15.490, P=0.041), 24 h and 36 h was significant differences(t=5.660, P=0.047), 12 h and 36 h was no differences(t=1.445, P=0.285). Conclusions Streptococcus suis type 2 ABC transporter SSU05_0948 is a new adhesion factor and virulence factor of Streptococcus suistype 2, and also a new meningitis factor, which plays important roles in Streptococcus suis against host innate immunity.

Objective: To investigate the etiology composition of enterovirus (EV) in patients with severe hand, foot, and mouth disease (HFMD) in children. To assess the diagnostic value of cerebrospinal fluid (CSF) tests in severe HFMD, and to find the key laboratory tests for severe HFMD. Methods: A total of 288 hospitalized cases of children clinically diagnosed with severe HFMD in Hangzhou Children′s Hospital were included from March to July 2016. Throat swabs were collected and enterovirus nucleic acids were detected by fluorescence quantitative reverse transcription (RT)-PCR. Synchronous CSF and serum samples were collected for EV-A71 and CV-sackievirus A16 (CV-A16)-IgM antibody detection. CSF samples underwent routine and biochemical tests. Normally distributed continuous variables were compared using t test. Non-normal distribution continuous variables were compared using Mann-Whitney U test. Differences between categorical variables were compared with χ² test. Results: The total positive rate of enterovirus nucleic acid EV-A71/CV-A16/EV by fluorescence quantitative RT-PCR in the 288 cases of children clinically diagnosed with HFMD was 83.7% (241/288), including EV-A71 55.2% (159/288), CV-A16 4.9% (14/288) and the other enterovirus 23.6% (68/288). Among the other enterovirus group, there were 29.4% CV-A6 (20/68) , 16.2% CV-A4 (11/68) and CV-A10 10.3% (7/68). The total positive rate of combined serum and CSF detection of EV nucleic acid and EV-A71 and CV-A16 IgM antibody was 98.3% (283/288). EV nucleic acid positive rate was 83.7% (24/288). The positive rates were statistically different (χ² =37.289, P=0.000). The CSF nucleated cells count in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-4.472 and -9.991, respectively, both P<0.05). The CSF nucleated cells positive rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=43.857 and 133.078, respectively, both P<0.05). The CSF protein level in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (Z=-3.151 and -5.255, respectively, both P<0.05). The CSF protein positive (>400 mg/L) rate in EV-A71 positive subgroup was higher than those in CV-A16 positive subgroup and other enteroviruses subgroup (χ²=4.956 and 11.795, respectively, both P<0.05). The CSF nucleated cell counts and positive rates in EV-A71 IgM antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The CSF protein level and elevated proportion in antibody-positive subgroup were both higher than those in antibody-negative subgroup (both P<0.05). The lactate dehydrogenase concentration in antibody-positive subgroup was significantly higher than those in antibody-negative subgroup (P<0.05). The EV-A71 IgM antibody in serum was significantly correlated with the antibody in CSF (r=0.600, P<0.05). Conclusions EV-A71 is still the most important pathogen of severe HFMD in Hangzhou in 2016. Other enterovirus such as CV-A6, CV-A4 increases compared to those in 2014 and 2015. The CSF routine and biochemical tests and the IgM antibody levels can serve as an important indicator for the diagnosis of children with severe HFMD.