Objective To establish a HPLC method for the determination of gallic acid in Radix et Rhizoma Rhei and to study the changes of gallic acid content in Radix et Rhizoma Rhei during processing. Methods HPLC method was used to detect gallic acid content. Diamonsil C18(250 mm?4.6 mm,5?m) column was used,and the mobile phase was a mixed liquid of MeOH -0.01 %H3PO4(10∶90). The column temperature was set up at 30℃,the flow rate was 1 mL?min-1,and the detecting wave-length was 273 nm. Results There were obvious differences of gallic acid content between the crude herbal material and different kinds of processed products of Radix et Rhizoma Rhei. The content of gallic acid was decreased in Radix et Rhizoma Rhei prepared by wine,but was increased in Radix et Rhizoma Rhei prepared by steaming with wine and by stewing with wine,and in charred Radix et Rhizoma Rhei. Conclusion The different processing methods have certain effect on the content of gallic acid in Radix et Rhizoma Rhei.

ObjectiveTo investigate the feasibility and safety of Proseal-laryngeal mask airway(PLMA) for anesthesia in endoscopic thyroidectomy.MethodsForty ASA class Ⅰ or Ⅱ patients underwent endoscopic thyroidectomy via breast approach were randomly divided into laryngeal mask venting group( Group P,n =20) and endotracheal tube venting group( Group T,n =20).Both groups were used the same anesthesia induction,maintenance and breathing parameter adjustment.The HR and MAP in both groups were recorded respectively before intubation,while intubation,2 min after intubation,before and after head back out center position,before extubation and 2 min after exbutation.The adverse effects of respiratory tract were recorded respectively.ResultsThere were no significant changes in HR and MAP in Group P before and after intubation,as well as before and after extubation( all P ＞ 0.05 ) ;In Group T,the HR and MAP while intubation,2 min after intubation are higher than that before intubation (all P ＞ 0.05).The HR and MAP in Group T,2 min after exbutation are higher than before extubation(all P ＞ 0.05 ).The incidence rate of sore throat and hoarse postoperatively were lower in Group P than in Group T( all P ＞ 0.05 ).ConclusionPLMA could be safely and effectively used in the anesthesia of endoscopic thyroidectomy.

Objective To investigate the trend of liver function changes in untreated adult patients with Graves’ disease in China, and to analyze the associated factors. Methods Patients with newly diagnosed as well as recurrent Graves’ disease from January 2006 to August 2014 were enrolled. They were over 18 years old and did not receive any treatments, Examination of liver function, thyroid function, and thyroid related antibodies as well as tests regarding virus hepatitis were performed. Results A total of 1 254 patients were enrolled. 33 patients with virus hepatitis were ruled out. Ultimately, 1 221 patients matched the criteria of our trial, with 347 males and 874 females [(39. 3 ± 9. 5) year old]. After inclusion, they were assigned to 2 groups according to their liver function results(605 in normal group and 616 in abnormal group). Compared to normal group, patients in the abnormal group were older [(40. 1 ± 9. 2 vs 38. 5 ± 8. 7) year old, P<0. 05] and with higher proportion of females(81. 8% vs 61. 2% , P<0. 05). Regarding the thyroid function and related antibody tests, some patients yielded results that were extremely high so as to exceed the upper limit of the normal range. These patients were more frequently seen in the group with abnormal liver function. The patients whose thyroid function parameters exceeded the upper limit had higher level of alanine aminotransferase[ALT,(37. 69 ± 7. 51 vs 31. 90 ± 5. 95) U/ L, P<0. 05], aspartate aminotransferase[AST, (31. 97 ± 5. 09 vs 27. 88 ± 3. 82) U/ L, P<0. 05], direct bilirubin[DBiL, (5. 58 ± 0. 77 vs 4. 54 ± 0. 71) μmol/ L, P<0. 05]than the group whose thyroid function on the detected range. In the patients with all results detected, patients in abnormal liver function group had higher level of triiodthyronine[T3 , (5. 42 ± 0. 29 vs 4. 94 ± 0. 33) nmol/ L, P<0. 05], thyroxin[T4 ,(217. 53 ±14. 32 vs 204. 22 ±13. 54) nmol/ L, P<0. 05], free triiodthyronine[FT3 ,(15. 88 ± 2. 86 vs 14. 48 ±4. 83) pmol/ L, P<0. 05], free thyroxin[FT4 ,(48. 91 ±8. 45 vs 42. 95 ±6. 14) pmol/ L, P<0. 05], thyroid peroxidase antibody[ TPOAb, (402. 75 ± 89. 99 vs 210. 70 ± 44. 63) IU/ ml, P < 0. 05] and thyrotrophin receptor antibody[TRAb,(14. 08 ± 5. 24 vs 9. 04 ± 2. 58) IU/ L, P<0. 05]. Multivariate logistic regression analysis revealed that patients’ age(OR=0. 98, 95% CI 0. 97-0. 99), gender(OR=0. 94, 95% CI 0. 91-0. 97), level of FT4 (OR=3. 08, 95% CI 2. 19-4. 32), TPOAb(OR = 0. 98, 95% CI 0. 97-0. 99), and TRAb(OR = 1. 07, 95% CI 1. 01-1. 12) were independent risk factors of their liver dysfunction. Conclusion Graves’ disease may lead to liver dysfunction, which is much more common and severe in elder and female patients, as well as patients who are suffering from hyperthyroidism and raised level of thyroid related antibodies.

Objective To investigate the correlation of plasma interleukin ( IL)-17 level and IL-17 receptor (IL-17R) expression in the thymus of patients with myasthenia gravis (MG).Methods The blood samples of 63 patients (38 with glucocorticoid treatment, 25 with thymus removal) who admitted to Henan Provincial People′s Hospital between 2010 and 2014 were collected at three different stages: pre-treatment, 1 week post-treatment and 1 month post-treatment.The blood samples of 42 healthy controls were also collected.Enzyme linked immunosorbent assay was used to evaluate the levels of IL-17 in plasma.Twenty-five thymus tissues from MG patients and another 12 thymus tissues from patients with congenital heart disease who had surgery therapy were also collected.Reverse transcription polymerase chain reaction was used to evaluate the mRNA levels of IL-17R.The possible correlation between the expression of IL-17 and IL-17R with MG was analyzed.Results Before treatment, the levels of IL-17 in the plasma were much higher in all the MG patients ( both ocular and generalized) when compared to the healthy controls ( controls (3.2 ±0.7) pg/ml, MG patients (8.5 ±1.7) pg/ml, t =2.450, P <0.01; generalized type patients (9.7 ±1.4) pg/ml, t =2.532, P <0.01).In the patients with glucocorticoid treatment, IL-17 levels began to reduce after 1 week treatment and a statistically significant difference was found when compared to the pre-treatment samples (pre-treatment (8.3 ±1.2) pg/ml, 1 week after treatment (6.3 ±0.7) pg/ml, t=2.052, P<0.05) and healthy controls (t =1.933, P<0.05).One month after the glucocorticoid treatment, the levels of IL-17 decreased to the normal level (1 month after treatment (3.9 ±0.6) pg/ml, t=2.630, P <0.01, compared to the pre-treatment; t =1.395, P >0.05, compared to the healthy controls).In the surgery therapy cases, the IL-17 levels were also reduced after the thymus removal ( pre-surgery (8.8 ±1.4) pg/ml, 1 week after surgery (5.3 ±0.7) pg/ml, t=1.950, P<0.05;1 month after surgery (3.0 ±0.4) pg/ml, t=2.683, P<0.01).In the thymus tissues of the MG patients, the mRNA levels of IL-17R were much higher than that of the controls ( relative level 2.31 folds, t =2.682, P <0.01).Meanwhile, a positive correlation was found between the plasma IL-17 levels and the relative IL-17R levels in thymus tissues ( r =0.945 4, P <0.01 ).Furthermore, IL-17 was positively correlated with quantitative myasthenia gravis scores (QMGS) either pre-treatment (r =0.798 1, P <0.01) or post-treatment (r=0.906 5, P<0.01).And IL-17R was positively correlated with QMGS pre-treatment (r=0.775 5, P<0.01).Conclusions IL-17 is increased in the plasma of MG patients (both ocular and generalized) , and is decreased upon the glucocorticoid treatment or surgery therapy, suggesting that it can be used as a parameter to determine the therapeutic effects.IL-17R is increased in the thymus tissues of MG patients, suggesting that it can potentially be used as a pathological diagnosis parameter.

Objective:To explore the risk factors of intensive care unit-acquired weakness (ICU-AW) and the characteristics of Medical Research Council (MRC) score and electromyogram.Methods:A case control study was conducted. Patients with mechanical ventilation ≥ 7 days and MRC score admitted to department of respiratory and critical care medicine of China-Japan Friendship Hospital from September 2018 to January 2020 were enrolled, and they were divided into ICU-AW group (MRC score < 48) and non-ICU-AW group (MRC score ≥ 48) according to MRC score. The general situation, past medical history, related risk factors, MRC score, respiratory support mode, laboratory examination results, electromyogram examination results, ICU-AW related treatment, outcome and length of ICU stay were collected, and the differences between the two groups were compared. The risk factors of ICU-AW were analyzed by binary multivariate Logistic regression, and the characteristics of MRC score and electromyogram were analyzed.Results:A total of 60 patients were enrolled in the analysis, including 17 patients in ICU-AW group and 43 patients in non-ICU-AW group. Univariate analysis showed that there were significant differences in acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score, brain natriuretic peptide (BNP), blood urea nitrogen (BUN) on the first day of ICU admission and the ratio of invasive mechanical ventilation between ICU-AW group and non-ICU-AW group [APACHEⅡ score: 21 (18, 25) vs. 18 (15, 22), SOFA score: 7 (5, 12) vs. 5 (3, 8), BNP (ng/L): 364.3 (210.1, 551.2) vs. 160.1 (66.8, 357.8), BUN (mmol/L): 9.9 (6.2, 17.0) vs. 6.0 (4.8, 9.8), invasive mechanical ventilation ratio: 88.2% vs. 46.5%, all P < 0.05]. Binary multivariate Logistic regression analysis showed no independent risk factor for ICU-AW. The average MRC score of 17 ICU-AW patients was 33±11. The limb weakness was symmetrical, and the proximal limb weakness was the main manifestation. Electromyography examination showed that the results of nerve conduction examination in ICU-AW patients mainly revealed that the amplitude of compound muscle action potential (CMAP) and sensory nerve action potentials (SNAP) were decreased, and the conduction velocity was slowed down; needle electromyography showed increased area of motor unit potential (MUP), prolonged time limit and a large number of spontaneous potentials. Prognosis evaluation showed that compared with non-ICU-AW group, patients in ICU-AW group underwent more tracheotomy (70.6% vs. 11.6%), longer length of ICU stay (days: 57±52 vs. 16±8), and more rehabilitation treatment (58.8% vs. 14.0%), and the differences were statistically significant (all P < 0.01). Conclusions:The occurrence of ICU-AW may be related to high APACHEⅡ score and SOFA score, high levels of BNP and BUN on the first day of ICU admission and the proportion of invasive mechanical ventilation, but the above factors are not independent risk factors for ICU-AW. The MRC score of ICU-AW patients was characterized by symmetrical limb weakness, mainly proximal limb weakness; in electromyography examination, the nerve conduction examination results mainly showed that CMAP and SNAP amplitude were decreased, and conduction velocity was slowed down; needle electromyography examination showed increased MUP area, prolonged duration and a large number of spontaneous potentials.

Objective:To study the effect of Pterostilbene on endoplasmic reticulum stress and apoptosis in human renal tubular epithelial cells (HK-2 cells) induced by oxalate.Methods:From January 2019 to January 2020, HK-2 cells were divided into a control group (cultured with normal medium), an oxalate group (cultured with a medium containing 4 mmol/L of oxalate), and an intervention group of Pterostilbene (containing 4 mmol/L of oxalate + Pterostilbene 5, 10, and 20 μmol/L mixed medium were cultured at the same time), and the following tests were performed after 12 hours of treatment. Pterostilbene (5, 10, and 20 μmol/L) intervention group for cell viability test, lactate dehydrogenase cytotoxicity test, reduced glutathione, superoxide dismutase, malondialdehyde, hydrogen peroxide enzyme, total antioxidant capacity detection experiments to explore the degree of oxidative damage, and Western blotting experiments to explore the protein expression of ATF6, GRP78, DDIT3, caspase12, Clevead caspase 3/9; Pterostilbene (10 μmol/L) intervention group to detect mitochondrial membrane potential, caspase 3 enzyme activity, apoptosis rate, reactive oxygen detection to detect the apoptosis, reactive oxygen level, and qRT-PCR to detect ATF6, GRP78, DDIT3 of cells mRNA expression.Results:CCK-8 and lactate dehydrogenase toxicity test results showed that the cell activity of the oxalate group was significantly lower than that of the control group [(45.6±3.1)% vs. 100.0%, P<0.001]; the lactate dehydrogenase [(330.2±11.1)U/L vs. (2.6±6.7) U/L, P<0.001] of the oxalate group was higher than that of the control group increased obviously; the cell viability[ (57.2±1.7)%, (67.2±3.4)%, (78.9±1.8)%] of Pterostilbene intervention group (5, 10, 20 μmol/L) significantly increased compared with oxalate group ( P<0.05); lactate dehydrogenase [(288.1±4.3)U/L, (260.9±5.5)U, (202.7±10.2)U/L] in Pterostilbene intervention group (5, 10, 20 μmol/L ) was significantly lower than oxalate group ( P<0.05). The results of the five biochemical indexes of malondialdehyde, reduced glutathione, total superoxide dismutase, catalase, and total antioxidant capacity showed that the cell damage state was consistent with the experimental results of CCK-8 and lactate dehydrogenase. The active oxygen test results showed that the oxalate group had a significantly higher active oxygen level (76.3±4.9 vs. 6.2±1.7, P<0.01); the active oxygen level (39.5±5.4) of the Pterostilbene intervention group(10 μmol/L) was significantly lower than oxalate group ( P<0.01). The flow cytometry and caspase3 enzyme activity showed an increase in apoptosis rate and caspase3 activity in line with the trend of reactive oxygen levels. Mitochondrial membrane potential results showed that the oxalate group had a significantly lower mitochondrial membrane potential (0.76±0.15 vs. 7.84±0.26, P<0.01), and the mitochondrial membrane potential (2.26±0.27) of the Pterostilbene intervention group (10 μmol/L) was significantly higher than oxalate group( P<0.01). Western blot analysis showed that the relative expression of ATF6, DDIT3, GRP78, caspase12 and Cleaved caspase3/9 protein in the oxalate group was significantly higher than that in the control group. The relative expression of ATF6, DDIT3, GRP78, caspase12, Cleaved caspase3/9 protein in the Pterostilbene intervention group was significantly lower than that in the oxalate group ( P<0.05). qRT-PCR results showed that the mRNA expression trends of ATF6, DDIT3 and GRP78 in the three groups were consistent with the results of Western blotting. Conclusion:Pterostilbene can effectively inhibit the endoplasmic reticulum stress and apoptosis of HK-2 cells induced by oxalate.

Objective: To determine the effect of transplanting bone marrow mononuclear cells (BMMCs) on the expression of glial fibrillary acidic protein (GFAP) and Nogo-A around the ischemic foci after focal cerebral ischemia and reperfusion, and to study any role of BMMCs in nerve function recovery. Methods: BMMCs were isolated from the bone marrow of Sprague-Dawley rats. Cerebral ischemia and reperfusion was performed using a nylon thread to occlude the right middle cerebral artery for 2h followed by 24h of reperfusion. The qualified models were selected according to the Longa scale. The 48 models selected were randomly divided into a model group and an observation group, each of 24. Each group was further divided into 7d, 14d and 21d subgroups. 100μl of BMMCs (5×10⁶ /ml) were slowly injected into the ischemic lateral striata of the observation group. The rats in the model group were similarly injected, but with buffered saline solution. The rats were evaluated using the Longa scale after 7d, 14d and 21d. The rats were then sacrificed and the brain was resected. Immunohistochemical assays quantified the expression of GFAP and Nogo-A around the ischemic foci. Results: Compared with the model group, the rats in the observation group showed less neurological deficit on the 21st day, significantly greater expression of GFAP and significantly less Nogo-A expression on days 14 and 21. Nogo-A expression on the 21st day was also significantly lower than on the 14th day in the observation group. Conclusion BMMC transplantation can promote recovery from nerve damage after focal cerebral ischemia, which is probably related to enhanced expression of GFAP and restrained expression of Nogo-A in the brain tissues surrounding ischemic lesions.

Extracorporeal membrane oxygenation (ECMO) is a viable ultimate support therapy for patients with severe cardiorespiratory failure. Antimicrobial agents are commonly prescribed in critically ill patients with ECMO. However, an increasing number of studies have shown that ECMO circuit is associated with significant pharmacokinetic (PK) alterations, including the increased volume of distribution and reduced the clearance. In addition, the critical illness pathophysiology can also influence PK of antimicrobial, such as systemic inflammation, excessive fluid resuscitation, hypo-albuminemia, worsening hepatic or renal function. These PK alterations may increase the possibility of therapeutic failure or toxicity. Therefore, this study reviews published studies of the effects of ECMO on PK of antimicrobial agents in adults and makes preliminary recommendations on possible dosing regimen.

Background The ongoing new coronavirus pneumonia (Corona Virus Disease 2019,COVID-19) outbreak is spreading in China, but it has not yet reached its peak. Five million people emigrated from Wuhan before lockdown, potentially representing a source of virus infection. Determining case distribution and its correlation with population emigration from Wuhan in the early stage of the epidemic is of great importance for early warning and for the prevention of future outbreaks. Methods The official case report on the COVID-19 epidemic was collected as of January 30, 2020. Time and location information on COVID-19 cases was extracted and analyzed using ArcGIS and WinBUGS software. Data on population migration from Wuhan City and Hubei province were extracted from Baidu Qianxi, and their correlation with the number of cases was analyzed. Results The COVID-19 confirmed and death cases in Hubei province accounted for 59.91% (5806/9692) and 95.77% (204/213) of the total cases in China respectively. Hot spot provinces included Sichuan and Yunnan, which are adjacent to Hubei. The time risk of Hubei province on the following day was 1.960 times that on the previous day. The number of cases in some cities was relatively low, but the time risk appeared to be continuously rising. The correlation coefficient between the provincial number of cases and emigration from Wuhan was up to 0.943. The lockdown of 17 cities in Hubei province and the implementation of nationwide control measures efficiently prevented an exponential growth in the number of cases. Conclusion The population that emigrated from Wuhan was the main infection source in other cities and provinces. Some cities with a low number of cases showed a rapid increase in case load. Owing to the upcoming Spring Festival return wave, understanding the risk trends in different regions is crucial to ensure preparedness at both the individual and organization levels and to prevent new outbreaks.

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.
Chromatography, Liquid ; Cloning, Molecular ; Escherichia coli ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; Tandem Mass Spectrometry