OBJECTIVE:To establish an HPLC method for quantitative determination of baicalin in Yanyan capsules. METHODS:The determination was performed on Kromasil C18(250 mm?4.6 mm,5 ?m) with mobile phase consisted of meth-anol-water-phosphoric acid(47∶53∶0.2).The detection wavelength was set at 280 nm. RESULTS:The linear range of baicalin was 0.125～1.216 ?g(r=0.999 6) and its average recovery was 99.15%(RSD=0.62%,n=6).CONCLUSION:The method is simple and accurate,and it can be used for the quality control of Yanyan capsule.

Objective To observe the effect of fluoxetine on the single prolonged stress model which mimic the post-traumatic stress disorder (PTSD). Methods Rats receiving single prolonged stress (SPS) (2 h restraint + 20 min FST + anaesthesized to lose consciousness with ethylether) or not were given fluoxetine or tap water for 15 days. Elevated plus maze(EPM),open-field test(OF) and morris water maze(MWM) tests were used to evaluate rats' fear response to environment,high alertness,anxiety & depression behavior,and learning and memory ability. Results In open field test, group of fluoxetine(F1 (8895. 85 ± 599. 78) mm, (40. 23 ±4. 32) s;F2 (8654.07 ±866.05)mm,(41.57 ±4.34)s, P<0.05) showed significant increase in activity times and horizontal motion distance compared with group of SPS (4678.85 ±495.33)mm, (22.15 ±3.43)s, P<0.05). In EPM experiment,group of fluoxetine(F1 (32. 62 ± 4. 57)% , (17. 58 ± 3. 23)% ; F2 (39. 75 ± 4. 46)% , (19. 74 ± 4.44) %) showed significant increase in percentage of the open-arm into the maze and percentage of the open arm pause compared with group of SPS ((23.67 ±2. 87)% ,(12.46 ±2.55)% , P<0.05). In MWM experiment,the escape latency of the SPS group increased significantly in comparison to that in sham group (P<0.01) and fluoxetine group. Fluoxetine significantly reversed the SPS-induced decrease in time spent in the target quadrant (P< 0.05). Conclusion Added fluoxetine can obviously improve rats' fear response to environment ,high alertness ,anxiety & depression behavior as well as learning and memory ability.

Objective To investigate the clinical diagnostic value and pathogenesis of serum protein identification in Keshan disease (KD).Methods A total of 65 chronic KD patients were selected as the patient group in KD endemic areas,while 29 cases of dilated cardiomyopathy (the DCM group),62 healthy cases from KD endemic areas (control 1 group) and 28 healthy cases from non-endemic areas (control 2 group) were selected as controls.Liquid chip time of flight mass spectrometry (ClinProtTM MALDI-TOF-MS) was used to determine the expression of proteins/peptide peaks.ClinProTools 2.2 software was used to analyze the protein profiles to determine differentially expressed proteins/peptide peaks.The Genetic Algorithm (GA),QuickClassifer Algorithm (QC) and Supervised Neural Network Algorithm (SNN) methods were used to screen marker proteins.Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry technique (MALDI-TOF/TOF) was also used as a secondary mass spectrometry to identify differentially expressed peptides.Results Between the KD and control 1 groups,34 differentially expressed proteins/peptides and 5 marker proteins were identified,while 52 differentially expressed proteins/peptides and 5 marker proteins were identified between the KD and control 2 groups,and there were 67 differentially expressed proteins/peptides and 5 marker proteins between the KD and DCM groups.During secondary mass spectrometry,two peptides for mass-to-charge ratio (m/z) 2 079 and 1 465 were obtained,peptide of matching β-globin showed low expression while peptide of matching fibrinogen showed high expression in the KD patients.Conclusions Serum marker proteins can be used as biomarkers for diagnosis and differentiation of KD.β-globin and fibrinogen play an important role in the development of KD myocardial injury.

OBJECTIVETo investigate the frequency of p16 and p15 gene methylation in multiple myeloma (MM), and its relationship with bone marrow cell apoptosis and clinical outcome.

METHODSTwenty-two patients with MM were studied to detect p16 and p15 gene methylation. Methylation-specific polymerase chain reaction (MSP) was used to detect gene methylation, and terminal transferase-mediated dUTP nick end-labeling (TUNEL) was used to detect cell apoptosis.

RESULTSp16 and/or p15 gene methylatoin was detected in 10 of 22 patients (45.4%). There were 3 patients with p16 gene methylation, 9 patients with p15 gene methylation, and 2 patients with both genes methylation. The incidence of methylation of p15 gene was higher than that of p16 gene (P < 0.05). The patients with p16 and/or p15 gene methylation had a delayed cell apoptosis, poor response to chemotherapy, and a short over-all survival (OS).

CONCLUSIONThe methylation of p16 and/or p15 gene plays a key role in MM apoptosis pathogenesis. The patients with both p16 and p15 gene methylation had a poor prognosis.

Apoptosis ; Cell Cycle Proteins ; Cyclin-Dependent Kinase Inhibitor p15 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; DNA Methylation ; DNA, Neoplasm ; genetics ; Gene Silencing ; Genes, p16 ; Humans ; Multiple Myeloma ; genetics ; pathology ; Prognosis ; Transcription Factors ; genetics ; Tumor Suppressor Proteins

OBJECTIVETo scrutinize the immunohistochemical spectrum to differentiate hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma (ICC) and metastatic adenocarcinoma (MAC) in the liver.

METHODSSeven antibodies including AFP, Hep Par 1, CK18, CK19, CA19-9, CD34 and pCEA were immunohistochemically stained in resected specimens of 300 HCC, 35 ICC and 30 MAC. The specificity and sensitivity of the antibodies were evaluated by comprehensive capability score (CCS), with only those with CCS > or = 8 considered as having highly diagnostic value.

RESULTSAntibodies CCS > or = 8 were observed as Hep Par 1 and CD34 in HCC, and CK19 in ICC, but none in MAC. For HCC, CCS of Hep Par 1 was higher than that of AFP (9 vs. 7) with 83.7% in sensitivity and 96.7% in specificity.

CONCLUSIONFor HCC, Hep Par 1 and CD34 can be used as the first line antibodies, AFP and pCEA as the second line ones. CK19 is the first line antibody for ICC, and CA19-9 as the second. Hep Par 1, CD34 and CK19 are definitely helpful for the routine immunohistochemical stain to differentiate HCC from ICC and MAC.

Adenocarcinoma ; chemistry ; Antibodies ; immunology ; Carcinoma, Hepatocellular ; chemistry ; Cholangiocarcinoma ; chemistry ; Humans ; Immunohistochemistry ; Liver Neoplasms ; chemistry ; Tumor Cells, Cultured

Objective To explore the predictive value of early changes in platelet counts in the prognosis of severe pneumonia in aged patients.Methods This retrospective study included elderly patients with severe pneumonia,who were ≥65 years old and whose length of ICU stay ≥72 hours,admitted to the intensive care unit(ICU)of NO.2 People's Hospital of Changzhou from January 2014 to January 2017.They were divided into a survival group and a death group according to the 28-day outcome.General information and serum platelet levels at 0,24,36,and 72 hours after admission were collected.Receiver operating characteristic curve (ROC)was plotted according to platelet counts,changes in platelet counts and rates of change in platelet counts to evaluate their predictive value for 28-day prognosis.Kaplan-Meier survival curve was used to analyze the 28-day cumulative survival rate between different groups of patients,who were further divided according to platelet counts at 0 and 72 hours after admission to ICU,changes in platelet counts and rates of change in platelet counts at 72 hours after admission.Results (1)One hundred elderly patients with severe pneumonia were enrolled,among whom 41 cases were in the death group,thus with a mortality of 41.0％.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),sequential organ failure assessment (SOFA),C-reactive protein (CRP),and procalcitonin(PCT)in the death group were significantly higher than those in the survival group.2)Serum platelet levels showed a downward trend in both the death group and the survival group.The level of serum platelets at 72 hours after admission to ICU in the death group was significantly lower than that in the survival group(80.00 × 109/L vs.171.00 × 109/L,Z=5.786,P＜0.05);changes in platelet counts and rates of change in platelet counts in the death group were significantly higher than those in the survival group,especially at 72 hours after admission to ICU(/△PLT72:-79.00 × 109/L vs.-38.00 × 109/L,Z =4.938,P ＜0.05;△PLT 72％:43.6％ vs.-17.7％,Z=6.816,P＜0.05).(3)ROC curve analysis showed that platelet levels,changes in platelet counts,and rates of change in platelet counts could predict 28-day mortality in aged patients with severe pneumonia.The largest area under ROC curve was 0.902 when plotted with the rate of platelet counts at 72 hours after admission to ICU.Using the cut-off value of-36.14％ in the rate of change at 72 hours after admission to evaluate the predictive value in 28 day mortality,the sensitivity and specificity were 89.8％ and 75.6％,respectively.(4)Kaplan-Meier survival analysis showed that the 28 day survival rate was significantly higher and the length of survival was significantly greater when platelet counts at 0 and 72 hours after admission to ICU were higher than the cut-off value,and this also occurred in changes in platelet count and rates of change at 72 hours after admission to ICU.Conclusions Continuous decline in serum p[atelet levels indicates poor prognosis.When combined with platelet counts,changes in platelet counts and rates of change in platelet counts at 72 hours after admission to ICU,it may play an important role in assessing the prognosis of aged patients with severe pneumonia.

Medical ethics has a long history and rich connotations.It has developed from the simple "medical morality" of ancient times to the modem medical ethics.The basic principles of medical ethics include autonomy,non-maleficence,beneficence,justice,and so on.Researchers often conduct clinical researches in the balance between achievements and ethical norms.Clinical researchers of surgery should have a deep understanding of medical ethical principles and strictly abide by medical ethics.Ethics committee should strictly perform their duties and play the role of inspection and supervision.Modem medical knowledges should be popularized throughout the society to make clinical research correctly understood.Adhering principles of ethics first,people orientation and cooperation practice,with patients' benefit as evaluation criteria,balance of surgical "Dao" and "Shu" can be achieved.

Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV. It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.
Genome, Viral ; Haplotypes ; Humans ; Mutation ; Open Reading Frames ; Phylogeny ; SARS Virus ; genetics

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence ; China ; Cluster Analysis ; Gene Components ; Genetic Variation ; Genome, Viral ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; genetics