[Summary] The ossification of the posterior longitudinal ligament ( OPLL ) is caused by environmental , genetic and other factors.With the development of genomics research , researchers have found that mutations are important factors inducing OPLL . Despite the genomics research on OPLL has made important advances , no highly correlation has been found between OPLL and the exact single nucleotide polymorphisms (SNPs) loci.The pathogenesis of OPLL needs further exploration .

OBJECTIVETo investigate different treatment methods for stage-Is testicular mixed germ cell tumors (TMGCTs).

METHODSWe retrospectively analyzed the clinical data about 3'cases of stage-Is TMGCTs (aged 26-39 years) treated in the 175th Hospital of PLA, reviewed relevant literature, and explored the clinical characteristics of TMGCTs.

RESULTSOf the 3 patients, 1 was treated by radical orchiectomy, 1 by radical orchiectomy + retroperitoneal lymph node dissection + BEP chemotherapy scheme, and the other by radical orchiectomy + radiotherapy. The pathological components of TMGCTs were immature teratoma, seminoma, spermatocytoma, chorioepithelioma, embryonal carcinoma, and yolk sac tumor. No recurrence or distant metastasis was found during the 24-month follow-up after surgery.

CONCLUSIONThe diagnosis of TMGCTs primarily depends on physical examination, ultrasonography, MRI, and measurement of serum tumor markers, while its confirmation necessitates pathological examination, and its treatment is basically radical orchiectomy.

Adult ; Carcinoma, Embryonal ; pathology ; Endodermal Sinus Tumor ; pathology ; Humans ; Lymph Node Excision ; Male ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Neoplasms, Germ Cell and Embryonal ; pathology ; surgery ; Orchiectomy ; Retrospective Studies ; Seminoma ; pathology ; Teratoma ; pathology ; Testicular Neoplasms ; pathology ; surgery

Objective:To investigate the regulation effect of biological intensity electric field (EF) on the motility and CD9 expression of human epidermal cell line HaCaT and mouse epidermal cells.Methods:The experimental research method was used. Human epidermal cell line HaCaT cells in logarithmic growth phase and primary mouse epidermal cells isolated from 16 BALB/c mice aged 1-3 days were used for the experiment. HaCaT cells were divided into EF group treated with EF in the intensity of 200 mV/mm and sham EF group treated with simulated operation. The cell migration (displacement velocity, trajectory velocity, and direction, with 46 samples in EF group and 34 samples in sham EF group) and arrangement were observed in the living cell workstation, and the distribution and expression of CD9 protein were detected by immunofluorescence method. Both HaCaT cells and mouse epidermal cells were divided into sham EF group (simulated operation) and 50 mV/mm group, 100 mV/mm group, 200 mV/mm group and 400 mV/mm group treated with EF in the corresponding intensity respectively. Both HaCaT cells and mouse epidermal cells were divided into blank control group without any treatment and 1 h group, 3 h group and 6 h group treated with EF in the intensity of 200 mV/mm for corresponding time respectively. The expression of CD9 protein was detected by Western blotting (n=3). Data were statistically analyzed with Mann-Whitney U test, one-way analysis of variance, t test and least significant difference test. Results:Within 3 hours of treatment, HaCaT cells in EF group tended to move towards the negative electrode obviously, while HaCaT cells in sham EF group moved randomly around the origin; compared with those of sham EF group, the directivity of HaCaT cells in EF group was significantly enhanced, and the displacement velocity and trajectory velocity were significantly increased (Z=-3.975, -6.052, -6.299, P<0.01). After 3 hours of treatment, the long axis of HaCaT cells in EF group was perpendicular to the direction of EF, while HaCaT cells in sham EF group arranged randomly. After 3 hours of treatment, the expression of CD9 protein in HaCaT cells in EF group was significantly down-regulated compared with sham EF group (t=4.527, P<0.01), although both expressed on cytomembrane. After 3 hours of treatment, the expression of CD9 protein in HaCaT cells and mouse epidermal cells in sham EF group, 50 mV/mm group, 100 mV/mm group, 200 mV/mm group and 400 mV/mm group were 0.332±0.021, 0.283±0.032, 0.254±0.020, 0.231±0.041, 0.212±0.031 and 0.565±0.021, 0.453±0.022, 0.389±0.020, 0.338±0.021, 0.233±0.011, respectively. For both types of cells, compared with that of sham EF group, the expression of CD9 protein in cells was significantly decreased in the four groups of EF treatment (P<0.01); compared with that of 50 mV/mm group, the expression of CD9 protein in cells was significantly decreased in the other three groups of EF treatment (P<0.01); compared with that of 100 mV/mm group, the expression of CD9 protein in cells was significantly decreased in 200 mV/mm group and 400 mV/mm group (P<0.01); compared with that of 200 mV/mm group, the expression of CD9 protein in cells was significantly decreased in 400 mV/mm group (P<0.01). The expression levels of CD9 protein in HaCaT cells and mouse epidermal cells in blank control group, 1 h group, 3 h group and 6 h group were 0.962±0.031, 0.784±0.020, 0.531±0.021, 0.409±0.011 and 0.963±0.031, 0.872±0.031, 0.778±0.040, 0.591±0.041, respectively. For both types of cells, compared with that of blank control group, the expression of CD9 protein in cells was significantly decreased in 1 h group, 3 h group, and 6 h group (P<0.01); compared with that of 1 h group, the expression of CD9 protein in cells was significantly decreased in 3 h group and 6 h group (P<0.05 or P<0.01); compared with that of 3 h group, the expression of CD9 protein in cells was significantly decreased in 6 h group (P<0.01).Conclusions:The biological intensity EF can induce the directional migration and arrangement of HaCaT cells and down regulate the expression of CD9 in HaCaT cells and mouse epidermal cells in a time-dependent and intensity-dependent manner.

OBJECTIVETo study the characteristics of thyroglossal tract cyst epidemiology and summarize how to improve the operation therapeutic effect.

METHODSThe clinic appearance, the pathological characteristics and the therapeutic effect of the 60 thyroglossal tract cyst cases were analyzed.

RESULTSThyroglossal tract cyst mostly occurred in children and teenagers. The incidence of female patients was more than that of male patients. Most of thyroglossal tract cyst located in the middle line of the neck and were explored tubular-objects and fibroropes. The histopathological feature was variform.

CONCLUSIONThe clinical appearance and pathology characteristic of thyroglossal tract cyst are variform. Thyroglossal tract cyst may be recurred easily if it is not operated well.

Adolescent ; Child ; Female ; Humans ; Male ; Thyroglossal Cyst

The aim of the research is to investigate the genetic characteristics of Legionella pneumophila serogroup1 (LP1)in Sichuan Province.The sequence-based typing (SBT) and multiple-locus VNTR analysis (MLVA) were used to describe the genetic polymorphism of 42 strains which were isolated from 1989-2016 in Sichuan Province,China.According to the reference,PCR was used to detect the 8-VNTR loci and 7 housekeeping genes respectively.The VNTR results were determined by using capillary electrophoresis,and the SBT results were sequenced and compared with the database of EWGLI.Results showed that totally 42 stains were divided into 8 MLVA types with the advantage types were M08 (47.6 ％) and M07 (23.8 ％).Twelve ST types were obtained with 3 main clonal complex and 2 singleton,including 2 novel ST types,among those,ST1 was the predominant type,accounting for 52.3 ％,following by ST630 (14.2 ％).In conclusion,our results demonstrated MLVA and SBT were both applied to the research for molecular epidemiological investigation of LP1 and showed the high genetic polymorphism and the regional specificity.The results also suggest that the isolates are a potential threat to the public,effective control and prevention strategies are urgently needed.

Objective To explore the rule of transportation in extracellular space (ECS) and the changes induced by the external stimulation with tracer-based MRI.Methods Thirty two mature Sprague Dawley rats were randomly divided into four groups,i.e.caudate nucleus-control (Cc) group,thalamus-control (Tc) group,caudate nucleus-moving (Cm) group and thalamus pain stimulation (Tp) group.The rats were anesthetized and a series of MR scanning were performed before and after the injection of Gd-DTPA in ECS of caudate nucleus and thalamus until the intensity of Gd-DTPA was invisible.Half-life of Gd-DTPA in ECS was calculated and analyzed with t-test.Results Gd DTPA in caudate nucleus was transported to the ipsilateral cortices away from the injection points,which only distributed on site in the thalamus.The transportation between the two partitions was not observed.The half-life of Tc group ([49.93±2.11] min) was significantly shorter than that of Cc group ([104.30±54.12];t=2.839,P＜0.05),no difference was observed between the Cm group ([113.42±47.32]min) and Cc group (t=0.359,P＞0.05),while half-life of Tp group ([109.40±10.33]min) was significantly longer than that of Tc group (t=15.954,P＜0.05).Conclusion Tracer-based MRI can be used to investigate the rule of transportation in ECS,and the transportation and clearance of substances into the ECS can be influenced by a selective external stimulation.

OBJECTIVETo explore the characteristics of severe acute respiratory syndrome (SARS) transmission in the population base on analyzing the first imported case and the chains of transmission.

METHODSFor the first imported SARS case and cases who were transmitted by the index case, epidemiological investigations were conducted using the guidelines for surveillance and case investigation issued by the Ministry of Health. Data as the date of onset of symptoms, date of hospitalization, contact history etc. for each of the cases and their close contacts were collected and analyzed.

RESULTSThe first imported SARS case introduced to Beijing had infected 9 people within the family and at the hospital, with two of whom died of the same disease. The incubation period for that index case was 4 days, and that for the cases considered to be the secondary and tertiary generations were 7 and 8 days, respectively. The shorter the incubation period, the longer the fever would last and clinically more severe.

CONCLUSIONOne of the epidemiological characteristics of SARS in Beijing was noticed that the disease clustered in families and hospitals. Infection through droplets and close contact has been viewed as the primary mode of transmission.

Adult ; China ; epidemiology ; Contact Tracing ; Cross Infection ; transmission ; Family Health ; Female ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission

The recombinant plasmid carrying the gene encoding 3C protease of Enterovirus 71 (EV71) was constructed, the recombinant protein was then expressed and purified, the functional activity was also measured. Firstly, the 3C protease gene was inserted into pET28a vector, the constructed recombinant plasmid was transformed into E. coli BL21 (DE3) for expression under the induction of IPTG. The expressed protein was purified by affinity chromatography (Ni-NTA) and the N-terminus His-tag was cleaved by enterokinase from 3C protease. The activity of 3C protease was evaluated with fluorescent peptide substrates. It was verified by restriction analysis and sequencing that recombinant plasmid pET28a-3C was constructed correctly and functionally expressed in E. coli BL21 (DE3) resulting in the production of recombinant 3C protease with a size of 22kD. Both His-tag and non-His-tag (cleaved by enterokinase) 3C protease exhibited similar enzyme activity to 3B-3C fluorescent peptide with Km, Vmax and Kcat values of 22 microM, 434nM. Min(-1) and 0.0669 Min(-1), respectively. The optimial pH and temperature were 7.0 and 30-37 degrees C, respectively. The acquirement of recombinant purified 3C protease with high activity has paved the way of further studies on anti-viral inhibitors, structural protein assembly, vaccine development and detection methods of EV71.
Cloning, Molecular ; Cysteine Endopeptidases ; chemistry ; genetics ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Kinetics ; Viral Proteins ; chemistry ; genetics ; metabolism

OBJECTIVETo discuss the biological function and regulation mechanism of curcumin in promoting human colorectal carcinoma (LoVo) cells apoptosis.

METHODConventional in vitro culture in human colorectal carcinoma cells LoVo, When 80%-90% confluence was reached, cells were treated with curcumin at different concentrations (0-20 mg x L(-1)). Curcumin's effect on cell proliferation level was examined by MTT colorimetry. The ultrastructure of curcumin-treated LoVo cells were observed with transmission electron microscope (TEM). The amount of PI-positive LoVo cells after the curcumin treatment were determined by flow cytometry. The cell apoptosis rate was detected by Annexin V-FITC/PI double staining. The mRNA level of Bax, Bcl-2, Caspase-3 and Bcl-xL were tested by means of RT-PCR.

RESULTMTT test indicates curcumin could inhibite the growth and proliferation of LoVo cells in a time- and concentration-dependent manner. TEM examination showed that curcumin can make LoVo cell morphological changes, showing the typical characteristics of apoptotic cells. Flow cytometry instrument analysis showed that curcumin can arrest cell cycle at S phase, and induce apoptosis of LoVo cells. RT-PCR test showed that curcumin can activate the expression of Bax and Caspase-3, inhibit the expression of Bcl-2 and Bcl-xL at the mRNA level.

CONCLUSIONCurcumin can significantly inhibit the proliferation and induce the apoptosis of human colorectal carcinoma cells LoVo. Such biological effect may be associated with activating Caspase-3 signal channel by activating Bax expression and inhibiting Bcl-2 and Bcl-xL expression. This study lays an important foundation for further discussing the mechanism of curcumin in inducing human colorectal carcinoma LoVo apoptosis.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; drug therapy ; pathology ; Curcumin ; pharmacology ; Dose-Response Relationship, Drug ; Humans

OBJECTIVETo establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.

METHODSPrimers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.

RESULTSThe 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.

CONCLUSIONThe established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.

Astroviridae ; genetics ; isolation & purification ; Diarrhea ; virology ; Feces ; virology ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Norovirus ; genetics ; isolation & purification ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Sapovirus ; genetics ; isolation & purification