Evaluation of the exposure risk of SARS-CoV-2 in different hospital environment.

The ongoing coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has imposed a significant impact on social and economic activities. As a high infectious pathogen, the existence of SARS-CoV-2 in public space is very important for its transmission. During the COVID-19 pandemic, hospitals are the main places to deal with the diseases. In this work, we evaluated the exposure risk of SARS-CoV-2 in hospital environment in order to protect healthcare workers (HCWs). Briefly, air and surface samples from 6 different sites of 3 hospitals with different protection levels were collected and tested for the SARS-CoV-2 nucleic acid by reverse transcription real-time fluorescence PCR method during the COVID-19 epidemic. We found that the positive rate of SARS-CoV-2 nucleic acid was 7.7 % in a COVID-19 respiratory investigation wards and 82.6 % in a ICUs with confirmed COVID-19 patients. These results indicated that in some wards of the hospital, such as ICUs occupied by COVID-19 patients, the nucleic acid of SARS-CoV-2 existed in the air and surface, which indicates the potential occupational exposure risk of HCWs. This study has clarified retention of SARS-CoV-2 in different sites of hospital, suggesting that it is necessary to monitor and disinfect the SARS-CoV-2 in hospital environment during COVID-19 pandemic, and will help to prevent the iatrogenic infection and nosocomial transmission of SARS-CoV-2 and to better protect the HCWs.

PBEF promotes the apoptosis of pulmonary microvascular endothelial cells and regulates the expression of inflammatory factors and AQP1 through the MAPK pathways.

Pre-B cell colony-enhancing factor (PBEF) has been shown to have a variety of biological functions. Studies have proven that PBEF plays a functional role in acute lung injury (ALI). Therefore, in this study, we aimed to confirm the importance of PBEF in ALI. The effects of PBEF overexpression on the apoptosis of human pulmonary microvascular endothelial cells (HPMECs) were analyzed by flow cytometry, and the results indicated that PBEF promoted the apoptosis of HPMECs, which aggravated the development of ALI. Comparative experiments involving increasing and decreasing PBEF expression demonstrated that PBEF promoted the expression of inflammatory factors, such as interleukin (IL)­1ß, IL­6 and IL­8 in the HPMECs , thus intensifying the inflammatory response. PBEF also inhibited the expression of aquaporin 1 (AQP1), which caused a dysfunction and imbalance in water transport. Moreover, we also found that tumor necrosis factor (TNF)­α promoted the expression of PBEF in the HPMECs. After blocking the mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) pathways, we found that PBEF regulated the expression of inflammatory factors and AQP1, mainly through the MAPK pathways. Taken together, these results demonstrate that the increase in intracellular PBEF expression promoted the apoptosis of HPMECs and the expression of inflammatory factors and thus enhanced the inflammatory response and inhibited the expression of AQP1, which resulted in abnormal water transport, diminishing the regulatory effects of AQP1 on water transport.

JAZF1 regulates visfatin expression in adipocytes via PPARα and PPARß/δ signaling.

OBJECTIVE: Current whole genome-wide association study has identified the association of JAZF1 with type 2 diabetes; its close relation with glucose and lipid metabolism has also been revealed. However, to date, JAZF1 remains a relatively new gene with unknown function. MATERIALS/METHODS: We constructed JAZF1 overexpression vector and synthesized JAZF1 siRNA, then transfected them into 3T3-L1 adipocytes, investigated the relationship between the regulations of JAZF1, visfatin, and other adipokines, researched the specific function of JAZF1 in glucose and lipid metabolism. RESULTS: This study found that the expression of JAZF1 was gradually but significantly upregulated during the induced differentiation of 3T3-L1 preadipocytes, and that the trend of its expression was consistent with that of visfatin. Further studies indicated that JAZF1 promoted the expressions of visfatin, PPARα, and PPARß/δ in adipocytes but simultaneously inhibited the expressions of TAK1 and PPARγ. Luciferase reporter assay revealed that JAZF1 activated the transcription of visfatin, but ChIP assay results indicated that JAZF1 did not directly bind to visfatin PPRE. Our results also showed that the JAZF1 overexpression-increased visfatin expression was abolished by the addition of PPARα antagonist GW 6471 and PPARß/δ antagonist GSK 3787 respectively. And these results were further confirmed by the experiment with PPARα and PPARß/δ siRNAs. Meanwhile, we also found that JAZF1 inhibited the lipid accumulation during the differentiation of 3T3-L1 into mature adipocyte. CONCLUSIONS: Our results indicate that JAZF1 might firstly upregulated the expression of PPARα and PPARß/δ, which in turn activated the transcription of visfatin. JAZF1 plays an important role in lipid metabolism and may thus provide a potential tool for the treatment of obesity and lipid metabolism disorders among other diseases.

[Effects of simvastatin on lipopolysaccharide induced α-subunit epithelial sodium channel mRNA in rat lung alveolar type II epithelial cells].

OBJECTIVE: To study the impact of simvastatin on α-subunit epithelial sodium channel (α-ENaC) mRNA expression in primary culture alveolar typeII (ATII) epithelial cell of rats induced by lipopolysaccharide (LPS) in vitro. METHODS: ATII of primary generation were isolated from adult Sprague-Dawley (SD) rats. The cells were randomly divided into five groups: blank control group, LPS injured group (final concentration of LPS 1 mg/L), simvastatin low and high concentration groups (final concentration of simvastatin 20 µmol/L, 30 µmol/L, respectively), solution control group. Then, after being intervened for 1, 12 and 24 hours, the level of human tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) were monitored by enzyme-linked immunosorbent assay (ELISA), and α-ENaC mRNA expression was tested by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: After being intervened for 1, 12 and 24 hours, expressions of TNF-α and IL-1ß in LPS injured group were obviously higher than those in blank control group. Expressions of TNF-α and IL-1ß at 1, 12 and 24 hours in simvastatin low concentration group were significantly decreased compared with those in LPS injured group (TNF-α 1 hour: 1178.80±127.43 ng/L vs. 2336.00±170.04 ng/L, 12 hours: 1003.60±59.61 ng/L vs. 2479.80±210.41 ng/L, 24 hours: 695.80±25.24 ng/L vs. 1167.60±132.72 ng/L; IL-ß 1 hour: 285.00±42.60 ng/L vs. 429.60±27.39 ng/L, 12 hours: 238.60±24.12 ng/L vs. 822.20±12.74 ng/L, 24 hours: 213.40±17.87 ng/L vs. 637.60±22.96 ng/L, all P<0.05). Expressions of TNF-α and IL-1ß in high concentration group were decreased more obviously than those in low concentration group (TNF-α 1 hour: 965.60±24.45 ng/L vs. 1178.80±127.43 ng/L, 12 hours: 522.80±16.89 ng/L vs. 1003.60±59.61 ng/L, 24 hours: 252.40±17.64 ng/L vs. 695.80±25.24 ng/L; IL-1ß 1 hour: 225.60±34.44 ng/L vs. 285.00±42.60 ng/L, 12 hours: 190.60±17.64 ng/L vs. 238.60±24.12 ng/L, 24 hours: 152.80±14.70 ng/L vs. 213.40±17.87 ng/L, all P<0.05), but increased compared with those in blank control group. After being intervened for 1 hour, no evident changes were observed in expression of α-ENaC mRNA in all groups. After being intervened for 12 hours and 24 hours, evident decrease in expression of α-ENaC mRNA (A value) was observed in LPS injured group compared with blank control group (12 hours: 0.211±0.021 vs. 0.496±0.027, 24 hours: 0.253±0.030 vs. 0.482±0.030, both P<0.05). Expressions of α-ENaC mRNA in simvastatin low concentration group evidently increased compared with those in LPS injured group (12 hours: 0.363±0.030 vs. 0.211±0.021, 24 hours: 0.309±0.024 vs. 0.253±0.030, both P<0.05). Expressions of α-ENaC mRNA in simvastatin high concentration group increased more obviously compared with those in low concentration group (12 hours: 0.413±0.034 vs. 0.363±0.030, 24 hours: 0.346±0.024 vs. 0.309±0.024, both P<0.05), but decreased compared with blank control group. No evident difference in expressions of all indexes in solution control group was observed compared with those in blank control group. CONCLUSIONS: High dose simvastatin could improve α-ENaC mRNA expression in primary culture ATII epithelial cells of rats. This may act by modulation the level of TNF-α and IL-1ß.

Prognostic indicators of patients with acute kidney injury in intensive care unit.

BACKGROUND: Acute kidney injury (AKI) is associated with a high mortality. This study was undertaken to detect the factors associated with the prognosis of AKI. METHODS: We retrospectively reviewed 98 patients with AKI treated from March 2008 to August 2009 at this hospital. In these patients, 60 were male and 38 female. Their age ranged from 19 to 89 years (mean 52.4±16.1 years). The excluded patients were those who died within 24 hours after admission to ICU or those who had a history of chronic kidney disease or incomplete data. After 60 days of treatment, the patients were divided into a survival group and a death group. Clinical data including gender, age, history of chronic diseases, the worst laboratory values within 24 hours after diagnosis (values of routine blood tests, blood gas analysis, liver and renal function, levels of serum cystatin C, and blood electrolytes) were analyzed. Acute physiology, chronic health evaluation (APACHE) II scores and 60-day mortality were calculated. Univariate analysis was performed to find variables relevant to prognosis, odds ratio (OR) and 95% confidence interval (CI). Multiple-factor analysis with logistic regression analysis was made to analyze the correlation between risk factors and mortality. RESULTS: The 60-day mortality was 34.7% (34/98). The APACHE II score of the death group was higher than that of the survival group (17.4±4.3 vs. 14.2±4.8, P<0.05). The mortality of the patients with a high level of cystatin C>1.3 mg/L was higher than that of the patients with a low level of cystatin C (<1.3 mg/L) (50% vs. 20%, P<0.05). The univariate analysis indicated that organ failures≥2, oliguria, APACHE II>15 scores, cystatin C>1.3 mg/L, cystatin C>1.3 mg/L+APACHE II>15 scores were the risk factors of AKI. Logistic regression analysis, however, showed that organ failures≥2, oliguria, cystatin C>1.3 mg/L +APACHE II>15 scores were the independent risk factors of AKI. CONCLUSION: Cystatin C>1.3 mg/L+APACHE II>15 scores is useful in predicting adverse clinical outcomes in patients with AKI.

[Effect of intrathecal pumping tramadol on the immune function in rats with formalin pain].

OBJECTIVE: To evaluate the effect of intrathecal pumping tramadol on cell-mediated immunity in rats with formalin inflammatory pain. METHODS: Thirty-two Sprague-Dawley adult male rats weighting 250 approximately 300 g were randomly divided into 4 groups (n=8 in each group):Saline group (NS) and 3 tramadol groups (T1,T2,and T3). The rats were anesthetized with intraperitoneal chloral hydrate (300 approximately 350)mg/kg. Microspinal catheter was inserted into the subarachnoid space at the lumber region according to modified Yaksh techniques. In the tramadol groups,after 5 days tramadol was continuously infused through the spinal catheter at 50 (T1),25 (T2), and 12.5 microg/h (T3) for 7 days. In the NS group normal saline was continuously infused instead of tramadol. On Day 7 formalin (5%, 50 microL) was injected into the plantar surface of the left hindpaw. The number of flinches, lickings and total time of licking was recorded for 60 min.Pain intensity scoring(PIS)(0 approximately 3;0= no pain, 3=severe pain) was used to assess the antinociceptive effect of intrathecal tramadol. The rats were killed after the evaluation of pain intensity. Body weight and spleen weight were measured and spleen index (spleen weight/body weight) was calculated. T-lymphocyte function was evaluated based on Concanavalin-A(ConA) induced splenocyte proliferation. A modified lactic acid dehydrogenase(LDH) release assay was done to assess the NK cell activity. Phenotypic expressions of cell surface markers of T lymphocyte subsets (CD3+, CD3+ CD4+, CD3+ CD8+, and CD4+/ CD8+) and NK cell(CD161+) in the spleen were analyzed by flow cytometry. RESULTS: The PIS scores were significantly lower in the T1,T2,and T3 groups than those in the NS group. The spleen index and splenocyte proliferation induced by ConA were significantly suppressed in the T1 group,and the phenotypes of T lymphocyte subsets were significantly changed,but no significant difference was found in the T2 and T3 groups compared with the NS group. There were no differences in NK cell activity in the 3 tramadol groups from the control group. CONCLUSION: Intrathecal pumping tramadol has significantly antinociceptive effect. Intrathecal pumping higher dosage tramadol (50microg/h) suppresses T lymphocyte proliferation and alteres T lymphocyte subset phenotype but does not affect NK cell activity. General analgesic dosage tramadol (25 and 12.5 microg/h) has no effect on the immune function.

[Protective effect of lidocaine on injury alveolar Type II cells induced by LPS in adult rats].

OBJECTIVE: To investigate the effect of lidocaine on LPS induced apoptosis of cultured adult rat alveolar Type II (AT-II) cells. METHODS: Cultured cells were exposed to LPS and lidocaine for 24 hours. Apoptosis and necrosis rates of cells were detected by flow cytometry and electron microscope. The activity of lactic dehydrogenase (LDH) was analyzed by using LDH kits. RESULTS: LPS induced the AT-II cell injuries by increasing not only the necrosis and apoptosis rates but also the LDH release of cultured AT-II in vitro. Lidocaine decreased the necrosis and apoptosis rates of AT-II cells. CONCLUSION: Lidocaine can directly inhibit the apoptosis and necrosis induced by LPS in cultured AT-II cells.

[Clinical study of low molecular weight heparin therapy for sepsis].

OBJECTIVE: To investigate the therapeutic effect of low molecular weight heparin (LMWH) therapy on sepsis. METHODS: Forty sepsis patients were randomly divided into two groups: routine treatment group and LMWH treatment group. Score of acute physiology and chronic health evaluation II (APACHE II), the days in intensive care unit (ICU) and mortality rate in 28 days were observed, and the levels of interleukin-6 (IL-6), malondialdehyde (MDA), superoxide dismutase (SOD), coagulation function and platelet count (PLT) were determined before and after treatment in the two groups. RESULTS: Both APACHE II and IL-6 levels in LMWH group decreased with passage of time, the differences were significant between the results on day 7 and that of pretreatment (both P<0.05). In the routine treatment group, APACHE II and IL-6 levels decreased first and then increased, and they were higher than those in LMWH group 7 days after treatment (both P<0.05). In LMWH group, the time of stay in ICU was (9.92+/-6.81)days, the mortality rate in 28 days was 40.9%, and they all were lower than those in routine treatment group [(12.85+/-9.14)days and 50.0%], but the difference was not statistically significant (both P>0.05). After treatment SOD level elevated [(159.13+/-99.31) kU/L vs.(318.38+/-284.29) kU/L] and MDA level lowered [(17.72+/-14.89) micromol/L vs.(6.62+/-5.53) micromol/L] in LMWH group. The changes in MDA and SOD in routine treatment were reverse to those of the LMWH group [SOD: (180.99+/-169.40) kU/L vs. (135.16+/-107.73) kU/L; MDA: (17.25+/-15.74) micromol/L vs. (20.77+/-16.87) micromol/L]. The difference was significant between the two groups after treatment (both P<0.05). The difference in coagulation function and PLT was not significant between the two groups. CONCLUSION: LMWH can ameliorate sepsis by down-regulating the levels of pro-inflammatory cytokines, and suppressing the release of oxygen-derived free radicals. It is a promising treatment measure in sepsis patient with safety and no severe side effects.