Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

In the case of intracranial hemorrhage,coders tend to ignore the cause of intracranial hemorrhage in the cod-ing,whether it is spontaneous intracranial hemorrhage or intracranial hemorrhage caused by trauma,and the coding of the two is completely different in ICD-10.The former is classified as I60-I62 while the latter is classified as S06.Different etiology will also enter different DRG groups when DRG is included.When determining the cause,the site of intracranial hemorrhage should be determined whether it is subarachnoid hemorrhage,or epidural/subdural hemorrhage or cerebral parenchymal hemorrhage,be-cause different bleeding sites have different codes in ICD-9-CM-3 when performing blood removal in cranial swelling.The classifi-cation of epidural hematoma removal was on 01.24,subdural or subarachnoid hematoma removal was on 01.31,and intracerebral parenchymal hematoma removal was on 01.39.The removal of intracranial hematoma is usually divided into cone craniotomy,skull trepanation and drainage and traditional craniotomy according to different operation methods.The operation process of these three operations is obviously different,and coders need to understand the characteristics of the three operations to achieve accurate classification.In the DRG grouping,the disease code is different from the surgical code and the DRG group will be different.Through understanding the definition and etiology of intracranial hematoma removal,the coding ideas of intracranial hematoma re-moval were analyzed,so as to improve the professional ability of coders and ensure the accuracy of DRG data.

Objective To predict the prognosis of patients after cardiac surgery by implementing hemodynamic monitoring collective strategy—CHOLKIT protocol,and to explore the practical application value of CHOLKIT protocol,such as early intervention to improve tissue circulation and microcirculation,and predict the timing of extubation.Methods A prospective cohort study was used to analyse 88 patients who underwent cardiac surgery in the First Affiliated Hospital of Xinjiang Medical University from April to October 2020,and they were divided into the survival group (84 cases) and the death group (4 cases) with patient survival or death as the study endpoint.The CHOLKIT protocol was applied to score the central venous pressure (CVP),heart rate (HR),central venous oxygen saturation (ScvO2),lactic acid (Lac),potassium (K+),perfusion index (PI),and toe temperature (T) of patients at different time periods.The correlation between CHOLKIT score and prognosis was predicted based on the scores.Results Some monitoring indexes in the CHOLKIT protocol were related to the mortality,renal injury and duration of mechanical ventilation after cardiac surgery.Conclusion The CHOLKIT protocol can predict the timing of extubation and the change of the condition of patients after cardiac surgery,and early intervention can improve the prognosis of patients,duration of mechanical ventilation and number of days of stay in the intensive care unit.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*

Objective:To evaluate whether underdilated stent could reduce the occurrence of hepatic encephalopathy (HE) after transjugular intrahepatic portosystemic shunt (TIPS) creation.Methods:A total of 197 patients with decompensated liver cirrhosis, who had underwent TIPS creation at Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, were analyzed retrospectively, including 110 males and 87 females with age 25-79 (54±11) years old. Uncovered and covered stents with 8 mm diameter were implanted in all subjects, and then dilated by balloon catheters with 6 mm or 8 mm diameter. The patients were divided into two groups, including underdilated group (6 mm, n=105) and control group (8 mm, n=92).Kaplan-Meier curves were used to illustrate cumulative rate of HE, and the differences were assessed with the log-rank test. Multivariate analyses with a Cox regression model were conducted to explore the risk factors for HE. Results:During a median follow-up period of 29 (12-54) months, 16 (15.2%) patients developed HE in the underdilated group and 27 (29.3%) patients in the control group. There was a significant difference in the cumulative rate of HE ( P=0.014), but no statistical differences were found in terms of variceal rebleeding, shunt dysfunction and survival between the two groups ( P=0.608, P=0.659, P=0.968). In multivariated analysis, group assignment (underdilated vs. control, HR=0.291, 95% CI 0.125-0.674, P=0.004) was identified as an independent risk factor for HE after TIPS creation. Conclusion:Underdilated TIPS could reduced the risk of HE compared with completely dilated TIPS, with comparable risk of variceal rebleeding, shunt dysfunction and mortality. And it is worthy of applying this technique to a large sample of patients in clinical practice.

Objective:To explore the effect of enriched environment on pain sensitivity, anxiety- and depressive-like behavior in selective nerve injury(SNI) rats model and its potential mechanism.Methods:A total of 36 male clean grade SD rats aged 6-8 weeks were randomly divided into three groups( n=12 in each group): sham operation+ standard environment group (sham group), SNI+ standard environment group (standard environment group), SNI+ enriched environment group (enriched environment group). The rat model of neuropathic pain was established by SNI.The rats in the enriched enviroment group were placed in an enriched enviroment 7 days before operation until 21 days after operation.The paw withdraw threshold(PWT) and paw withdraw latency (PWL) were performed to assess hyperalgesia.The open field test, elevated plus maze test, novelty suppressed feeding test and forced swimming test were used to assess anxiety and depression like behavior.The expressions of cAMP response element binding protein (CREB), p-CREB, brain-derived neurotrophic factor (BDNF), postsynaptic density-95 (PSD-95) and neuroligin 2 (NLGN2) were detected by Western blot.The expression of CREB and BDNF in contralateral ACC were measured by immunofluorescence.GraphPad prism 8.0 and SPSS 23.0 were used for data analysis.One way ANOVA was used for inter group comparison, repeated measurement ANOVA was used to analyze PWT and PWL results, and Tukey test was used for pairwise comparison. Results:(1) In PWT and PWL experiments, the interaction effect between group and time, group main effect and time main effect of PWT were significant ( F=13.4, 39.6, 369.6, all P<0.05), and the interaction effect between group and time, group main effect and time main effect of PWL were significant ( F=3.8, 10.3, 58.8, all P<0.05). Compared with sham group, PWT((8.0±3.5) g, (2.4±1.4) g, (2.3±1.1) g, (2.2±1.6) g, (1.6±0.5) g) and PWL((8.6±1.3) s, (7.3±1.5) s, (7.9±1.0) s, (6.6±1.1) s, (7.7±1.4) s) in standard environment group decreased at each time point (all P<0.05). (2) Compared with sham group, the number of entrying into the central area (1.3±1.7), the time of entrying into the central area((1.6±1.3) s), the proportion of entering open arms ((8.0±7.8) %) and the proportion of time in the open arms ((1.3±1.2) %) all significantly decreased in standard environment group ( t=4.585, 5.423, 4.682, 5.202, all P<0.05). The eating latency ((365.2±94.4) s) and immobility time ((127.6±24.3) s) dramatically increased ( t=6.008, 14.290, both P<0.05). The number and time of entrying into central area of enriched environment group were both higher than those of standard environment group(both P<0.05), while the eating latency and immobility time of enriched environment group were both lower than those of standard environment group(both P<0.05). (3) Compared with sham group(CREB: (1.6±0.2), (0.8±0.5); BDNF: (0.8±0.5), (1.0±0.4)), the expression of CREB ((1.8±0.1), (1.5±0.2)), BDNF ((0.9±0.6), (1.4±0.3)) in spinal cord and ACC of standard environment group increased (spinal: t=3.283, 4.989; ACC: t=5.502, 4.257, all P<0.05). The expression of PSD-95 ((1.6±0.2), (1.0±0.2) and NLGN2 ((1.5±0.5), (1.1±0.2)) also increased in ACC of standard enviroment group ( t=4.257, 2.214, both P<0.05). Compared with standard environment group, the expression of CREB (1.3±0.3), BDNF (0.7±0.4), PSD-95(1.0±0.3) and NLGN2(1.1±0.4) in spinal cord of enriched environment group decreased ( t=5.007, 2.166, 2.358, 2.322, all P<0.05). The expression of PSD-95(1.2±0.3) and NLGN2(1.1±0.2) also decreased in ACC of enriched environment group ( t=2.674, 2.944, both P<0.05). However, the expression of p-CREB (1.7±0.6) and BDNF (2.4±0.2) increased in ACC ( t=4.180, 7.610, P<0.05). Conclusion:Enriched environment can improve neuropathic pain and anxiety- and depressive-like behavior in SNI rats, which may be related to the change of synaptic plasticity in spinal cord and ACC.

Microcystis aeruginosa, a type of algal bloom microalgae, is widely distributed in water, causing serious deteriorated effects on humans and the ecological environment. As a biocontrol microorganism, Bacillus subtilis can synthesize various bioactive substances through non-ribosomal peptide synthetase, to inhibit the growth of M. aeruginosa. Thus, it is imperative to investigate the non-ribosomal peptide (NRP) metabolites of B. subtilis fmb60. Three NRP metabolites from B. subtilis fmb60 including bacillibactin, surfactin and fengycin were extracted and identified by genome mining technology. The growth inhibition of M. aeruginosa was studied by adding various concentrations of NRP metabolites. The half-effect concentration value (EC50.4 d) of M. aeruginosa was 26.5 mg/L after incubation for 4 days. With the increasing concentration, the inhibitory effects of NRP metabolites of B. subtilis fmb60 on M. aeruginosa was enhanced significantly. Compared with the control group, with the addition of 50 mg/L NRP metabolites to the M. aeruginosa, the content of Fv/Fm, Fv/Fo and Yield parameter after cultured for 4 days were decreased by 2.8%, 1.7% and 2.0%, respectively. Those findings indicate that the NRP metabolites of B. subtilis fmb60 can significantly inhibit the photosynthesis and metabolism of M. aeruginosa, which provides a theoretical foundation for the development of biological algae inhibitor of B. subtilis.
Bacillus subtilis ; Humans ; Microcystis ; Peptides ; Photosynthesis

Objective:To determine the content of albiflorin, paeoniflorin, benzoylpaeoniflorin, sophoricoside, prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside and sec-O-glucosylhamaudol in Huaiqin Salve with high performance liquid chromatography-quantitative analysis of multi-components by single-marker (HPLC-QAMS) method. Methods:The samples were separated with ZORBAX Eclipse XDB-C18 column by a gradient elution using methanol-acetonitrile (1:2) (A)-0.1% phosphoric acid solution (B) as mobile phase and the flow rate was 1.0 ml/min. The detection wavelength was 230 nm for albiflorin, paeoniflorin and benzoylpaeoniflorin, and 254 nm for sophoricoside, prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside and sec-O-glucosylhamaudol. The column temperature was 30 ℃. Using sophoricoside as an internal standard, the relative correction factors (RCFs) of albiflorin, paeoniflorin, benzoylpaeoniflorin, prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside and sec-O-glucosylhamaudol were calculated. The method was validated by comparison of the quantitative results between external standard method (ESM) and HPLC-QAMS method.Results:Albiflorin, paeoniflorin, benzoylpaeoniflorin, sophoricoside, prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside and sec-O-glucosylhamaudol showed good linear relationship within the range of 0.034 7-0.867 5, 0.063 6-1.590 0, 0.006 9-0.172 5, 0.198 6-4.965 0, 0.092 8-2.320 0, 0.026 6-0.665 0, 0.042 7-1.067 5, 0.020 9-0.522 5 μg ( r ≥ 0.999 1); whose average recoveries ( RSDs) were 98.85% (1.02%), 100.04% (0.67%), 96.92% (1.14%), 100.06% (0.85%), 99.31% (1.39%), 99.16% (1.22%), 98.59% (1.33%) and 97.58% (1.41%), respectively. No significant differences were found in the quantitative analysis of the components with ESM and HPLC-QAMS method. Conclusion:The HPLC-QAMS method could provide the reference for themulti-index component evaluation for Huaiqin Salve.

Objective:To assess the association between the different prehospital transfer pathways to the hospital and reperfusion delay in patients with ST segment elevation myocardial infarction (STEMI).Methods:We retrospective collected 320 STEMI patients aged 18 years or older who underwent primary percutaneous coronary intervention (PPCI) from June 1, 2016 to July 31, 2018. They were divided into three groups according to different prehospital transfer pathways: patients directly transferred from the field by ambulance to PCI-capable center hospital (field transfer group, n=29); patients transferred by ambulance from PCI-incapable hospitals to PCI-capable center hospital (inerhospital transfer group, n=111); patients transferred by friends or relatives to PCI-capable center hospital (self-transfer group, n=180). The basic characteristic attributes, reperfusion time and fatal complications such as acute left heart failure cases (ALHF) cases, ventricular fibrillation (VF) cases, and in hospital death were collected and compared. In addition, logistic regression analysis was used to analyze uni- and multivariate of door-to-balloon (D2B) time less than 90 min. Results:The S2FMC interval were 118 min (50, 377) min , FMC-to-balloon interval were 87 min (66, 120.5) min and the onset-to-balloon time were 221 min (135, 482.5) min. The above three interval in the interhospital transfer group were longer than those in the field transfer and self-transfer groups ( P<0.05). S2FMC accounted for 51.14% of onset-to-balloon time in the field transfer group, 63.29% in the interhospital transfer group and 55.26% in the self transfer group. The door-to-catheter room interval were 33 min (9, 53.5) min. The interval in the interhospital transfer group were shorter than those in the field transfer and self-transfer groups ( P<0.05). The interval in the self-transfer group were longer than those in the interhospital transfer and field transfer groups ( P<0.05). Multiple regression analysis showed that the interhospital transfer group ( OR=15.251, 95% CI: 5.328-43.657, P<0.01), field transfer group ( OR=8.219, 95% CI: 1.861-36.307, P=0.005), FMC2ECG time ( OR=0.975, 95% CI: 0.962-0.989, P<0.01), and smoking ( OR=2.099, 95% CI: 1.015-4.341, P=0.045) were independent predictor of goal time less than 90 min ( P<0.05 for all variables). Adverse events in STEMI patients mainly occurred within 6 h of the initial symptoms. The incidence of VF was 3.75% (95% CI: 3.73%-3.77%), ALHF was 10.94% (95% CI: 7.52%-14.36%) and in-hospital mortality was 2.5% (95% CI: 0.79%-4.21%). There was no significant difference in the incidence of adverse events among the three groups ( P>0.05). Conclusions:The symptoms of STEMI patients are at risk in the early stage. Ambulances can shorten the reperfusion time, but the ambulance system is less used by patients. It is necessary to further optimize the treatment process of non-PCI hospitals and strengthen the popularity of emergency knowledge related to chest pain among residents, so as to shorten the reperfusion time.