Objective To establish a HPLC method for the determination of Shionone in Ziwan Zhisou granule. Methods Phenomenex C18 column (250? 4.6 mm) was used. The mobile phase was acetonitrile and the detection wave length was at 203 nm. Result A good linearity of Shionone was in the range of 0.025～ 0.250 ? g/? L, r=0.9996. The average recovery was 97.8 % , RSD was 2.37 % ( n=5) . Conclusion This method is sensitive, accurate and simple and with good reproducibility for the determination of Shionone in Ziwan Zhisou granule. 

Objective:To analyze the risk factors and clinical characteristics of liver injury in patients with sepsis and to provide a reference for early recognition, early diagnosis, early intervention, and improve the survival rate of patients.Methods:The clinical data of sepsis patients admitted to the department of general intensive care unit (ICU) of the Second Affiliated Hospital of Zhejiang University School of Medicine from July 2014 to October 2020 were retrospectively analyzed. According to the occurrence of acute liver injury, patients with sepsis were divided into the liver injury group and the non-liver injury group, and the differences of demographic data, history, history of primary diseases, laboratory indicators on the first time of admission, treatments, the severity of the disease and other indicators were compared and analyzed. Logistic regression was used to analyze the risk factors for sepsis-related liver injury.Results:A total of 527 patients with sepsis were enrolled, and 129 patients with acute liver injury, accounting for 24.48%. Compared with the non-liver injury group, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), sequential organ failure assessment (SOFA), pro-brain natriuretic peptide (pro-BNP), serum MB isoenzyme of creatine kinase (CK-MB), total bile acid (TBA), serum creatinine (SCr), blood urea nitrogen (BUN), lactic acid (Lac), lactate dehydrogenase (LDH), C-reactive protein (CRP), procalcitonin (PCT) in liver injury group were significantly increased [APACHEⅡ score: 23.00±10.40 vs. 16.10±8.10, SOFA score: 9.17±4.29 vs. 5.90±3.12, pro-BNP (ng/L): 5 500.0 (1 166.0, 16 865.0) vs. 1 377.2 (448.8, 6 136.5), CK-MB (U/L): 23.0 (13.0, 55.0) vs. 18.0 (13.0, 31.0), TBA (μmol/L): 5.0 (2.4, 12.9) vs. 2.6 (1.4, 4.9), SCr (μmol/L): 146.0 (75.0, 222.0) vs. 71.0 (52.0, 125.8), BUN (mmol/L): 13.4 (8.8, 20.2) vs. 7.9 (4.9, 11.6), Lac (mmol/L): 2.0 (1.4, 4.4) vs. 1.4 (1.0, 2.2), LDH (μmol·s -1·L -1): 6.43 (3.76, 11.99) vs. 4.55 (3.38, 6.63), CRP (mg/L): 113.0 (61.8, 201.0) vs. 95.0 (37.3, 170.1), PCT (μg/L): 3.8 (1.0, 23.3) vs. 0.8 (0.2, 6.4)], prothrombin time (PT), international standard ratio (INR) and activated partial thrombin time (APTT) were significantly longer [PT (s): 19.4±7.6 vs. 16.0±4.0, INR: 1.7±1.0 vs. 1.3±0.5, APTT (s): 54.0±25.8 vs. 44.1±15.1], plasma fibrinogen (FIB), platelet count (PLT), albumin (ALB), and cholesterol (CHOL) were decreased [FIB (g/L): 4.2±2.3 vs. 4.9±1.8, PLT (×10 9/L): 116.3±74.3 vs. 182.7±108.6, ALB (g/L): 25.4±5.5 vs. 27.6±5.5, CHOL (mmol/L): 2.5±1.2 vs. 3.2±1.3], the probability of shock was significantly increased (91.47% vs. 59.19%), and the duration of shock was prolonged [days: 5.0 (2.0, 9.0) vs. 1.0 (0.0, 3.0)], positive rate of microbial culture (81.40% vs. 71.11%), probability of occurrence of drug-resistant bacteria (67.44% vs. 47.99%) were significantly higher, mechanical ventilation time [days: 6.0 (2.0, 12.7) vs. 2.4 (0.0, 6.9)], continuous renal replacement therapy (CRRT) time [days: 1.2 (0.0, 5.0) vs. 0.0 (0.0, 0.0)], the length of intensive care unit (ICU) stay [days: 9.0 (5.0, 18.0) vs. 7.0 (3.0, 13.0)] were significantly longer, 28-day mortality was significantly higher (80.62% vs. 28.89%), and the differences were statistically significant (all P < 0.05). Further Logistic regression analysis showed that PLT decline, PT prolongation, CRRT duration, shock duration and 28-day mortality were correlated with sepsis-related liver injury [odds ratios ( OR) and 95% confidence interval (95% CI) were 0.992 (0.987-0.998), 3.103 (1.507-6.387), 1.198 (1.074-1.336), 1.196 (1.049-1.362), and 0.213 (0.072-0.633), respectively, all P < 0.05]. Conclusions:Prolonged PT and decreased PLT are independent risk factors for sepsis complicated with liver injury. The long duration of CRRT, long duration of shock, and high mortality are independent clinical characteristics of patients with sepsis-related liver injury.

BACKGROUND:Posterior stabilized femoral knee prosthesis needs additional condyle osteotomy to accommodate the tibial post and femur fossa structures. Intercondylar fossa on both sides connected at the femoral body with concentrated stress is a place easily affecting fractures. Differences in bone mass between different models of different brands did not have specific data, which was not convenient to select prosthesis for clinicians.
OBJECTIVE: To compare the difference of intercondylar osteotomy data among clinical commonly used posterior stabilized knee prostheses (six imported and domestic brands), and to provide basis for the selection and application of the prostheses.
METHODS:The current commonly used posterior stabilized knee prostheses (six imported and domestic brands) were used, including Zimmer NexGen LPS, Stryker Scrorpio NRG Knee-Flexed, Depuy PFC Sigma, Smith & nephew Genesis-2 PS, United-U1 and Wego GKPS. According to the osteotomy template, the osteotomy-surfaces consisting of femoral condyle starting section and cross section, distal section of femoral condyle, and back-oblique section were identified. The corresponding femoral prosthesis diameter lines included condylar ambilateral and anteroposterior diameters, width and depth of femoral intercondylar fossa. The above data were compared and measured.
RESULTS AND CONCLUSION:The six kinds of knee femoral prostheses were different in ratio of ambilateral diameter and anteroposterior diameter, bone resection of intercondylar fossa, and geometry. Imported prostheses carry shorter diameters in femoral starting and cross sections, so it can catch more posterior condylar osteotomy. With increasing prosthesis sizes, the ratio of bone loss causing by width of intercondylar osteotomy is decreased among six brands. In al sizes, Stryker Scrorpio NRG Knee-Flexed catches shorter width of intercondylar osteotomy. Knee prosthesis osteotomy among six brands is different. The result of this study is not sufficient to evaluate the pros and cons between different prostheses, but as reserving bone is concerned, the design of less intercondylar osteoomy catches more advantages.

Objective: To observe hypoglycemic effects of Yunu Decoction, Zuogui Pill and Shenqi Pill, three compound traditional Chinese herbal medicines, in treatment of diabetes mellitus induced by alloxan in rats, and to compare the therapeutic effects among the three recipes for nourishing yin, clearing away heat, and nourishing yin and warming yang. Methods: Diabetes mellitus was induced in rats with alloxan at a dose of 60 mg/kg via tail vain injection. The diabetic rats were randomly divided into four groups: alloxan model group, Yunu Decoction-treated group, Zuogui Pill-treated group and Shenqi Pill-treated group. Rats in the three recipe groups were administered intragastrically with water extraction of Yunu Decoction, Zuogui Pill, and Shenqi Pill accordingly for 10 days. Then the level of blood glucose was measured by glucose oxidase method and the glucose tolerance was determined. Results: Compared with the normal rats, blood glucose level in the alloxan model group was obviously increased (P<0.05). Glucose levels in the three recipe groups were obviously decreased as compared with the alloxan model group (P<0.05), and glucose level in the Yunu Decoction-treated group after treatment was significant lower than before treatment (P<0.05). The glucose tolerance test indicated that rats in the alloxan model and three recipe groups revealed impaired glucose tolerance as compared with the normal rats, and there were no significant differences between the alloxan model group and the three recipe groups. Conclusion: Yunu Decoction, Zuogui Pill and Shenqi Pill can effectively decrease the glucose level of the rats with diabetes mellitus induced by alloxan, and Yunu Decoction showed the best therapeutic effects. The glucose tolerance test shows that the three recipes cannot correct the abnormal metabolism of glucose.

Objective To observe the intervention effect of immune-enhancing enteral nutrition (EN) emulsion on immune function of critically ill patients with mechanical ventilation (MV). Methods One hundred and twenty critically ill patients with MV admitted to the Department of Emergency Intensive Care Unit (EICU) of Taizhou First People's Hospital from July 2015 to June 2017 were enrolled, and they were divided into immune-enhancing EN group and standard EN group by random numbers generated by a computer. Ultimately, 76 cases were enrolled in the study, among them, 36 cases were in the immune-enhancing nutrition group and 40 cases were in the standard nutrition group. The differences of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), the sequential organ function evaluation (SOFA) score on 1, 3, 7 days and immunity indexes (secretory immuno-globulins IgA, IgG, IgM), lymphocyte subpopulation (CD4 and CD8), duration of MV and the length of ICU stay on the 1, 7 days after EN were compared. Results Comparisons between the immune-enhancing EN group and standard EN group showed: APACHE Ⅱ score had no statistically significant difference between the two groups at each time point, SOFA score on 7 days after EN treatment was significantly decreased in the immune-enhancing EN group (2.56±1.38 vs. 3.68±2.96, P < 0.05); IgA, IgG, IgM were obviously higher in the immune-enhancing EN group than those in standard EN group on 7 days after treatment [IgA (mg/L): 2 967.6±635.6 vs. 2 525.0±592.7, IgG (mg/L): 14 982.5±2 899.7 vs. 12 996.4±2 875.9, IgM (mg/L): 1 206.8±233.3 vs. 1 093.2±165.1, all P < 0.05], CD4 (0.45±0.06 vs 0.37± 0.10) and CD8 (0.20±0.03 vs. 0.18±0.04) were significantly higher than those in standard EN group (both P < 0.05). The MV time (hours): 122.33±63.91 vs. 155.69±77.06) and ICU stay time (hours): 197.57±70.60 vs. 239.61±84.83) of the immuno-enhancing EN group were markedly shorter than those of the standard EN group (both P < 0.05). Conclusion Compared with standard EN, the immune-enhancing EN emulsion can improve the immune function of critically ill patients with MV, and shorten the duration of MV support and the length of ICU stay.

Objective To investigate the role of enteral nutrition (EN) support in the treatment of patients with chronic heart failure. Methods Ninety patients with chronic heart failure (conform to the New York Heart Association (NYHA) cardiac function class Ⅲ-Ⅳ) admitted to Intensive Care Unit (ICU), Cardiology Care Unit (CCU) and Emergency ICU (EICU) of Taizhou First People's Hospital from January 2015 to September 2017 were enrolled, and according to different nutritional methods, they were divided into a control group (rational autonomous diet group) and an observation group (Ruineng enteral nutritional emulsion for EN group), each group 45 cases. Based on the calculation (Harris-Benedict) of individual total energy consumption the control group had a reasonable autonomous diet and Ruineng EN emulsion for EN group. The chang of various nutrition indexes [including body mass index (BMI), serum total protein (TP), albumin (Alb), hemoglobin (Hb), vitamin B12, folic acid, serum iron], inflammatory factors [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)], and the level of cardiac function index (LVEF) before and after treatment were observed. Results ① Before treatment, vitamin B12 in the observation group was significantly higher than that in the control group (ng/L: 153.3±54.6 vs. 113.4±80.2, P < 0.05), there were no statistical significant differences in other indicators between the two groups (all P > 0.05). ② After treatment, compared with those before treatment, the nutritional indicators and LVEF of both groups were higher, and inflammatory factors were lower, there were statistical significant differences in the other indicators before and after treatment except Hb and IL-6 in the control group and serum iron in the observation group [the control group: BMI (kg/m2) was 20.9±1.8 vs. 19.9±1.2, TP (g/L) was 66.0±2.4 vs. 63.7±1.6, Alb (g/L) was 34.4±3.5 vs. 31.1±2.3, vitamin B12 (ng/L) was 149.5±79.2 vs. 113.4±80.2, folic acid (nmol/L) was 10.0±1.7 vs. 4.6±3.2, serum iron (μmol/L) was 16.5±13.7 vs. 10.4±7.5, TNF-α (ng/L) was 23.8±10.0 vs. 28.3±8.6, LVEF was 0.35±0.14 vs 0.32±0.04; observation group: BMI (kg/m2) was 21.5±1.4 vs. 20.2±1.4, TP (g/L) was 66.5±2.8 vs. 64.3±2.2, Alb (g/L) was 35.8±3.1 vs. 33.3±1.9, Hb (g/L) was 121.4±13.8 vs. 112.9±12.0, vitamin B12 (ng/L) was 201.1±98.6 vs. 153.3±54.6, folic acid (nmol/L) was 15.7±14.4 vs. 8.8±2.8, TNF-α (ng/L) was 20.5±6.3 vs. 25.8±3.0, IL-6 (ng/L) was 209.4±6.5 vs. 220.9±16.9, LVEF was 0.38±0.07 vs. 0.33±0.02, all P < 0.05]. ③ Before and after treatment, the changes of BMI, Hb, vitamin B12, folic acid and IL-6 in the observation group were more significant than those in the control group [BMI (kg/m2): 1.4±0.9 vs. 1.1±0.3, Hb (g/L): 8.6±1.2 vs. 2.7±0.9, vitamin B12 (ng/L): 47.1±1.0 vs. 36.2±0.9, folic acid (nmol/L): 6.8±1.8 vs. 5.5±1.8, IL-6 (ng/L):-10.8±2.3 vs. -1.6±1.0, all P < 0.05]. After treatment, the degree of increase of serum iron in the control group was more significant than that in the observation group (μmol/L: 6.2±0.8 vs. 1.4±0.9, P <0.05), there were no significant differences in the degrees of improvement in TP, Alb and TNF-α between the two groups (all P > 0.05). ④ The difference value of each indicator before and after treatment of the two groups of patients with cardiac grade Ⅲ was more significant than that in the patients with cardiac grade Ⅳ, among the indicators in the control group, Hb, serum iron and IL-6 showed statistical significant differences [Hb (g/L): 3.05±0.42 vs. 2.47±0.84, serum iron (μmol/L): 6.81±0.91 vs. 5.95±1.82, IL-6 (ng/L): -3.87±0.45 vs. -0.53±0.28, all P < 0.05], while in the observation group of patients with cardiac grade Ⅲ and Ⅳ, Alb, Hb, serum iron, IL-6 appeared statistical significant differences [Alb (g/L): 3.41±0.38 vs. 2.27±0.91, Hb (g/L): 9.83±1.44 vs. 8.10±0.98, serum iron (μmol/L): 2.23±0.34 vs. 1.04±0.88, IL-6 (ng/L):-14.11±0.42 vs. -9.45±1.01, all P < 0.05]. Conclusion In the treatment of patients with chronic cardiac failure, simultaneously EN support is given energetically, that can improve the nutrition status of organism, reduce inflammatory reaction and enhance cardiac function; the therapeutic effect of Ruineng EN support is remarkably better than that of the autonomous diet support.

Objective To study the relationship of serum 25-hydroxy vitamin D [25-(OH) D] and vitamin D binding protein (DBP) in premature infants with bronchopulmonary dysplasia (BPD) and their clinical significance.Method From March 2017 to September 2018,the premature infants with gestational age (GA)＜32 weeks admitted to the neonatal department of our hospital were prospectively studied.All the premature infants were given 800 IU/d vitamin D supplement from one week after birth.Venous blood sample were collected at birth and 28 d after birth to measure 25-(OH) D aud DBP levels.The infants were evaluated for BPD at 28 d after birth and then assigned into the BPD group and the non-BPD group.The differences of 25-(OH) D and DBP levels were compared.Result A total of 170 premature infants (GA＜32 weeks) were included,including 56 cases in the BPD group and 114 cases in the non-BPD group.The BPD group had 34 males,the GA was (29.8±1.2) weeks,the birth weight (BW) was (1 198± 157) g.The non-BPD group had 95 males,the GA was (30.2± 1.5) weeks,the BW was (1 243± 146) g.No significant differences existed in GA,BW and male gender proportion between BPD group and non-BPD group (P＞0.05).The BPD group had a lower levels of serum 25-(OH) D at birth [(27.8±5.9) nmol/L vs.(30.4±1.1) nmol/L,P＜0.05].The levels of serum 25-(OH) D in moderate/severe BPD group were significantly lower than mild BPD group [(25.3±4.9) nmol/L vs.(29.7±5.9) nmol/L,P＜0.05];25-(OH) D in BPD group was still lower than the non-BPD group at 28 days after birth (after vitamin D supplement) [(77.5±11.7) nmol/L vs.(83.8±11.6) nmol/L,P＜0.05].Comparison of serum DBP levels between the two groups showed that,DBP at 28 d after birth in BPD group were significantly lower than the non-BPD group,and DBP in moderate/severe BPD group were significantly lower than the mild BPD group [(373.9± 19.1) μg/ml vs.(391.4±23.6) μg/ml],the differences were both statistically significant (P＜0.05).Multivariate analysis showed that the high serum 25-(OH)D level at birth (OR=0.827,95％CI0.693～0.987) was protective factors for BPD,while neonatal pneumonia (OR=4.331,95％CI 1.269～14.784) and neonatal sepsis (OR=4.020,95％CI 1.153～14.015) were risk factors for BPD.Conclusion The high serum 25-(OH) D level at birth in preterm infants was protective factors for BPD,while neonatal pneumonia and sepsis were the risk factors for BPD.Moreover,low serum 25-(OH) D level at birth and low serum DBP level at 28 d after birth maybe useful indicators for the severity of BPD.

Objective: To investigate the dynamic change of paraquant-induced kidney injury in rats and the protective effect of edaravone. Methods: Eighty SD rats were randomly divided into 4 groups: the normal control group, paraquat poisoning group, edaravone treatment group and edaravone control group. The normal control group of 8 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and the same amount of fluid into the abdominal cavity after 30 minutes. The paraquat poisoning group of 24 rats were given 1 ml of paraquat solution (20 mg/kg) through the abdominal cavity to build poisoning models, and the same amount of 0.9% sodium chloride was injected into the abdominal cavity after 30 minutes. The edaravone treatment group of 24 rats were given edaravone (5 mg/kg) through the abdominal cavity after 30 minutes when the poisoning models were set up. The edaravone control group of 24 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and edaravone (5 mg/kg) was injected into the abdominal cavity after 30 minutes. In addition to the normal control group, the other groups processed 1 times a day to mantain 7 d. On 1, 3, 7, 21 d several rats in each group were excuted and the kidney tissue and serum samples were collected, then each pathological changes of the kidney were observed with light microscopy. Serum creatinine, KIM-1, NGAL were measured by ELISA, the expression of HSP70 protein in kidney were observed with immunohistochemical staining. Results: The pathological examination reveald that the damage of kidney tissue in the paraquat group was the most serious on 3 d, and the damage was consistently alleviated in edaravone treatment group at the same time, renal fibrosisn was unseen in each group until 21 d. Compared with normal control group, there was no statistically significant in edaravone control group (P>0.05) . The KIM-1 in blood and kidney in paraquat poisoning group were markedly increased in 1 d (P<0.05) . The NGAL in blood and creatinine were markedly increased in d7 (P<0.05) . The NGAL in kidney increased over time, but had no statistically difference with the control group (P>0.05) .Compared with paraquat poisoning group, the serum creatinine, KIM-1 in blood and kidney, the KIM-1 in kidney had decreased significantly in edaravone treatment group (P<0.05) . The NGAL in kidney has no statistically significant compared with the poisoning group (P>0.05) . HSP70 expression of kidney tissue in edaravone treatment group had significantly increased in d3 compared with the paraquat poisoning group (P<0.05) . Conclusion Edaravone can prompt a significant rise of HSP70 in kidney tissue, reduce KIM-1 and NGAL levels, and play a protective role in kidney injury of acute paraquat poisoning.

OBJECTIVETo explore the possible mechanism and protective effect of BMSCs (bone mesenchymal stem cells) carrying superoxide dismutase (SOD) gene on mice with paraquat-induced acute lung injury.

METHODSTo establish the cell line of BMSCs bringing SOD gene, lentiviral vector bringing SOD gene was built and co-cultured with BMSCs. A total of 100 BALB/c mice were randomly divided into five groups, namely Control group, poisoning group (PQ group) , BMSCs therapy group (BMSC group) , BMSCs-Cherry therapy group (BMSC-Cherry group) , BMSCs-SOD therapy group (BMSC-SOD group) . PQ poisoning model was produced by stomach lavaged once with 1 ml of 25 mg/kg PQ solution, and the equal volume of normal saline (NS) was given to Control group mice instead of PQ. The corresponding BMSCs therapy cell lines were delivered to mice through the tail vein of mice 4h after PQ treatment.Five mice of each group were sacrificed 3 d, 7 d, 14 d and 21 days after corresponding BMSCs therapy cell lines administration, and lung tissues of mice were taken to make sections for histological analysis. The serum levels of glutathione (GSH) , malondialdehyde (MDA) , SOD, and the levels of transforming growth factor-β (TGF-β) and tumor necrosis factor-α (TNF-α) in lung tissue were determined. The level of SOD was assayed by Westen-blot.

RESULTSCompared with Control group, the early (3 days) levels of SOD protein in lung tissue of PQ group obviously decreased, and the late (21 days) levels of SOD obviously increased, while in therapy groups, that was higher than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Compared with Control group, the levels of plasma GSH and SOD of PQ group and each therapy group wae significantly lower than those in Control group, while in therapy groups, those were higher than those of PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) .Compared with Control group, the level of plasma MDA, TNF-α and TGF-β in PQ group and therapy groups were significantly higher, while in therapy groups, that was lower than that in PQ group, and the BMSCs-SOD group showed most obvious (all P<0.05) . Lung biopsy showed that, the degree of lung tissue damage in each therapy group obviously reduced.

CONCLUSIONSOD is the key factor of the removal of reactive oxygen species (ROS) in cells, that can obviously inhibit the oxidative stress damage and the apoptosis induced by PQ, thus significantly increasing alveolar epithelial cell ability to fight outside harmful environment.

Acute Lung Injury ; chemically induced ; therapy ; Animals ; Antioxidants ; metabolism ; Cell Line ; Glutathione ; blood ; Lung ; pathology ; Malondialdehyde ; blood ; Mesenchymal Stem Cell Transplantation ; Mice ; Mice, Inbred BALB C ; Oxidative Stress ; Paraquat ; poisoning ; Superoxide Dismutase ; blood ; genetics ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.