OBJECTIVETo investigate the regulation trend of Jinxin Oral Liquid (JXOL) on the expression of negative regulatory factor of TLR3 signaling pathway SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points.

METHODSTotally 75 BALB/c mice were randomly divided into 5 groups, i.e., the normal control group, the model group, the ribavirin group, the high dose JXOL group, and the equivalent dose JXOL group, 15 in each group. Each group had 3 intervention ways (I, II, and III) with 5 mice treated in each group. BALB/c mice were nasally infected with respiratory syncytial virus (RSV), and treated by different intervention ways. After intervention, mice were killed and their lung tissues were sampled, mRNA expression levels of RSV-M, SOCS1, and IFN-β were detected by Real time PCR. The expression of SOCSl at the protein level was detected by Western blot.

RESULTSCompared with the normal control group, the mRNA expression level of SOCS1 and IFN-β, and the protein expression level of SOCS1 increased significantly in the model group intervened by intervention I and II (all P < 0.01), but the mRNA expression level of IFN-β decreased significantly in model group intervened by intervention III (P < 0.01). Compared with the model group, the mRNA expression level of RSV-M all significantly decreased in the high dose JXOL group and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and III and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01). The mRNA expression level of IFN-β significantly decreased in the high dose JXOL group intervened by intervention I and II and the equivalent dose JXOL group intervened by intervention I (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III and the equivalent dose JXOL group intervened by intervention III (all P < 0.01). The protein expression level of SOCS1 significantly decreased in the high dose JXOL group intervened by intervention I and the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01), while it significantly increased in the high dose JXOL group intervened by intervention III (all P < 0.01). Compared with the high dose JXOL group, the mRNA expression level of RSV-M decreased significantly in the equivalent dose JXOL group intervened by intervention I and II (P < 0.01). The mRNA expression level of SOCS1 and IFN-β decreased significantly in the equivalent dose JXOL group intervened by intervention I (P < 0.01), but the mRNA expression level of IFN-β increased significantly in the equivalent dose JXOL group intervened by intervention II and III (all P < 0.01). The protein expression level of SOCS1 decreased significantly in the equivalent dose JXOL group intervened by 3 intervention ways (all P < 0.01).

CONCLUSIONSJXOL could inhibit the expression of SOCS1 in the lung tissue of RSV infected BALB/c mice at different time points. Its regulatory effect might be associated with promoting the expression of interferon type I and further fighting against RSV.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Lung ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; Respiratory Syncytial Virus Infections ; drug therapy ; metabolism ; Respiratory Syncytial Viruses ; Ribavirin ; Signal Transduction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism ; Toll-Like Receptor 3 ; metabolism

OBJECTIVETo evaluate the clinical effects of single posterior debridement, bone grafting, internal fixation and local chemotherapy in treating thoracolumbar spinal tuberculosis.

METHODSFrom February 2009 to September 2012,11 patients with thoracolumbar spinal tuberculosis were treated by single posterior debridement, bone grafting, internal fixation and local chemotherapy. There were 7 males and 4 females, aged from 27 to 65 years old with an average of 53.7 years. The courses of disease was from 3 months to 2 years with the mean of 9 months. According to ASIA standard of spinal cord injury, 3 cases were grade C and 8 cases D. After treatment, clinical effects were evaluated by ASIA grade, visual analogue score (VAS) and Oswestry Disability Index (ODI); kyphosis Cobb angle change was observed by X-rays.

RESULTSEleven patients were followed up from 12 to 29 months with an average of 18 months. ASIA grade of spinal cord injury, 3 patients with grade C improved to grade D in 2 cases and grade E in 1 case 8 patients with grade D improved to grade E in 7 cases and unchanged in 1 case. VAS decreased from preoperative 6.10 ± 1.30 to 1.70 ± 0.80 at 3 d after operation (P < 0.05). ODI improved from preoperative (68.36 ± 10.41)% to (14.55 ± 8.99)% (P < 0.05) at 3 d after operation. Kyphotic Cobb angle was corrected from preoperative (22.64 ± 4.84)° to (4.27 ± 1.49)° (P < 0.05) on the 3rd day after operation, and angle loss was mild at final follow-up, there was no significant difference between postoperative at 3 d and final follow-up.

CONCLUSIONSingle posterior debridement, bone grafting, internal fixation and local chemotherapy for the treatment of thoracolumbar spinal tuberculosis can effectively remove the lesion, improve nerve function and correct deformity, has advantage of single incision, little trauma, and low recurrence rate. But it still need long-term and systemic treatment with anti-TB drugs.

Adult ; Aged ; Bone Transplantation ; Debridement ; Female ; Humans ; Internal Fixators ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; therapy

OBJECTIVETo evaluate the midterm follow-up results of extended release of posterior clearance in total knee arthroplasty.

METHODSA total of 120 patients with knee osteoarthritis were equally randomly assigned to the experimental group and control group, and underwent unilateral TKA from March 2010 to March 2012. In experimental group, there were 21 males and 39 females with an average age of (62.2±10.9) years old. In the control group, there were 25 males and 35 females with an average age of (64.9±11.4) years old. All the patients were performed using the anterior knee approach. During operation, after osteotomy of the tibia and the femoral condyle, extended release of the posterior knee clearance were taken in experimental group, while only the clearance of osteophyte in the posterior condyle were performed in the control group. The KSS scores including knee functional score and knee clinical score,as well as the range of motion (ROM) of patients, were compared between the two groups at midterm follow-up.

RESULTSTotally 49 patients in the experimental group and 54 patients in the control group were followed up, and the median follow-up time was 46 months. The knee functional score of patients in the experimental group was 91.3±3.4, which was better than 86.4±3.9 of patients in the control group; initiative ROM of flexion of patients in the experimental group was (133.2±5.9)°, which was better than (126.9±7.4)° of patients in the control group. There were no significant difference of knee clinical score between 86.9±4.6 of patients in the experimental group and 85.7±5.1 of patients in the control group, and the initiative ROM of extension between (0.5±1.1)° and (0.3±1.2)°.

CONCLUSIONExtended release of the posterior knee clearance contributes to the knee function and initiative flexion ROM during a midterm follow-up and patients benefit.

Aged ; Arthroplasty, Replacement, Knee ; methods ; Case-Control Studies ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; surgery ; Range of Motion, Articular

OBJECTIVETo observe the effect of isolating the chondrocytes from articular cartilage with the method of three-step enzymatic digestion, and the biological characteristics of the isolated chondrocytes during cultivation in vitro in order to evaluate their biological activity.

METHODSThe method of three-step enzymatic digestion was designed that the articular cartilage was digested one by one with the 1 g/L trypsin and 1 g/L EDTA, 1 g/L hyaluronidase and 2 g/L collagenase I in the culture medium to isolate chondrocytes. The harvesting and viability rate of the primary chondrocytes were detected. During the passage cultivation in vitro, the changes of the chondrocytes shape and growth were observed, the changes of the collagen type I and II and aggrecan in the extracellular matrix were investigated and detected.

RESULTS(1) The extracellular matrix of articular cartilage was completely dissolved by the three-step enzymatic digestion, and the chondrocytes were completely isolated from the solid matrix. The number of the harvested chondrocytes from every gram of wet cartilage was 50.3 x 10(6) on average, and their viability rate was 98.8% on average. (2) The primary and first passage chondrocytes had triangle or multi-angle shape, and became elliptic shape at the growing confluence with the positive immunohistochemical stain of collagen type II and the strong heterochromia to toluidine blue. The content of sulfate glycosaminoglycans (GAG) in the extracellular matrix of the primary passage cells was (92 +/- 10) microg/cm(2). The chondrocytes after the third passaging gradually became spindle shape with the negative stain of collagen type II and the weak heterochromia to toluidine blue. The content of sulfate GAG of the fourth passage cells was (48 +/- 12) microg/cm(2).

CONCLUSION(1) The method of three-step enzymatic digestion can make the extracellular matrix of articular cartilage completely degraded, and has advantages in the high efficiency of harvesting primary chondrocytes, high cellular viability rate and simple manipulation. (2) The primary and first passage chondrocytes have fine biological activity, and the chondrocytes after the third passaging have lost their special biological activity.

Animals ; Cartilage, Articular ; cytology ; drug effects ; Cell Culture Techniques ; Cell Separation ; methods ; Cells, Cultured ; Chondrocytes ; cytology ; Collagenases ; pharmacology ; Female ; Hyaluronoglucosaminidase ; pharmacology ; Male ; Rabbits ; Trypsin ; pharmacology

OBJECTIVETo optimize primarily acupuncture program for dysphagia after cerebral infarction.

METHODSSixty-three patients with pseudobulbar palsy dysphasia after cerebral infarction were selected as the subjects. Water swallow test was adopted for observation index. Orthogonal design was used to study the different combinations of treatment programs among four factors and three levels, named acupuncture opportunity (factor A: A1: 1-10 d, A2: 11-20 d, A3: 21-40 d), point combination (factor B: B1: points in the neck, B2: points in the neck + points in the glossopharyngeal part, B3: points in the neck + points in the glossopharyngeal part + differentiating point), needling depth (factor C: C1: shallow puncture, C2: moderate puncture, C3: deep puncture), and treatment session (factor D: D1: 2 sessions, D2: 4 sessions, D3: 6 sessions); as well as the influences on pseudobulbar palsy dysphagia after cerebral infarction so as to determine primarily the optimized acupuncture program for dysphagia after cerebral infarction.

RESULTSThe direct analysis of orthogonal design indicated that the best level and the combination of four factors and three levels were A1 (or A2), B3, C3 and D3. It meant that for the patients less than 20 days for sickness, the points in the neck, points in the glossopharyngeal part and points selected according to syndrome differentiation were selected for deep puncture and the treatment lasted for 6 sessions. This program achieved the best improvement in water swallow test. The comparison between the primary effect and secondary effect among four factors (variance analysis) showed that factor A and C were the significant factors (P < 0.05), factor B and D were not significant (P > 0.05). It was suggested that acupuncture opportunity and needling depth played the significant roles in clinical efficacy. The comparison among different levels in terms of the significant factors A and C (multiple comparison) presented that A1 and A2 were superiorly significant than A3 (P < 0.001), A1 was not different in significance from A2 (P > 0.05). C3 was superiorly significant than C2 and C1 (P < 0.001, P < 0.01) and C2 was superiorly significant than C1 (P < 0.05).

CONCLUSIONBy the comprehensive summarization of direct analysis, variance analysis and multiple comparison combined with actual clinical situation, the optimal program of acupuncture treatment for pseudobulbar palsy dysphasia after cerebral infarction is that in terms of the four factors and three levels, for the patients within 1 to 20 days for sickness, acupuncture is applied to the points in the neck or combined with points in the glossopbaryngeal part or the points se-i lected according to syndrome differentiation, with deep puncture, for 4 to 6 sessions of treatment.

Acupuncture Therapy ; Aged ; Aged, 80 and over ; Cerebral Infarction ; complications ; Deglutition ; Deglutition Disorders ; etiology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo study the ultrastructure and Ca/P element spectrometry of distraction osteogenesis (DO) for reconstruction of cleft palate (CP), so as to explore the osteogenesis and remodeling of new bone in situ.

METHODS23 rhesus macaques were operated to establish animal models of CP. 2 monkeys didn't received DO as controls. The other 21 monkeys in experimental group underwent DO to correct both bony and soft tissue defects in palate. The distraction was performed at a rate of 0.8 mm/d, twice a day until the cleft was closed. After fixation for 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were sacrificed to get the specimens at the distraction gap. The scanning electron microscopic study and Ca, P elements spectrometric analysis were adopted. There were also two unoperated animals as sham group.

RESULTSAfter fixation for 1-2 weeks, the distraction gap was full of collagen fibers oriented along vector of distraction. Few trabeculae was seen at the margin area. After fixation for 4-6 weeks, active osteogenesis was presented with new formed bone trabeculae and abundant cellular component. After fixation for 8-12 weeks, the new formed bone became mature and couldn't distinguish from the normal bone. 24 weeks later, the bone between the distraction gap had a similar structure to the normal bone. Elements spectrometric analysis results indicated that in early stage of osteogenesis, the P and S peaks were relatively high while the Ca peak was much lower. During the late stage, the S peak was obviously decreased, and Ca/P ratio increased to normal level as in the empty control group.

CONCLUSIONSThe CP can be corrected by DO. The new bone between the distraction gap is formed and remodeled through intramembraneous osteogenesis.

Animals ; Cleft Palate ; metabolism ; pathology ; surgery ; Disease Models, Animal ; Female ; Macaca mulatta ; Male ; Microscopy, Electron, Scanning ; Osteogenesis ; Osteogenesis, Distraction ; methods ; Palate ; surgery ; ultrastructure

The total triterpene saponins of Psammosilene tunicoides have significant pharmacologic activity. Psammosilene tunicoides squalene synthase (PSS) is a gateway enzyme to regulate the biosynthesis of total triterpene saponins extracted from the root of Psammosilene tunicoides which is an endangered species. In this paper, cDNA encoding of PSS was cloned by the degenerate primer PCR and rapid-amplification of cDNA ends (RACE). The full-length of cDNA of PSS is 1663 bp, with an open reading frame (ORF) of 1 245 bp, encoding 414 amino acid polypeptide (calculated molecular mass, 47.69 kDa), 5'UTR (untranslated region) and 3'UTR are 260 bp and 158 bp, respectively. The deduced amino acid sequence of PSS has higher homology with the known squalene synthases of several species such as Panax notoginseng (83%), Panax ginseng (82%) and Glycyrrhiza glabra (82%) than that with Schizosacharomyces pombe (35%), Candida albicans (39%) and Homo sapiens (47%). The characterization of PSS was done by a series of methods, such as prokaryotic expression, the activity of enzyme in vitro, capillary gas chromatography (GC) and capillary gas chromatography mass spectrometry (GC-MS). The results showed that the cell-free extract of E. coli transformed with the recombinant plasmid can effectively convert farnesyl diphosphate into squalene in vitro. GenBank accession number is EF585250. Our research provided important base for the study of Psammosilene tunicoides secondary metabolism and metabolic engineering.
Caryophyllaceae ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Endangered Species ; Escherichia coli ; genetics ; metabolism ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Open Reading Frames ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; metabolism ; Sequence Homology, Amino Acid ; Transformation, Genetic

OBJECTIVETo investigate the mechanism of new bone formation in the distraction osteogenesis (DO) for correction of cleft palate (CP) in rhesus.

METHODSCP was created by operation in 23 rhesus. The CP was corrected with DO in 21 animals as experimental group. The distraction rate was 0.8 mm per day, two times a day. The bone fragments were fixed after cleft closure, every 3 animals were sacrificed to get specimen after 1, 2, 4, 6, 8, 12, 24 weeks of fixation. 6 days before sacrifice, tetracycline was administrated for labeling (30 mg/kg).

RESULTSThe hard and soft tissue def of fixation. At the same time, the bone volume and calcification between the distraction gap increased. The cleft in the control group could not b ect was successfully closed with DO by intramembrane osteogenesis. The new formed bone was remodeling and became maturation during the period e corrected spontaneously.

CONCLUSIONSThe DO can successfully correct both the soft and hard tissue defect in CP by intramembrane osteogenesis. The fixation is important for remodeling and maturation of the new formed bone.

Animals ; Biomarkers ; Cleft Palate ; pathology ; surgery ; Macaca ; Osteogenesis, Distraction ; Palate, Hard ; pathology ; Palate, Soft ; pathology

OBJECTIVETo study the mechanism of new bone formation and remodeling of distraction osteogenesis (DO) by analysis of the expression of osteopontin (OPN) and osteocalcin (OC).

METHODSRhesus were operated to reconstruct the animal model of cleft palate (CP). The CP was closed by DO in experimental group(n = 21). After consolidation of 1, 2, 4, 6, 8, 12, 24 weeks, every 3 animals were killed to collect the specimens, respectively. The OPN and OC and their mRNA were detected quantitatively by Real-time RT-PCR and ELISA, respectively. The animals in control group (n = 2) and sham group (n = 2) were used as control.

RESULTSThe mRNA expression of OPN increased since 2nd week of consolidation and reached the peak at 4th week (7.59 +/- 0.37). The mRNA expression of OC was up-regulated since 4th week, and reach the peak at 6th week (7.94 +/- 0.31). Then they decreased to about the level in sham group at 24th week (P > 0.05). The OPN and OC were highly expressed during 4 to 6 weeks of consolidation. During 8 to 12 weeks, they decreased like their mRNA expression.

CONCLUSIONThe intramembraneous new bone formation after DO can reconstruct the bone defect of CP. The new formed bone can be remodeled to be quite normal bone tissue.

Animals ; Cleft Palate ; metabolism ; surgery ; Macaca mulatta ; Osteocalcin ; metabolism ; Osteogenesis, Distraction ; Osteopontin ; metabolism

Objective To analyze the epidemic characteristics of hand-foot-mouth disease (HFMD) in Jiangsu province.Methods We downloaded the case-data of HFMD in Jiangsu province during 2009-2011 from the Chinese National Notifiable Infectious Disease Reporting System,and made a comprehensive analysis on the epidemiological features of it with descriptive epidemiological methods and retrospective space-time permutation scan statistics.Results A total of 285 414 cases were reported in Jiangsu,from 2009 to 2011,with an annual incidence of 122.66 per 100 000.There were 3686 severe cases in the 3 years accumulatively,accounting for 1.29％ of the total.Proportion of the cases being 5 years old or even younger was 93.64％.Scatteredly living children accounted for 64.08％ of the total cases and 78.65％ of the severe cases,respectively.The epidemics of HFMD were visible in each city of Jiangsu province,with a lowest annual incidence rate of 44.02 per 100 000 and a highest one up to 202.90 per 100 000.Regions as Suzhou,Nanjing,Wuxi had the highest incidence in the province,with cases in these three areas occupying almost 40％ of all.The numbers of severe cases in Suqian and in Yancheng cities increased by 339.22％ and 328.33％ in 2011 compared to in 2010,respectively,and the rates of increase in these two cities were much higher than those in the other regions.Two peaks of incidence were observed every year,with the highest occurring between April and June and the second occurring in November.The spatial-temporal distribution of HFMD was not random in Jiangsu province,from 2009 to 2011.Clusters for general cases in August and 7 clusters for severe cases were detected,respectively.12 359 cases of HFMD were laboratory confirmed in the said 3 years,including 10 414 common cases and 1945 severe cases.EV71 and CoxA16 accounted for 43.49％ and 37.07％ of common cases,respectively.In terms of the severe cases,the ratios were 80.82％ and 5.96％,respectively.Conclusion HFMD was highly endemic in Jiangsu province,and the situation of prevention and control for it is still grim.Scatteredly living children of 5 years old or younger were the major population at risk,and the epidemic in different regions and seasons was different.EV71 and CoxA16 were the major etiologic agents,but the etiologic constitute showed seasonal changes.