OBJECTIVETo observe the expressions of caspase-8 and caspase-9 mRNA, and explore the changes of apoptosis of bone marrow hematopoietic cells in patients with chronic mountain sickness (CMS).

METHODSOf 18 CMS patients and 16 controls were enrolled in this study. The apoptotic index (AI) of bone marrow mononuclear cells (BMMNC) was measured by TUNEL technique, the levels of caspase-8 and caspase-9 mRNA in BMMNC of CMS patients and controls were determined by RT-PCR. Results (1)The AI of BMMNC in patients with CMS (8.51 ± 3.35)% was lower than that in controls (16.00 ± 4.28)% (P < 0.01); (2) The values of caspase-8 and caspase-9 mRNA were (0.28 ± 0.07) and (0.23 ± 0.08) respectively, in CMS patients, which were significantly lower than those of (0.45 ± 0.09) and (0.41 ± 0.09) respectively, in the controls (both P < 0.01); (3) Hemoglobin (Hb) value was negatively correlated with levels of caspase-8 and caspase-9 mRNA (r values were -0.52 and -0.61 respectively, both P < 0.05) in CMS patients. There was a negative correlation between AI and Hb (r value was -0.89, P < 0.01) in CMS patients. However, the significant relationship was not found between AI and level of caspase-8 or caspase-9 mRNA (P > 0.05).

CONCLUSIONSThe results showed a decrease apoptosis of BMMNCs and reduced levels of caspase-8 and caspase-9 mRNA in CMS patients, the latter might be involved in the change of BMMNCs apoptosis.

Adult ; Altitude Sickness ; metabolism ; pathology ; Apoptosis ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Humans ; Male ; Middle Aged

OBJECTIVESTo investigate the therapeutic effect of rehabilitation, arthroscopy and "hybrid technique" for posttraumatic knee stiffness (PTKS), and to make the best choice for the treatment.

METHODSFrom February 2004 to November 2009, 66 patients suffered from PTKS were treated, and the clinical data were studied retrospectively, 36 male and 30 female patients with an average age of 41 years were analyzed, knee stiffness time averaged 15 months (0.5 - 108.0 months), 21 cases of patients were treated with rehabilitation (rehabilitation group), 22 cases of patients with arthroscopy + rehabilitation (arthroscopy group) and 23 cases of patients with mini-invasive "hybrid technique" + rehabilitation (hybrid technique group). For each case, the difference of range of motion (ROM) and hospital for special surgery (HSS) score of the knee before and after the treatment were analyzed statistically. The characters of PTKS including the course of the disease, the degree of extensor mechanism involving, physical examination and other ancillary data were also analyzed. The management methods for PTKS were summarized.

RESULTSTotal 66 cases were followed up ranging from 24.0-72.5 months and the mean time was 34.2 months. The average ROM was improved obviously: rehabilitation group increased from 45° ± 22° to 95° ± 24° (t = -11.2, P < 0.05), arthroscopy group from 47° ± 26° to 118° ± 11° (t = -11.0, P < 0.05) and hybrid technique group from 36° ± 22° to 110° ± 14° (t = -13.4, P < 0.05). Both ROM and HSS score of the knee before and after the treatment for each group showed significant difference statistically (t = -9.1, -6.0, -5.2, P < 0.05). Wound necrosis, tearing, re-fracture and extension lag were not found. According to Judet standard at final follow-up, 15 cases were excellent, 3 cases good and 3 cases normal in rehabilitation group; 15 cases were excellent, 5 cases good and 2 cases normal in arthroscopy group; 14 cases were excellent, 8 cases good and 1 case bad.

CONCLUSIONSPathology of PTKS is complex, satisfactory result could be obtained through individualized treatment program, which were established depend on the course of the disease, the degree of extensor mechanism involving, physical examination and ancillary data. The timely and effective surgical interference followed by a comprehensive rehabilitation program is the key point for satisfied outcome.

Adolescent ; Adult ; Aged ; Ankylosis ; etiology ; surgery ; Arthroscopy ; Female ; Follow-Up Studies ; Humans ; Knee Injuries ; complications ; Knee Joint ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Range of Motion, Articular ; Retrospective Studies ; Treatment Outcome ; Young Adult

Objective To evaluate the clinical efficacy and safety of dimemorfan phosphate in the treatment of dry cough.Methods Three hundred seventy -eight patients with dry cough were randomly divided into experimental group ( n=216 ) , positive control ( n=108 ) and place-bo group ( n=54 ).Experimental group was taken dimemorfan phosphate 20 mg, orally, tid, 5 -7 d.Positive control group was taken dextro-methorphan 30 mg, orally, tid, 5 -7 d.Placebo group was taken dimemorfan phosphate simulation tablets 20 mg or dextromethorphan simulation tablets 30 mg, tid, 5-7 d.Comparison of the clinical effica-cy, change of the total cough score after 5-7 d treatment, and the inci-dence of adverse drug reactions between the three groups.Results The clinical effectiveness of cough symptom after treatment , experimental group and positive control group and placebo group were 81.02%, 84.26%and 38.89%, respectively.The difference between experimen-tal group with placebo group was statistically significant ( P <0.01 ) , there was no statistically significant difference between experimental group with positive control group ( P>0.05 ).The average decrease of total cough score after treatment in the experimental group , the positive control group and the placebo group were (2.34 ±1.42), (2.43 ±1.55) and (1.39 ±1.20), respectively.The decrease in the total cough score of experimen-tal group was superior to that of placebo group ( P<0.01 ) , the decrease in the total cough score of experimental group was not inferior to positive control group ( P>0.05 ).The incidence of adverse events were 4.21% in experimental group , 12.04%in positive control group and 5.66%in placebo group , respectively.Among these , the incidence of adverse events associated with drug were 1.87%, 10.19%, 5.66%, respectively.Conclusion Dimemorfan phos-phate has exact antitussive efficacy in the treatment of dry cough , has the same antitussive efficacy and similar side effect in the treatment of dry cough compared with the dextromethorphan.

OBJECTIVETo investigate the clinical features, diagnosis and treatment of glucose transporter 1 deficiency syndrome (GLUT1-DS), as well as the diagnostic value of movement disorders.

METHODSThe clinical data of four children with GLUT1-DS were collected, and their clinical features, treatment, and follow-up results were analyzed.

RESULTSThere were two boys and two girls, with an age of onset of 2-15 months. Clinical manifestations included movement disorders, seizures, and developmental retardation. Seizures were the cause of the first consultation in all cases. The four children all had persistent ataxia, dystonia, and dysarthria; two had persistent tremor, two had paroxysmal limb paralysis, and two had eye movement disorders. Paroxysmal symptoms tended to occur in fatigue state. All four children had reductions in the level of cerebrospinal fluid glucose and its ratio to blood glucose, as well as SLC2A1 gene mutations. The four children were given a ketogenic diet, at a ketogenic ratio of 2:1 to 3:1, and achieved complete remission of paroxysmal symptoms within 5 weeks.

CONCLUSIONSGLUT1-DS should be considered for epileptic children with mental retardation and motor developmental delay complicated by various types of movement disorders. The ketogenic diet is effective at a ketogenic ratio of 2:1 to 3:1 for the treatment of GLUT1-DS.

Carbohydrate Metabolism, Inborn Errors ; diagnosis ; genetics ; therapy ; Child ; Child, Preschool ; Female ; Humans ; Male ; Monosaccharide Transport Proteins ; deficiency ; genetics ; Movement Disorders ; diagnosis ; genetics ; therapy

OBJECTIVETo identify and type three leptospires isolated from Rattus tanezumi in Guizhou Province by using three molecular techniques (PFGE, MLVA, and MLST), reveal the molecular characteristic of causative agents of local leptospirosis and evaluate these three molecular methods based on their detection resolution and efficiency.

METHODSThree Leptospira strains were isolated from the kidney of Rattus tanezumi and cultured with EMJH medium. PFGE, MLVA, and MLST assays were applied to type the three strains isolated from Rattus tanezumi in Guizhou Province.

RESULTSPFGE, MLVA, and MLST typing showed that the three leptospiral isolates matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai. The findings of the genotyping methods were consistent. MLVA and MLST defined genotypes, whereas PFGE allowed the recognition of additional subgroups within the genotypes, and the findings of molecular typing were also consistent with those of traditional techniques.

CONCLUSIONThree leptospiral isolates from Guizhou Province matched with leptospiral serogroup Icterohaemorrhagiae serovar Lai, and PFGE, MLVA, and MLST, as reliable molecular techniques for identifying and typing of Leptospira interrogans, would contribute to the active surveillance, outbreak investigation and source tracking for leptospirosis in Guizhou Province.

Animals ; China ; epidemiology ; DNA, Bacterial ; classification ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Genotype ; Leptospira interrogans ; classification ; genetics ; Leptospirosis ; epidemiology ; microbiology ; veterinary ; Phylogeny ; Rats

OBJECTIVETo determine the impact of Ureaplasma urealyticum (Uu) infection on the integrity of sperm plasma membrane in infertile males.

METHODSSixty-three semen samples were divided into a Uu infection group (n = 32) and a normal control group (n = 31). Conventional semen analyses were performed by computer-assisted semen analysis (CASA) and Uu detected by the culture method. The semen samples were washed with PBS and dyed by SYBR-14/PI double fluorescent staining, followed by detection of the integrity of sperm plasma membrane by flow cytometry. The percentage of the sperm with intact plasma membrane was indicated as the percentage of sperm emitting green fluorescence (SYBR-14+/PI-%).

RESULTSThe Uu infection group showed a significantly decreased integrity of sperm plasma membrane ([45.14 +/- 10.69]%) and reduced percentage of grade a + b sperm ([23.29 +/- 8.81]%) as compared with the normal control group ([72.68 +/- 9.91]% and [46.32 +/- 9.54]%) (P < 0.01). But there were no significant differences in the semen volume, pH value, and sperm concentration between the two groups (P > 0.05).

CONCLUSIONUu infection decreases the integrity of sperm plasma membrane, which might be an important factor of male infertility.

Adult ; Case-Control Studies ; Cell Membrane ; pathology ; Flow Cytometry ; Humans ; Infertility, Male ; microbiology ; pathology ; physiopathology ; Male ; Organic Chemicals ; Semen Analysis ; methods ; Spermatozoa ; metabolism ; pathology ; Ureaplasma Infections ; pathology ; physiopathology ; Ureaplasma urealyticum ; Young Adult

OBJECTIVETo study the clinical effects and application value of intraoperative CT in treatment of severe scoliosis with posterior total pedicle screws.

METHODSThirty-two cases of severe scoliosis were retrospectively analysed in our hospital from June 2009 to June 2011,which were treated by posterior total pedicle screws with intraoperative CT including 12 males and 20 females with an average age of 16.8 years ranging from 10 to 38 years. There were 19 cases combined with thoracic kyphosis among 32 cases. Multiple planar reconstruction technology of intraoperative CT was applied to assess screw position. The numbers (rates) of pedicle screws were calculated and evaluated as different grades in upper thoracic vertebra (T1-T4) ,middle thoracic vertebra (T5-T8), lower thoracic vertebra (T9-T12) and lumbar vertebra. The pedicle screws of 2 grade and 3 grade were defined as malpositioned screws. Times of applicating intraoperative CT were calculated. Cobb angle of all cases and kyphosis angle of the cases combined with thoracic kyphosis were measured before and after surgery. Scoliosis correction rates and kyphosis correction rates were calculated.

RESULTSThere were 686 pedicle screws placed in thoracolumbar of 32 patients (including 544 thoracic pedicle screws,142 lumbar pedicle screws) and 14 patients underwent osteotomy. The rate of malpositioned screws in thoracolumbar was 7.3% by evaluating with intraoperative CT,and it respectively was 5.6%,11.1%, 6.7% and 4.3% in upper thoracic vertebra, middle thoracic vertebra,lower thoracic vertebra and lumbar vertebra. The malpositioned screws were amended in surgery. The mean times of intraoperative CT was 2.6 times (ranged from 2 times to 4 times). The mean preoperative Cobb angle was 95 degrees (ranged from 78 degrees to 123 degrees) and the mean postoperative Cobb angle was 340 (ranged from 19 degrees to 53 degrees). The mean correction rate of Cobb angle was 64%. The mean preoperative kyphosis angle of the patients combined with thoracic kyphosis was 69 degrees (ranged from 46 degrees to 82 degrees) and the mean postoperative kyphosis angle was 32 degrees (ranged from 22 degrees to 45 degrees). The mean correction rate of kyphosis angle was 54%. Four patients suffered cerebrospinal fluid leak after surgery. No infection, vascular lesion and nervous lesion were found. All patients had an average 18-month follow-up (ranged from 12 to 26 months). No broken nails, broken rods and pseudarthrosis were founded.

CONCLUSIONApplication of in traoperative CT in severe scoliosis with posterior total pedicle screws can detect and amend malpositioned screws timely in surgery, to avoid secondary surgery for malpositioned screws and protect the safety of surgery. The effects of surgery is satisfactory.

Adolescent ; Adult ; Bone Screws ; Child ; Female ; Humans ; Male ; Monitoring, Intraoperative ; Retrospective Studies ; Scoliosis ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; methods

OBJECTIVETo investigate the dynamic changes of neurofilaments (NFs) proteins in spinal cords of hens with phenylmethylsulfonyl fluoride (PMSF) pretreatment for exploring the mechanism of tri-o-cresyl phosphate (TOCP)-induced delayed neuropathy (OPIDN).

METHODAdult Roman hens were randomly divided into three groups, control, TOCP and PMSF + TOCP. Birds in PMSF + TOCP set were pretreated with PMSF, 24 hours later, hens in both TOCP group and PMSF + TOCP group were administrated with TOCP at a single dosage of 750 mg/kg. Then all animals were sacrificed on the corresponding time-points of 1, 5, 10, and 21 days respectively after dosing of 750 mg/kg TOCP. The spinal cords were dissected, homogenized, and centrifuged at 100,000 x g. The levels of high molecular neurofilament (NF-H), medium molecular neurofilament (NF-M) and low molecular neurofilament (NF-L) in both pellet and supernatant fractions of spinal cords were determined by SDS-PAGE and Western-blotting.

RESULTSThe hens in TOCP group showed paralysis gait at the end of 21-day experimental period. The levels of NFs proteins in spinal cords changed obviously. Compared with control, the NFs in pellet showed a dramatic decrease on day 10 and then followed by a recovery. In the supernatant, the NFs proteins showed similar changes, which decreased significantly on day 10 and almost recovered control on day 21. Such as, NF-L, NF-M and NF-H decreased by 51%, 86% and 38% on day 10. The OPIDN signs were not observed in PMSF + TOCP group, and imbalances of NFs were obviously alleviated. Compared with control, only NF-M in pellet increased by 21% (P < 0.05) on day 21, others remained no changes; The levels of NF-H and NF-M in supernatant respectively increased by 19% and 35% on day 21, others were no significant statistical differences.

CONCLUSIONTOCP may induce imbalance of NFs levels in progress of OPIDN, and PMSF pretreatment may protect animals from OPIDN by reducing above changes, which may explain that TOCP-induced imbalance of NFs may be connected with the occurrence and development of OPIDN.

Animals ; Chickens ; Female ; Neurofilament Proteins ; drug effects ; Phenylmethylsulfonyl Fluoride ; pharmacology ; Protein Subunits ; drug effects ; Spinal Cord ; drug effects ; metabolism ; pathology ; Tritolyl Phosphates ; toxicity

To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Adjuvants, Immunologic ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology

In order to investigate the influence of cytokine combinations on proliferation and differentiation of human umbilical cord blood CD34(+) cells into megakaryocytes/platelets in vitro, the CD34(+) cells from human umbilical cord blood were amplified in serum-free medium StemSpan(SFEM) supplemented with several cytokine combinations by three-phase culture system. The effects of the cytokine combinations were compared. The results showed that at day 14 of the first culture phase, the CD34(+) cells cultured with cytokine combinations SCF + TPO + FL + IL-3 were amplified (11 000 ± 1 000) times, which were significantly higher than that of cells cultured with SCF + TPO + FL, but were not significantly different from that of cells cultured with SCF + TPO + IL-3 or SCF + TPO + FL + IL-3+ hydroxyl-corticosteroids. At day 7 of the second culture phase, the CD34(+) cells cultured with cytokine combination SCF + TPO + FL + IL-11 were amplified by (204666.7 ± 11718.9) times, which were significantly higher than that of cells cultured with SCF + TPO + FL + IL-3, but were not significantly different from that of cells cultured with SCF + TPO + FL + IL-11 + BMP4 + VEGF. At day 3 and day 6, the CD34(+) platelet-like cells accounted for about (39.8 ± 1.9)%, (39.7 ± 2.6)% and (25.5 ± 1.4)%, (23.1 ± 3.5)% cultured with SCF + TPO + FL + IL-11 and SCF + TPO + FL + IL-11 + BMP4 + VEGF, and significantly higher than that of the cells cultured with SCF + TPO + FL + IL-3. It is concluded that the cytokine combination of SCF + TPO + FL + IL-3 is most suitable cytokines combination for the amplification of CD34(+) hematopoietic progenitor cells. The cytokine combination of SCF + TPO + FL + IL-11 is preferred for the proliferation and differentiation of megakaryocytes, this study lays an experimental basis for investigating the proliferation and differentiation of CD34(+) into megakaryocytes/platelets in vitro.
Antigens, CD34 ; immunology ; Blood Platelets ; cytology ; Cell Differentiation ; Fetal Blood ; cytology ; immunology ; Humans ; Interleukin-11 ; pharmacology ; Interleukin-3 ; pharmacology ; Megakaryocytes ; cytology ; Stem Cell Factor ; pharmacology ; Thrombopoietin ; pharmacology