Purpose To study the values of liver histopathological assessment grading score in differential diagnosis between biliary atresia ( BA) and infantile hepatitis ( IHS) . Methods Thirty four cases of BA and sixteen cases of IHS were analyzed retrospectively, which were diagnosed by biopsy. A hepatic histopathological assessment grading score was developed. This consisted of eight features such as cholestasis, hepatocellular damage, bile duct proliferation, portal edema, portal inflammation, portal fibrosis, extramedullary hemopoiesis and multinucleated giant hepatocytes. The total scores were 24 points. All the cases were assessed one by one. Results The total scores of BA were significantly higher than that of IHS (P<0. 001). The frequencies of bile duct proliferation, portal fibrosis and portal edema were significantly higher in BA than that in IHS group, while the frequency of multinucleated giant hepatocytes was significantly higher in IHS than that in BA group. Conclusions This scoring system is helpful in differentiating BA from IHS.

OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.

OBJECTIVE:To optimize the water extraction technology of Yinju jiedu oral liquid,and provide reference for the industrial production of the preparation. METHODS:According to the investigation of extraction time-extraction rate curves of chlo-rohenic acid of Yinju jiedu formula and extraction rate of chlorohenic acid in Lonicera japonica and other combined medicinal mate-rials in the formula,decoction methods and time of L. japonica were determined. Using the comprehensive scores of linarin,harpa-goside,(R,S)-epigoitrin,psoralen+angelicin contents and dry extraction yield as indexes,L9(34)orthogonal test was designed to detect the effects of adding water amount,decoction time times and optimize the extraction technology of the residues and other me-dicinal materials. Verification test was conducted. RESULTS:The optimal technology was L. japonica decocted first for 30 min with 8-fold water;the residues and other medicinal materials were decocted with 8-fold water for 3 times,1 h each time;combin-ing all the syrups. In verification test,the average contents of chlorohenic acid,linarin,harpagoside,(R,S)-epigoitrin,psoralen+angelicin were respectively 34.51,10.31,1.97,0.21,9.79 mg/g(RSD=1.24％,1.19％,1.40％,1.71％,1.28％,n=3);aver-age dry extraction yield was 25.4％(RSD=1.64％,n=3);average extraction rate of chlorohenic acid was 78.95％(RSD=1.24％,n=3). CONCLUSIONS:In the optimized water extraction technology,both the extraction rate of chlorohenic acid and contents of other ingredients are relatively high. The technology is stable and feasible.

Objective To evaluate the association between LDL-C and ischemic stroke in patients with nonvalvular atrial fibrillation (AF).Method A total of 2 470 patients with nonvalvular AF were included in the present study.The clinical data and laboratory examination results of the patients in the hospital were collected.The subjects were either divided into the ischemic stroke history (n =560),and non-ischemic stroke history groups (n =1 910),or divided into the low-middle risk (n =566) and high risk groups (n =1 904) based on CHA2 DS2-VASc score.Results There were significant differences in the proportion of Han,the ratio of gender,age,hemoglobin,hematocrit,ALT,serum uric acid,HDL-C and LDL-C between the patients with ischemic stroke history and without (all P ＜ 0.05).Similarly,there were significant differences in the proportion of Han,the ratio of gender,age,white blood cell count,hemoglobin,hematocrit,platelet count,ALT,albumin,TG and LDL-C between subjects in the low-middle risk group and those in the high risk group (all P ＜ 0.05).A logistical regression analysis showed that LDL-C was an independent risk factor for both the ischemic stroke history (OR 2.089,95％ CI 1.860-2.347,P ＜0.05),and future ischemic stroke risk (OR 1.270,95％ CI 1.079-1.494,P ＜ 0.05) in patients with nonvalvular AF.Conclusion LDL-C is associated with ischemic stroke in patients with nonvalvular AF,and it is also an independent risk factor for future ischemic stroke in these patients.

In order to enhance the alkaline polygalacturonate lyase (PGL) productivity by Pichia pastoris, we developed a constant cell concentration culture strategy by methanol feeding (called as CCCM culture) used in the continuous cultures. We controlled reasonable cell concentrations in the bioprocess by different strategies of methanol feeding. Using this CCCM culture with DCW 75 g/L, we significantly enhanced the PGL productivity (Qv) and the average specific enzyme production rate (Qx) of PGL to 6.11 U/(mL x h) and 81.5 U/(g x h), increased by 42.1% and 191.2% than the fed-batch culture with high cell density, respectively. The final PGL activity was 441.9 U/mL. Moreover, the extracellular protease concentration is 1.9 mg/L and the cell viability is more than 94% after 120 hour induction. The results show that this new strategy is advantageous in reducing proteolytic degradation and enhancing cell viability.
Cell Count ; Culture Media ; Culture Techniques ; methods ; Fermentation ; Pichia ; genetics ; metabolism ; Polysaccharide-Lyases ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

Objective To observe the value of dual-source CT energy spectral imaging in qualitative evaluation of metastatic lymph nodes in patients with gastric cancer.Methods Data of patients with pathologically diagnosed gastric cancer were collected,who underwent both dual-source CT abdomen plain scanning and energy spectrum double-phase enhanced scanning.The long diameter,short diameter,short length-diameter ratio,arterial phase and venous phase standardized iodine concentration of lymph nodes were measured and recorded.Energy spectrum attenuation curves were drawn,and the slopes were calculated.Parameters with statistical significant differences between metastatic and non-metastatic lymph nodes were used to plot ROC curves,and the thresholds for diagnosing metastatic lymph nodes of gastric cancer,and their diagnostic sensitivity and specificity were calculated.Results There were significant differences in short axis,short aspect ratio,normalized iodine concentration in arterial phase and venous phase between metastatic and non-metastatic lymph nodes in patients with gastric cancer (all P＜0.05),the area under the curve of ROC curve was 0.601 (P=0.006),0.881 (P＜0.001),0.865 (P＜0.001) and 0.834 (P＜0.001),respectively.When the short length to diameter ratio was 0.730,the sensitivity and specificity in diagnosis of metastatic lymph nodes of gastric cancer was 75.72％ and 93.64％.When standardized iodine concentration was 0.223 in arterial phase,the sensitivity and specificity in diagnosis of metastatic lymph nodes of gastric cancer was 69.81％ and 91.20％.When the standardized iodine concentration was 0.392 in the venous phase,the sensitivity and specificity in diagnosis of metastatic lymph nodes of gastric cancer was 78.33％ and 85.63％.Dual-phase energy spectrum curve of both gastric metastatic lymph nodes and non-lymph node decreased.Taken the arterial curve slope k=6.860 as the threshold,the sensitivity and specificity in diagnosis of metastatic lymph nodes was 83.24％ and 84.72％.When the threshold venous phase slope k was set at 6.660,the sensitivity and specificity in diagnosis of metastatic lymph nodes was 64.62％ and 97.70％,respectively.Combining arterial phase length-to-diameter ratio,arterial phase standardized iodine concentration and spectral curve slope,the sensitivity and specificity in diagnosing metastatic lymph nodes was 97.34 ％ and 87.83 ％,which was 80.13 ％ and 90.31％ when combining venous phase short length-diameter ratio,venous phase standardized iodine concentration and spectral curve slope.Conclusion The short-length-to-diameter ratio,arterial-venous phase standardized iodine concentration and arterial-venous phase slope of lymph nodes are the main parameters for evaluating metastatic lymph nodes in patients with gastric cancer.Combination of the above three parameters can obviously improve the sensitivity and specificity in diagnosis of metastatic lymph nodes in patients with gastric cancer.

Objective To study the effect of estrogen on proliferation of astrocytes in hippocampus of mice following middle cerebral artery occlusion(MCAO).Methods One hundred and eight Kunming mice were randomly divided into estrogen group(n=54)and saline group(n=54).The animals in two groups underwent right MCAO with tissue samples taken at 3,6,12,24,48and 72h after MCAO.The ischemic site was detected and the ischemic size was measured with TTC staining,the damage of neurons in hippocampus was assayed with HE staining,the expression of GFAP in hippocampal astrocytes was detected with immunohistochemical staining.Results The cerebral infarction size was significantly smaller in estrogen group than in saline group at different time points after MCAO(P＜0.05,P＜0.01)especially at 12hafter MCAO(31.50％±3.36％vs 54.50％±5.68％,P=0.019).The damage of hippocampal neurons aggregated with the prolonged ischemia time in two groups and was milder in estrogen group than in saline group at the same time points.The expression level of GFAP positive cells in bilateral hippocampal areas was higher when the ischemia time was prolonged and was significantly higher in ischemic hippocampus of estrogen group than in that of control group except at 6hin CA3ischemic area(P＜0.05).Conclusion Estrogen can protect mice against focal cerebral ischemia,stimulate the genesis of astrocyte synapses,alleviate neuronal damage after ischemia,and can thus reduce the size of cerebral infarction.

Objective:To investigate the predictive value of sputum heparin binding protein(HBP) in sepsis related acute respiratory distress syndrome(ARDS).Methods:This study was a prospective case-control study.A total of 134 children with sepsis who were admitted in PICU at Hunan Children′s Hospital from January 2020 to November 2021 were included, including 63 children who had completed fiberoptic bronchoscopy.The 63 children were divided into sepsis without ARDS group, sepsis with mild ARDS group, and sepsis with moderate to severe ARDS group according to the presence and severity of ARDS.Sputum was collected and HBP was detected in all children with sepsis when they were admitted to the hospital.The alveolar lavage fluid within 72 hours of admission was reserved for HBP.The levels of interleukin (IL)-6 and tumor necrosis factor (TNF)- α were detected, and the blood biochemistry, pulmonary imaging, pediatric critical case score and other data within 72 hours were collected.Results:(1) Among 63 children with fiberoptic bronchoscopy, 29 were in sepsis without ARDS group, 18 were in the sepsis with mild ARDS group, and 16 were in the sepsis with moderate to severe ARDS group.There was no significant difference in the pediatric critical case score and the location of primary infection focus among the three groups at admission.The primary infection focus was respiratory system in 36 cases, whose sputum HBP level was (42.1±9.8) ng/mL, and 27 children with other systems infection, whose sputum HBP level was (37.8±10.8) ng/mL, there was no significant difference between two groups ( t=1.65, P=0.104). (2) There were significant differences in sputum HBP, alveolar lavage fluid HBP, IL-6 and TNF-α levels among sepsis with mild ARDS group, sepsis with moderate and severe ARDS group and sepsis without ARDS group ( P<0.05). The sputum HBP of 34 children with sepsis combined with ARDS was positively correlated with alveolar lavage fluid HBP, IL-6, TNF-α levels and lung injury score, and negatively correlated with SpO 2/FiO 2 ( P<0.05). (3)Among the 34 children with sepsis combined with ARDS, the sputum HBP concentration of children with invasive ventilation was significantly higher than that of children with non-invasive ventilation ( P<0.05). The sputum HBP concentration in children with three or more organ damage was significantly higher than that of children with two or less organ damage ( P<0.05). The sputum HBP concentration of dead children was higher than that of surviving children ( P<0.05). (4) The area under curve of sputum HBP for predicting ARDS was 0.772 (95% CI: 0.655~0.889). When the cut-off point value of sputum HBP was 27.9 mU/L, whose sensitivity and specificity were 70.6% and 79.3%, respectively.The area under curve of sputum HBP for predicting moderate and severe ARDS was 0.793 (95% CI: 0.661~0.926). When the cut-off point value of sputum HBP was 51.55 mU/L, whose sensitivity and specificity were 81.3% and 76.6%, respectively. Conclusion:Sputum HBP is elevated in children with sepsis and ARDS, which is related with the severity of the disease.Sputum HBP has a good predictive value for the diagnosis and severity of children with sepsis and ARDS, and can be used as a clinically effective and convenient evaluation index for children with sepsis related ARDS.

Objective To investigate the efficacy of leflunomide combined with prednisone in the induction therapy of proliferative lupus nephritis (LN).Methods A prospective,multicenter,randomized controlled clinical trial was conducted in patients with biopsy-proved proliferative lupus nephritis recruited from 15 renal centers from 2013 to 2015.Patients were randomized to two groups.Oral leflunomide or intravenous cyclophosphamide was given to patients in each group.Both groups received a tapering course of oral prednisone therapy.All patients were followed up for 24 weeks.The blood biochemistry,urine index,clinical curative effect and adverse reaction were recorded and analyzed statistically.Results A total of 100 patients were enrolled in this clinical trial,including 48 patients in leflunomide group and 52 patients in cyclophosphamide group.After 24 weeks,the overall response rate was 79％ (95％ CI 67％-90％) in the leflunomide group and 69％ (95％ CI 56％-82％) in the cyclophosphamide group.23％ (95％CI 11％-35％) of patients in leflunomide group showed complete remission compared with 27％ (95％CI 24％-30％) in cyclophosphamide group (P=0.35).The levels of 24-hr urine protein excretion,SLEDAI and anti-dsDNA antibody titers were decreased in patients treated with leflunomide group after 24-weeks treatment.And the levels of serum albumin and complement 3 after treatment were significantly higher compared with these before treatment.There was also no significant difference in changes of 24-hr urine protein excretion,SLEDAI score,anti-dsDNA antibody titers,serum albumin and complement C3 levels after treatment between two groups.Incidence of adverse events did not differ between the leflunomide and cyclophosphamide group.Conclusions Leflunomide combined with prednisone showed same efficacy compared with cyclophosphamide as induction therapy for lupus nephritis.Leflunomide might be an useful medicine in the induction therapy of lupus nephritis.

Objective To analyze and summarize the cases of pregnancy on maintenance hemodialysis (MHD),and review the literature.Methods Seven cases of pregnancy and childbirth in patients on MHD in the First Affiliated Hospital of Zhejiang University from Jan 2009 to Dec 2017 were analyzed,and the literature about pregnancy in patients on MHD reported in Pubmed and Web of Science database was retrieved.Both maternal and fetal outcome were studied.Results There are seven pregnant MHD patients in this center,among whom six patients went through a smooth pregnancy and one patient had intra-uterine fetal death at 14 weeks of pregnancy.The six patients had preterm labor.Among six fetuses,four grew in good health and developed well,one had physical development retardation and one had heart malformation at born.In the literature,169 cases reported pregnant patients and 182 fetuses were evaluated,of which 145 live infants were delivered,79.67％ overall fetal survival rate,with gestational age of (32.94±3.34) weeks.In 164 delivered fetuses,147 were preterm labor (89.63％).The weight of live births was (1824±609) g.There were no maternal deaths.Fetal survival rate was zero in ＜ 20 weeks of gestational age,20-24 weeks was 14.29％,25-27 weeks was 45.45％,≥28 weeks was 94.44％.The fetal survival rate was higher in pregnant woman receiving hemodialysis≥28 hours weekly compared to women receiving ＜ 16 hours weekly (92.31％ vs 52.94％,P=0.02).Conclusions There are still a very high matemal and fetal complication rate in hemodialysis patients,especially in fetus.Gestational age≥28 weeks has a high fetal survival rate.Intensive dialysis during pregnancy may benefit higher fetal survival rate.