Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.

Objective To explore the clinic application value of ultrasonography examination of optic nerve sheath diameter(ONSD) in brain injury.Methods From July 2008-June 2011,90 cases of brain injured patients were chosen as experimental group including light (A group),medium (B group),and heavy (C group) brain injured patients according to the admission GCS score ;50 cases of conventional physical examination and 90 cases of volunteers 50 in neurosurgical outpatient were chosen as control group.The ONSD of both groups were measured 3 mm behind the globe through orbital using color sonographic with different time after admission.3 times measurements were carried out for every optic nerve sheath.All client's ONSD mean and standard deviation were calculated.In 0.5 h after color dopplar ultrasound examination,lumbar vertebra puncturing measured intracranial pressure in different groups.Results After admission (1d,3 d,7 d,14 d),the ONSD of A group was (4.54 ±0.32)mm,(4.42 ±0.30)mm,(4.44 ±0.32) m,and (4.43 ± 0.25) mm,respectively; The ONSD of B groups was (4.48 ± 0.28) mm,(4.52 ± 0.24) mm,(4.46 ±0.28)mm,and (4.38 ±0.22)mm,respectively; The ONSD of C group was (5.67 ±0.35)mm,(6.36 ± 0.42) mm,(5.65 ± 0.23) mm,and (4.76 ± 0.35) mm,respectively.After admission (1 d,3 d,7 d,14 d),the intracranial pressure (IP) of A group was (82 ± 11) mmH2O,(79 ± 12) mmH2O,(90 ±15) mmH2O,and (86 ± 14) mmH2O,respectively; The IP of B group was (78 ± 15) mmH2O,(85 ± 10)mmH2O,(78 ± 16) mmH2O,(80 ± 11) mmH2O,The IP of C group was (225 ± 26) mmH2 O,(288 ± 23)mmH2O,(256 ± 23) mmH2O,(122 ± 18) mmH2O,respectively.Group D had the ONSD average of (4.58± 0.41)mm and IP of (88 ± 10)mmH2O after eyeball 3-mm place.No difference was found between A and B,A and D,or B and D (P＞0.05) ; A difference was found between A and C,B and C,or D and C (t =12.24～24.67,P＜0.01).Conclusions The ONSD and IP in light medium brain injured patients had no change.In patients with severe brain injury,IP changed with the time after injury,the ONSD increased with the IP,the ultrasonography examination of ONSD with the important value in the diagnosis and treatment can respond the IP increase,which is a non-invasion,convenient,fast,and feasible method for evaluation of cranial high pressure.

Objective To investigate the factors associated with the occurrence of pulmonary complication in patients with cervical spine fiactures and concurrent cervical cord injury in Wenchuan earthquake. Methods A retrospective analysis was conducted among the 9 patients with cervical spine fractures and cervical cord injuries treated in our department between May 12 to August 6, 2008. Results All the patients received surgical treatment for cervical spine fractures and cervical cord injuries. Six of the patients developed pulmonary complications 5 days after the injury, including 3 patients with pneumonia, 2 with ventilation disorder, and 1 with lung edema and hemopnenmothorax. Aggressive respiratory management was administered in these patients, and the pulmonary complications were effectively controlled. Conclusion Patients with cervical spine fractures and concurrent cervical cord injury often experience severe pulmonary complications during the acute phase (<5 days), which can be more likely in patients with high level injury, chest trauma, old age, preexisting pulmonary illnesses or smoking history. Early detection of the complications results in better therapeutic effect with conservative therapy.

To investigate the purging effect of CD3AK/iNOS on primary leukemic cells from chronic myeloid leukemia patients in vitro, amphotropic packaging cell line PA317 transfected with the whole length of iNOS gene was cultivated, amplified and screened by G418. The viral titer was determined by the NIH3T3 cells. Human peripheral blood mononuclear cells were isolated and activated by anti-CD3 monoclonal antibody in vitro. CD3AK cells were incubated with viral supernatant and selected by G418. Resistant clones were assayed for iNOS gene expression by RT-RCR. The content of nitric oxide and the activity of iNOS in the culture supernatant of CD3AK/iNOS were evaluated by the method of Griess. After BMMNC or PBMNC from CML patients were co-cultured with CD3AK/iNOS, CD3AK/Neo and CD3AK/iNOS respectively, the expression of bcr/abl fusion gene was detected by serial dilution semi-quantitative net RT-PCR assay. The results showed that anti-G418 positive packaging cell line PA317 transfected with the whole length of iNOS gene clones could stably synthesize and excrete recombinant retroviral vectors. The titer of recombinant retroviral vectors was 1.0 x 10(5) CFU/ml. After being transfected by recombinant retroviral supernatant, the iNOS cDNA was expressed in CD3AK/iNOS. The content of NO and activity of iNOS that synthesized and excreted by CD3AK/iNOS were notably increased, compared with those of CD3AK. There were statistically significant differences in NO content and iNOS activity between two groups. After BMMNC or PBMNC from CML patients were co-cultured with CD3AK/iNOS, CD3AK/Neo and CD3AK/iNOS respectively, the expression of bcr/abl fusion gene in all of them was down-regulated by serial dilution semi-quantitative RT-PCR assay. It is concluded that construction of CD3AK/iNOS can markedly increase the content of NO and the activity of iNOS, which can be more efficient in in vitro purging leukemia cells for autologous hematopoietic stem cell transplantation.
Adult ; Aged ; Animals ; CD3 Complex ; immunology ; Cell Line ; Cytotoxicity, Immunologic ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; pathology ; Male ; Mice ; Middle Aged ; NIH 3T3 Cells ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured

Objective?To investigate the feasibility, safety, operating essentials and the short-term therapeutic effect of total laparoscopic resection for colorectal cancer by Natural Orifice Specimen Extraction (NOSES).?Methods?The clinical data of 50 patients underwent total laparoscopic resection for colorectal cancer by NOSES from January 2016 to June 2017 were retrospectively analyzed.?Results?All of the 50 patients with colorectal cancer successfully received total laparoscopic resection by NOSES. None of the patients had serious postoperative complications and death related to the operation. The average operating time was (140.0 ± 29.0) minutes. The blood loss was (70.0 ± 23.4) ml. The number of lymph nodes harvested were (14.0 ± 2.3). There was no bacteria infection in abdominopelvic cavity post-operation and no recurrence occurred until the end of postoperative follow- up. Postoperative time of bed rest was (2.0 ± 0.5) days. The time of the first anal exhaust was (2.0 ± 0.5) days. The postoperative hospitalization stay was (8.5 ± 3.0) days. There was none case of lung infection while one case of anastomotic leakage. No cancer cells remained in resection margin. No local recurrence and metastasis was found in all patients after follow-up for 3 to 24 months.?Conclusion?Total laparoscopic resection for colorectal cancer by NOSES is safe and feasible, and has the advantage of minimally invasive, less pain, rapid rehabilitation, good cosmetic effect and less postoperative complications.

To study the effects of sodium valproate (VPA) on human myelodysplastic syndrome cell line MUTZ-1. The cell proliferation was determined by MTT assay, apoptotic morphological features were observed by light microscopy and transmission electronmicroscopy, cell apoptosis and cell cycle shift were analyzed by flow cytometry (FCM). The results showed that VPA could inhibit the growth of MUTZ-1 cells in dose-and time-dependent manners. The typical apoptotic morphological features appeared in MUTZ-1 cells treated with 4 mmol/L VPA for 72 hours. Pyknosis of cells and nuclei, disintegration of nuclear chromatin and apoptotic body could be observed by light microscopy. Aggregation and margination of nuclear chromatin, concentration of plasm, increment of density and chromatin mass of irregular size could be observed by transmission electronmicroscope. The flow cytometric analysis indicated that the VPA could induce cell apoptosis, apoptosis rate increased in dose-dependent manner, ratio of cells at G(0)/G(1) phase increased and ratio of cells at S phase decreased in dose-dependent manner, the cells were arrested at G(0)/G(1) phase. It is concluded that the VPA can induce apotosis and inhibite proliferation of MUTZ-1 cells via arresting cells at G(0)/G(1) phase.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line ; Dose-Response Relationship, Drug ; Humans ; Myelodysplastic Syndromes ; pathology ; Valproic Acid ; pharmacology

Animals ; Cells, Cultured ; Chitosan ; pharmacology ; DNA, Viral ; isolation & purification ; Hepatocytes ; virology ; Nanofibers ; Primary Cell Culture ; Swine ; Transplants ; virology

This study was aimed to establish a cytokine-independent human myelodysplastic cells line from bone marrow of a patient with MDS-CMML. This cell line was incubated in mixed culture of RPMI 1640 and DMEM with 15% bovine serum, but without cytokines; its biological characteristics were identified by morphology, surface marker profiles, cell proliferation, differentiation and apoptosis. The results showed that the established cell line could not depend on cytokines for long-term survival and growth, and could differentiate into colony-forming unit-macrophage, colony-forming unit-megakaryocyte. In conclusion, a cytokine-independent human myelodysplastic syndrome cell line, named MDS-JSN04 (MDS Nanjing Jiansu 04), was established. Its partial biological characteristics were identified and clarified.
Antigens, CD19 ; analysis ; Antigens, CD20 ; analysis ; Apoptosis ; drug effects ; Bone Marrow Cells ; metabolism ; pathology ; ultrastructure ; CD79 Antigens ; analysis ; Cell Differentiation ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Culture Media ; pharmacology ; Cytokines ; pharmacology ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Humans ; Lewis X Antigen ; analysis ; Male ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Sialic Acid Binding Ig-like Lectin 2 ; analysis ; Time Factors

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

BACKGROUNDValproic acid (VPA) improves early survival and organ function in a highly lethal poly-trauma and hemorrhagic shock model or other severe insults. We assessed whether VPA could improve organ function in a rat model of septic shock and illustrated the possible mechanisms.

METHODSForty Sprague-Dawley rats were randomly assigned to four groups (n = 10): control group, VPA group, LPS group, and LPS + VPA group. Lipopolysaccharide (LPS) (10 mg/kg) was injected intravenously to replicate the experimental model of septic shock. Rats were treated with VPA (300 mg/kg, i.v.) or saline. Six hours after LPS injection, blood was sampled for gas analysis, measurement of serum alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine and tumor necrosis factor-alpha. Lung, liver and kidney were collected for histopathological assessment. In addition, myeloperoxidase activity and tumor necrosis factor-a in pulmonary tissue were measured. Acetylation of histone H3 in lung was also evaluated by Western blotting.

RESULTSLPS resulted in a significant decrease in PaO2, which was increased by VPA administration followed LPS injection. In addition, LPS also induced an increase in the serum levels of alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine, and tumor necrosis factor-alpha. However, these increases were attenuated in the LPS + VPA group. The lungs, liver and kidneys from the LPS group were significantly damaged compared with the control group. However, the damage was attenuated in the LPS + VPA group. Myeloperoxidase activity and tumor necrosis factor-alpha levels in pulmonary tissue increased significantly in the LPS group compared with the control group. These increases were significantly inhibited in the LPS + VPA group. Acetylation of histone H3 in lung tissue in the LPS group was inhibited compared with the control. However, the level of acetylation of histone H3 in the LPS + VPA group was markedly elevated in contrast to the LPS group.

CONCLUSIONSTreatment with VPA can attenuate multiple organ damage caused by LPS induced septic shock. Our data also suggest that the beneficial effects are in part due to the decrease in inflammatory cytokines and restoration of normal acetylation homeostasis.

Acute Kidney Injury ; drug therapy ; metabolism ; Animals ; Blotting, Western ; Chemical and Drug Induced Liver Injury ; drug therapy ; metabolism ; Lung Injury ; drug therapy ; metabolism ; Male ; Multiple Organ Failure ; drug therapy ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; drug therapy ; metabolism ; Valproic Acid ; therapeutic use