BACKGROUNDThrombin is a multifunctional serine protease that plays a crucial role in hemostasis following tissue injury. In addition to its procoagulation effect, thrombin is also a potent mesenchymal cell mitogen, therefore it plays important roles in the local proliferation of mesenchymal cells in the tissue repair process. Reactive oxygen species (ROS) can induce some human cells to proliferate at lower rates while at higher concentrations they promote cells to undergo apoptosis or necrosis. Accumulative evidence suggests that thrombin can induce some cells to produce ROS. Based on these observations, we provide a hypothesis that thrombin can stimulate human lung fibroblasts to produce ROS, which play an important role in human lung fibroblast proliferation.

METHODSROS were detected in fibroblasts at 30 minutes and 60 minutes following thrombin (20 U/ml) exposure using flow cytometry. The ratio of reduced glutathione/oxidized glutathione (GSH/GSSG) was assayed in lung fibroblasts using a commercial kit following treatment with thrombin at different concentrations. NADPH oxidase and the extracellular regulated kinase1/2 (ERK1/2) signaling pathway were detected by Western blotting after thrombin stimulation to lung fibroblasts.

RESULTSThrombin, at 20 U/ml, stimulated human lung fibroblasts (HLF) to generate ROS in a time dependent manner. The ratio of GSH/GSSG in fibroblasts treated with thrombin showed a significant decrease. NADPH oxidase was activated and the ERK1/2 signal pathway was involved in the proliferation process of fibroblasts treated with thrombin.

CONCLUSIONThe activation of NADPH oxidase by thrombin leads to the production of ROS, which promotes fibroblasts proliferation via activation of the ERK1/2 signaling pathway.

Cell Proliferation ; drug effects ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; analysis ; physiology ; Fibroblasts ; drug effects ; physiology ; Flow Cytometry ; Glutathione ; metabolism ; Humans ; Lung ; cytology ; NADPH Oxidases ; analysis ; physiology ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; physiology ; Thrombin ; pharmacology

OBJECTIVETo demonstrate molecular characterization of a newly isolated enterovirus.

METHODSVirus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5'3'RACE (rapid amplification of cDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus.

RESULTSSequence analysis showed that this newly isolated enterovirus shared 83%-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89. Phylogenetic analysis indicated that it was probably a new subtype of enterovirus 89.

CONCLUSIONThis newly isolated enterovirus in the stool specimen from patient has the same serotype with enterovirus 89, but it was probably a new subtype of enterovirus 89.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Enterovirus ; classification ; genetics ; isolation & purification ; Feces ; virology ; Genome, Viral ; genetics ; Humans ; RNA, Viral ; genetics ; Sequence Analysis, DNA

Objective To investigate the change of nursing undergraduates ’ attitude towards patients with mental disorder between before and after their nursing practice in the psychiatric department .Methods One hundred and fifteen students were selected and provided an internship program delivered by the Nursing Department to practice in the Psychiatric Hospital .All of nursing undergraduates were investigate with stigma scale towards people with mental disorder and the change of their attitude towards patients with mental disorder were compared between before and after their nursing practice .Results After nursing practice, the total score of the stigma scale among nursing undergraduates was (40.90 ±14.84), the score of dangerous dimension was (13.56 ±5.44), and the score of social isolation dimension was (22.09 ±9.55), which were all lower than those before nursing practice , these differences were statistically significant (t =3.86, 4.02, 3.02, respectively;P<0.01).The score of contact scale among nursing undergraduates after nursing practice was (3.64 ±1.29) and higher than that before practice with (2.96 ±0.67), the difference was statistically significant (t=-3.68;P<0.01).The nursing undergraduates ’ willingness to work in the psychiatric hospital increased to 71.3%after nursing practice from 49.6%before nursing practice , the difference was statistically significant (χ2 =230.00, P<0.01).Conclusions The clinical nursing practice for nursing undergraduates in the psychiatric department can reduce their discrimination against patients with mental disorder .

Objective To demonstrate molecular characterization of a newly isolated enterovirus.Methods Virus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5' ,3'RACE (rapid amplification of eDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus. Results Sequence analysis showed that this newly isolated enterovirus shared 83 %-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89.Phylogenetic analysis indicated that it was probably a new subtype of enterovirus 89. Conclusion This newly isolated enterovirus in the stool specimen from patient has the same serotype with enterovirus 89, but it was probably a new subtype of enterovirus 89.

Objective:To explore the adjunctive diagnostic value of transcranial sonography (TCS) combined with olfactory test in early Parkinson′s disease (PD) and the clinical value of both in the cognitive function of PD patients.Methods:TCS and olfactory test were performed in 157 early PD patients(PD group) and 157 healthy controls(control group) in the Second Affiliated Hospital of Soochow University from January 2018 to January 2022. The differences in clinical characteristics, TCS, and olfactory test results between the two groups were analyzed. The values of TCS, olfactory test, and their combination in diagnosing early PD were evaluated using clinical diagnosis as the gold standard. The correlations of the midbrain area, the midbrain substantia nigra hyperechoic area, and the third ventricle width in TCS examination with the cognitive score were analyzed in the PD group. According to the olfactory test scores, 157 patients with early PD were divided into two groups: 110 cases of PD with olfactory dysfunction (PD-OD) and 47 cases of PD without olfactory dysfunction (PD-NOD). The differences in clinical scores and TCS results between the two groups were compared.Results:The midbrain substantia nigra hyperechoic area, substantia nigra hyperechoic positivity rate, third ventricle width, and olfactory dysfunction rate were higher in the PD group compared to the control group, while the midbrain area and olfactory test scores were lower than those in the control group (all P<0.001). The sensitivity and the coincidence rate of TCS combined with the olfactory test for early PD diagnosis (90.0%, 77.1%) were higher than those of TCS alone (60.0%, 71.3%) and olfactory test alone (70.1%, 72.3%), but the specificity (63.7%) was lower than that of both alone (82.8% for TCS and 75.2% for olfactory test), (all P<0.001). MoCA score, visual space and executive ability, memory, attention, and language were positively correlated with the area of the midbrain ( rs=0.38, 0.32, 0.27, 0.25, 0.23; all P<0.05) and negatively correlated with the width of the third ventricle ( rs=-0.39, -0.22, -0.39, -0.22, -0.32; all P<0.05), and orientation was negatively correlated only with the width of the third ventricle ( rs=-0.24, P<0.05). The MoCA score of PD-OD group[22(18, 25)] was lower than that of PD-NOD group[24(20, 26)]( P=0.040). Conclusions:The combination of TCS and olfactory test can enhance the sensitivity and diagnostic agreement rate for early PD diagnosis, providing some auxiliary value. The cognitive function of PD patients is positively correlated with the midbrain area and negatively correlated with the width of the third ventricle. The cognitive function of PD patients with olfactory dysfunction is lower than that of PD patients without olfactory dysfunction. TCS and olfactory test may help assess cognitive function in PD patients.

OBJECTIVETo study the gene mutation in a patient with multiple exostoses, identify the disease-causing gene mutation.

METHODSPolymerase chain reaction and DNA sequencing were used to screen the EXT1 or EXT2 gene mutation, while mismatch primer amplification and restriction endonuclease digestion were performed to confirm the mutation.

RESULTSBy DNA sequencing, a mutation in the seventh intron was detected and located at 26 bp of 3' splice site upstream in EXT1 gene, which was unreported before. Mismatch primer amplification and restriction fragment length polymorphism analysis suggested that this mutation was not detected in the normal control.

CONCLUSIONThe mutation 1633-26(C-->A) may be the disease-causing mutation in this patient with multiple exostoses.

DNA Mutational Analysis ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Mutation ; N-Acetylglucosaminyltransferases ; genetics ; Young Adult

Microtubule is one of the key components of the cytoskeleton and plays an important role in the maintenance of cell shape and the process of signal transduction and mitosis. Due to the extreme importance of microtubule in the process of mitosis, tubulin becomes one of the most important targets for development of new anticancer drugs and tubulin inhibitors are used for the treatment of cancer nowadays. These inhibitors have antitumor activity by inhibiting or promoting the assembly of tubulin to microtubules and interfering the process of cell mitosis. This review summarized the research progress of the tubulin inhibitors, especially the introduction of the tubulin inhibitors of pharmacological activities and the progress of clinical research. Also, the development trend of these inhibitors is discussed.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Microtubules ; drug effects ; metabolism ; Mitosis ; drug effects ; Molecular Structure ; Neoplasms ; drug therapy ; Stilbenes ; chemical synthesis ; chemistry ; pharmacology ; Structure-Activity Relationship ; Tubulin ; metabolism ; Tubulin Modulators ; chemical synthesis ; chemistry ; pharmacology

Objective To explore the factors associated with severe hand-food-mouth disease (HFMD) case in Shanghai.Methods A total of 105 severe HFMD cases diagnosed from May to July,2011 in Shanghai were enrolled as case group while another 210 mild HFMD cases were randomly selected as control group in the same period.All subject' s parents or babysitters were asked to fill in the questionnaire in which including demography,ways of babysitting,behavior and the like.All HFMD cases were diagnosed by both clinical symptom and nuclear acid testing.Data was processed by EpiData (V3.0) and analyzed by SPSS (V17.0).Results Factors as age,gender,Diaspora pattern,migrant,size of house,numbers of family member,numbers of children,frequency of seeing doctor,dishware that sharing with babysitter,food chewed by babysitter,dirty hand,EV71 virus type and diagnosis on HFMD in the fist visit to hospital were found associated with severe HFMD by univariate analysis.Results through multivariate logistic regression showed that factors including:being the only male kid,more than 3 children in the family,dirty hands,unable to be diagnosed as HFMD in the first visit to the hospital,visiting doctor during the past 6 months for 2 and 3 times etc.could be kept in the model with statistical threshold of 0.05.Adjusted ORs and confidence intervals of them were 2.431 ( 1.317-4.487),2.661 (1.332-5.315),3.403 ( 1.871-6.191 ),6.607 (3.011-14.500),2.431 ( 1.111-5.321 ),2.628 ( 1.137-6.071 ) respectively.Being Infected by EV71 was also found a very important risk factor compared with CoxA16 or other enteroviruses,and its adjusted OR was 5.614 (2.409-13.082).Conclusion It was necessary to implement molecular diagnosis for identifying the virus type of HFMD,together with improvement on the capacity of clinical diagnosis in order to diagnose the HFMD cases earlier.More attention should be paid to these HFMD cases with EV71 infection as well as prompting frequent visits to hospitals on those families with more children.

Objective To develop attenuated Salmonella which harboring enterovirus 71 (EV71) VPI gene. Methods The plasmid which expressed VP1 protein of EV71 was constructed by gene recombination. Cellular expression was assessed by Western Blot analysis. The recombinant plasmid was then transformed into attenuated Salmonella SL7207. Results EV71 VP1 gene sequence was inserted into a eukaryotic expression plasmid VR1012. VP1 protein was detected by Western Blot analysis in the culture supernatant. And the attenuated Salmonella harbored the plasmid stable. Conclusion The plasmid was constructed successfully and it can express effectively in vitro. The bacteria which harboring the plasmid were constructed successfully. This has provided a basis for further research of an oral EV71 vaccine.

Objective To develop a system to rescue virus by intracellular expression of T7 RNA Polymerase. Methods The gene of T7 RNA Polymerase was amplified and cloned to VR1012 by molecular biological technology. The expression plasmid VR-1a was then identified. VR-1a and EV71 infectious plasmid were co-transfected in Veto cell. CPE was observed and viral gene viral antigen were detected.Results The gene of T7 RNA Polymerase was successfully cloned into vector VR1012. Vere cell developed to CPE after being transfected VR-1a and EV71 infectious plasmid. EV71 gene was amplified by RT-PCR from the culture. EV71 antigen was also detected by ELISA. Conclusion The method can be used to rescue virus. It could apply to immunologic research of EV71 DNA vaccine.