Objective To observe the dynamic serological test results of 28 patients with acute brucellosis,and to investigate the relationship between serological test results and diagnosis,curative effect and prognosis of brucellosis.Methods Twenty eight patients(infected with sheep brucellosis) with acute brucellosis in the Department of Brucellosis in Heilongjiang Provincial Land Reclamation Headquarters General Hospital were selected as research subjects,and their serological changes were tested by means of tube agglutination test (SAT)and clinical outcomes were compared before and after each stage of treatment.In addition,symptoms of fever,weakness,sweating,joint pain,swollen lymph nodes and biochemical parameters [alanine aminotransferase (ALT),lactate dehydrogenase (LDH)] were also tested 3,6 and 9 weeks after the treatment.Results Antibody titer reached the peak at the third week,1 ∶ 400(++),which accounted for 39.29％ (11/28); 2857％(8/28) of the patients became negative at the sixth week; 50.00％ (14/28) became negative at the ninth week.Before the treatment,20(71.43％) patients had the symptom of fever,8 (28.57％) patients had the symptom of hyperhidrosis,28 (100.00％) patients had the symptom of joint pain,7(25.00％) patients had the symptom of lymph node enlargement,28 (100.00％) patients' ALT was elevated,and 26(92.86％) patients' LDH was elevated.After three weeks of treatment,except the three patients (10.71％) who occasionally had fever,the rest of the patient's temperature was returned to normal.Also the numbers of patients with the symptoms of fatigue,sweating and joint pain were significantly reduced,and specifically,the conesponding number was 13(46.43％),2(7.14％),and 21 (75.00％)patients,respectively.ALT and LDH returned to normal(only one patient's ALT was out of the range).At the sixth week,all the patients' symptoms of fever and hyperhidrosis disappeared.The number of patients with the symptoms of joint pain and lymph node enlargement reduced to 12(42.86％) and 3(10.71％),respectively.The results of biochemical tests(ALT and LDT) returned to normal.At the ninth week,most patients' clinical symptoms disappeared.A few patients still had the symptoms of weakness[2(7.14％)] and joint piin[6(21.43％)].Conclusions After effective treatment,antibody titer of patients decreases rapidly,at the same time,the clinical symptoms improve quickly.There is a parallel relationship between the change of antibody titer and clinical symptoms.It is demonstrated that the appearing time of brucellosis specific antibodies,the ampfitude and speed of change of antibody titers can be used in diagnosis,therapeutic evaluation and prognosis of the disease.

Objective To observe the clinical features of brucellosis spondylitis and analyse the reasons for its misdiagnosis,and improve the level of diagnosis and differential diagnosis.Methods Forty-two clinically diagnosed patients with brucellosis spondylitis were studied retrospectively,and these patients were diagnosed and hospitalized in the General Hospital of Heilongjiang Land Reclamation Bureau.Their medical records were analyzed,which included the general information,medical history,clinical symptoms,results of magnetic resonance imaging(MRI) and serum tube agglutination test(SAT) and so on.Results Main clinical symptoms and signs were severe persistent neck,back and leg pain.They also had plate shape low back but without kyphosis.In addition,patients had to keep in one posture because their spinal activity was limited.Also,scoliosis or pelvic tilt and lameness may occur when standing,which were typical symptoms of nerve root compression.Thirteen cases were diagnosed as tuberculosis,accounting for 30.95％(13/42); 6 cases were diagnosed as lumbar disc herniation,accounting for 14.28％ (6/42); 2 cases were diagnosed as ankylosing spondylitis,accounting for 4.76％ (2/42).Therefore,the total rate of misdiagnosis was 50％ (21/42).Abnormal MRI signal intensity can be seen in the pathological vertebrae.Specifically,T1-weighted images (T1WI) showed low signal,T2-weighted images (T2WI) showed high signal,or mixed high and low signal intensity was observed.Vertebral showed wedge deformation without collapse and sequestrum; strip and sheet abnormal signal can also be found within the intervertebral disc.Normal structure disappeared and disc space became narrow.Accordingly,the plane dural sac was compressed to form visible abscess near the spine,but psoas abscess was not found.Patients with positive SAT result accounted for 92.85％ (39/42).Conclusion Reasons for misdiagnosis include lack of detailed medical records,atypical clinical symptoms and similar imaging changes and so on.

Objective To discuss a real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) wether if can be used to detect Brucella. Methods According to the BCSP31 gene sequences specific for Brucella, one pair of primers and one TaqMan probe were designed. A real-time PCR was developed with the BCSP31 fragments cloned into PMD18-T vector. The standard cure was established and the sensitivity, the species specificity and the stability of the assay were evaluated. The clinical blood specimens were detected by QT-PCR and compared with clinical diagnosis. Results The standard curve was established with the standard template and the relationship between the Ct and the DNA copy number was linear(r=0.999). The sensitivity of the real-time PCR was 5 copies/μl. The sensitivity of the common PCR was 5×102 copies/μl. The sensitivity was about 100 times higher than common PCR. Species specificity of this FQ-PCR assay evaluated using genomic DNA from 6 Bmcella strains and 5 non-Brucella strains and strong fluorescence was detected in all Brucella strains. The CV of intra-assay and inter-assay reproducibility were 0.71%,7.23%, reprectively. Twenty-four specimens from clinical brucellosis cases, 19 showed positive, the positive coincident rate was 79%(19/24). The negative results were obtained for all 31 negative control, and the negative coincident rate was 100%(31/31). Two were positive from all 30 specimens clinically suspected. Conclusions Highly specific, sensitive, repeatable and coincidental with clinic, this FQ-PCR is quite useful for rapid detection of tiny DNA of Brucella in various samples and laboratory diagnosis.

Objective To develop a loop-mediated isothermal amplification (LAMP) method for rapid diagnosing of Legionella pneumophila in the Pathogen Detection Department(PDD) or in small-scale laboratory. Methods Five primers (2 Inner Primers, 2 Outer Primers and a Loop Primer) for the LAMP test were designed by targeting the mip gene of Lpneumophila and reaction system of LAMP reaction was optimized. 12 strains of L.pneumophila, 45 local strains, 6 non-L.pneumophila strains, 11 other strains and 59 environmental water samples were analyzed to evaluate the specificity and sensibility of the LAMP amplification. At the same time, the results of the LAMP were also compared with biochemical culture and quantitative PCR methods. Results The amplification products of L.pneumophila turned green by visual inspection and had ladder-like pattern on the gel, but non-L.pneumophila and other products from the strains remained orange by visual examination and had no band on the gel. The detection rate of LAMP was higher than the biochemical culture and the real-time PCR methods. Reaction time of the LAMP method was only 1.5 h and the detection limit of LAMP assay was 5 cfu/reaction. In addition, the LAMP results could be determined only by visual inspection. Conclusion LAMP assay targeting the mip gene of L.pneumophila appeared to be rapid, specific, and sensitive for the detection of L.pneumophila. This method not only reduced the dependence of complicated equipment but also had a potential for wider use in the PDD, small-scale laboratory, emergency motor vehicle or for field survey.

OBJECTIVETo evaluate the influence of glucose-insulin-potassium treatment (GIK) on the levels of inflammatory cytokines in the scalded rats with MODS.

METHODSOne hundred and twenty Sprague Dawley rats were inflicted with 30% TBSA full-thickness scalding, and MODS model was reproduced with intraperitoneal injection of endotoxin following burn injury. Then the rats were randomly divided into GIK, glucose (G) and control (C) groups, with 40 rats in each group. The serum contents of glucose, lactate acid, TNF-alpha, NO and IL-6 of the rats in the three groups were determined during 1 to 7 PSD, and the mortality rate within 7 PSD was observed.

RESULTSThe serum contents of glucose, lactate acid, TNF-alpha, NO and IL-6 of the rats in the GIK group were obviously lower than those in the other two groups during 1 to 7 PSD (P < 0.01), and reached the lowest level at 6 to 7 PSD (TNF-alpha: 2.37 +/- 0.54 microg/L; IL-6: 0.28 +/- 0.17 microg/L; NO: 29 +/- 9 micromol/L). The content of glucose and lactate acid in G group were obviously higher than those in control group (P < 0.01), but the contents of TNF-alpha, IL-6 and NO content were similar between these two groups (P > 0.05). The mortality in GIK group within 7 PSD was 20.0%, which was evidently lower than that in G (37.5%) and C (47.5%) groups (P < 0.05), while that between G and C groups was similar (P > 0.05).

CONCLUSIONThe administration of GIK might ameliorate sepsis by reducing the levels of inflammatory cytokine after burns and endotoxin challenge.

Animals ; Blood Glucose ; metabolism ; Burns ; complications ; diagnosis ; metabolism ; Cytokines ; metabolism ; Disease Models, Animal ; Glucose ; therapeutic use ; Insulin ; therapeutic use ; Lactic Acid ; blood ; Multiple Organ Failure ; diagnosis ; etiology ; metabolism ; Potassium ; therapeutic use ; Prognosis ; Rats ; Rats, Sprague-Dawley

Bruceae Fructus is a Chinese herbal medicine with the chemical constituents mainly including Brucea javanica oil, quassinoids, flavonoids, steroids, triterpenoids, and alkaloids. Modern research demonstrates that Bruceae Fructus has anti-tumor, anti-malaria, anti-inflammatory, and lipid-lowering activities. This paper introduces the resource distribution, chemical constituents, and pharmacological activities of Bruceae Fructus. Further, according to the concept of quality marker(Q-marker), this paper predicts the Q-markers of Bruceae Fructus from the specificity of chemical components, pharmaceutical activity, measurability of chemical constituents, compatibility, and clinical efficacy, aiming to provide a theoretical basis for establishing the quality standard of Bruceae Fructus.
Fruit ; Drugs, Chinese Herbal/pharmacology* ; Quassins ; Flavonoids ; Biomarkers

Objective: To investigate the influence of the degrees of intravesical prostatic protrusion (IPP) on the recovery of urinary continence after radical prostatectomy. METHODS: We retrospectively analyzed the clinical data on 212 patients diagnosed with prostate cancer by biopsy and treated by laparoscopic radical prostatectomy by the same surgeon. Based on the degrees of IPP measured by MRI, we divided the patients into an IPP ≤ 10 mm group (n = 146) and an IPP > 10 mm group (n = 66) and determined the factors influencing the recovery of urinary continence by univariate and multivariate logistic regression analyses. RESULTS: At 1, 3, 6 and 12 months after surgery, the urinary continence rates of the patients were 32.5%, 50.5%, 82.1% and 91%, respectively. Univariate analysis indicated that the factors influencing the recovery of urinary continence included IPP, body mass index (BMI), bladder neck preservation (BNP), neurovascular bundle preservation (NVBP) and clinical tumor (T) stage at 3 months (P < 0.05 or P < 0.01), age, IPP, BMI, BNP and clinical T stage at 6 months (P < 0.05 or P < 0.01), and age, IPP, BMI, BNP, NVBP and clinical T stage at 12 months (P < 0.05), while multivariate logistic regression analysis showed the independent influencing factors to be IPP > 10 mm (P < 0.001), BMI ≥ 25 kg/m2 (P = 0.004) and BNP (P = 0.032) at 3 months, and IPP and BMI at 6 months (both P < 0.01) and 12 months (P < 0.01 and P = 0.033). CONCLUSIONS IPP > 10 mm and BMI ≥ 25 kg/m2 are independent factors influencing the long-term recovery of urinary continence after radical prostatectomy.

The main pathogenesis of saphenous vein graft neointimal hyperplasia after coronary artery bypass grafting (CABG) is inflammation-caused migration and proliferation of vascular smooth muscle cells (VSMCs). Janus kinase 2/signal transducer and activators of transcription 3 (JAK2/STAT3) pathway is an important signaling pathway through which VSMCs phenotype conversion occurs. Suppressor of cytokine signaling 3 (SOCS3) is the classic negative feedback inhibitor of JAK2/STAT3 pathway. Growing studies show that SOCS3 plays an important anti-inflammatory role in numerous autoimmune diseases, inflammatory diseases and inflammation-related tumors. However, the effect and mechanism of SOCS3 on vein graft disease is unclear. The purpose of this study was to investigate the effects of SOCS3 on the inflammation, migration and proliferation of VSMCs in vitro and the mechanism. The small interference RNA plasmid targeting rat SOCS3 (SiRNA-rSOCS3) and the recombinant adenovirus vector carrying rat SOCS3 gene (pYrAd-rSOCS3) were constructed, and the empty plamid (SiRNA-control) and vector (pYrAd-GFP) only carrying GFP reported gene were constructed as control. The rat VSMCs were cultured. There were two large groups of A (SOCS3 up-regulated): control group, IL-6/IFN-γ group, IL-6/IFN-γ+pYrAd-rSOCS3 group, IL-6/IFN-γ(+)pYrAd-GFP group; and B (SOCS3 down-regulated): control group, IL-6/IFN-γ group, IL-6/IFN-γ+SiRNA-rSOCS3 group and IL-6/ IFN -γ+SiRNA-control group. The pYrAd-rSOCS3 and SiRNA-rSOCS3 were transfected into VSMCs induced by IL-6/IFN-γ. After 24 h, real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to detect the mRNA and protein expression of SOCS3, STAT3 (only by Western blotting), P-STAT3 (only by Western blotting), IL-1β, IL-6, TNF-α, MCP-1 and ICAM-1. The MTT, Transwell assay and flow cytometry were used to examine VSMCs proliferation, migration and cell cycle progression, respectively. As compared with control group, the mRNA and protein expression of SOCS3, STAT3, P-STAT3, IL-1β, IL-6, TNF-α, MCP-1 and ICAM-1 was significantly up-regulated in VSMCs stimulated by IL-6/IFN-γ. However, in VSMCs transfected with pYrAd-rSOCS3 before stimulation with IL-6/IFN-γ, the expression of SOCS3 mRNA and protein was further up-regulated, and that of STAT3, P-STAT3, IL-1β, IL-6, TNF-α, MCP-1 and ICAM-1 was significantly down-regulated as compared with IL-6/IFN-γ group and IL-6/IFN-γ+pYrAd-GFP group. The expression of those related-cytokines in IL-6/IFN-γ+SiRNA-rSOCS3 group was markedly increased as compared with IL-6/IFN-γ group and IL-6/IFN-γ+SiRNA-control group. The absorbance (A) values, the number of cells migrating to the lower chamber, and percentage of cells in the G2/M+S phase were increased in VSMCs stimulated by IL-6/IFN-γ. In VSMCs incubated with pYrAd-rSOCS3 or SiRNA-rSOCS3 before IL-6/IFN-γ stimulation, the A values, the number of cells migrating to the lower chamber, and the percentage of cells in the G2/M+S phase were significantly decreased, and increased respectively. These results imply that IL-6/IFN-γ, strong inflammatory stimulators, can promote transformation of VSMCs phenotype form a quiescent contractile state to a synthetic state by activating JAK2/STAT3 pathway. Over-expresssed SOCS3 might inhibit pro-inflammatory effect, migration and growth of VSMCs by blocking STAT3 activation and phosphorylation. These data in vitro confirm that SOCS3 may play a negatively regulatory role in development and progression of vein graft failure. These conclusions can provide a novel strategy for clinical treatment of vein graft diseases and a new theoretic clue for related drug development.
Animals ; Blotting, Western ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Cytokines ; genetics ; metabolism ; Flow Cytometry ; Gene Expression ; Inflammation Mediators ; metabolism ; Interferon-gamma ; genetics ; metabolism ; pharmacology ; Interleukin-6 ; genetics ; metabolism ; pharmacology ; Janus Kinase 2 ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Phosphorylation ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; STAT3 Transcription Factor ; genetics ; metabolism ; Signal Transduction ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; genetics ; metabolism

The historical evolution， fermentation technology and key links of Sojae Semen Praeparatum （SSP） were sorted out by consulting ancient books and modern literature， and the influencing factors and control methods of quality were analyzed and summarized in order to provide reference for the quality control of SSP. After analysis， it was found that in the fermentation process of SSP， fermentation strains， miscellaneous bacteria， temperature and humidity were all important factors affecting the quality of SSP. The condition control of "post fermentation" process has been paid more attention to in the past dynasties. In addition， the delicious SSP recognized in ancient times should be made from mold fermentation， and the breeding and application of fermented mold may be the key point to solve the quality problem of SSP. Therefore， based on the evaluation indexes of SSP in the past dynasties， it is of great significance to study and optimize the technological conditions such as strain， temperature and humidity in depth to improve the quality of SSP.

Based on the systematic retrieval and the reported components of Sojae Semen Nigrum and Sojae Semen Praeparatum, this study conducted in-depth analysis of conversion of components in the fermentation process, and discussed types and possible mec-hanisms of conversion of chemical components, so as to provide the basis for studying technology, medicinal ingredients and quality standards. According to the analysis, there is a certain degree of conversion of nutrients(like protein, sugar, lipid), bioactive substances(like isoflavones, saponins, γ-aminobutyric acid) and other substances(like nucleosides, melanoids, biamines, etc) in the process of fermentation.
Chromatography, High Pressure Liquid ; Fermentation ; Isoflavones/analysis* ; Semen/chemistry* ; Soybeans