Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

OBJECTIVETo investigate the expression and relationship between HSP70 and caspase-3 in knee osteoarthritis.

METHODSForty adult SD rats were divided into experimental group and control group. Thirty rats in experimental group, anterior cruciate ligament transaction (ACLT) was cut off and partial meniscectomy of 1/3 inside incision were performed to reproduce knee osteoarthritis (KOA) model according to Hulth methos, and the other 10 rats was treated with nothing as control group. The rats were sent to the cage and free to move. At 1, 2 and 4 weeks later, the arthritis cartilage of femoral and tibial end were observed through immunohistochemistry staining and light microscope. Meanwhile, Mankin scale system was adopted for histomorphology evaluation.

RESULTSChanges of KOA such as hyperplastic synovium,erosion on the surface of cartilage and so on were found in experiment group, the expression of HSP70 was augmentation all the time, but the expression of caspase-3 was reduction 1 week later; no similar changes were found in control group. Mankin scale system showed that there were significant differences in the first week as compared with the second week and 4th week (both P < 0.01).

CONCLUSIONHeat shock protein inhibit the apoptosis of cartilage cells and protect the cartilage cells in knee osteoarthritis, the conservative treatment for clinical provide objective scientific basis.

Animals ; Apoptosis ; Cartilage ; pathology ; Caspase 3 ; physiology ; Chondrocytes ; pathology ; Female ; HSP70 Heat-Shock Proteins ; physiology ; Male ; Osteoarthritis, Knee ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate body weight and the body mass index (BMI) in girls with adolescent idiopathic scoliosis (AIS).

METHODSSix hundred and thirteen AIS female patients admitted from January 2005 to January 2007 and 449 healthy girls were recruited for this study with a range of age from 12 to 16 years old. The direct indexes included body height, body weight and date of birth. The indirect indexes included corrected body height, BMI and corrected BMI. The corrected body heights of AIS patients were adjusted by using the greatest Cobb angle to correct for spinal deformity according to Bjure's formula. Two-tailed Student's test was used for group comparison.

RESULTSIn the AIS patients, the maximum Cobb angle was (31 +/- 11) degrees (11 degrees to 77 degrees). The corrected body height were significantly higher in AIS patients than in the normal controls from 12 to 16 years old (P < 0.05). The body weight were significantly lower in AIS girls than in normal controls from 12 to 16 years of age (P < 0.01). From 12 to 16 years of age, the BMI were (17.6 +/- 1.9), (17.9 +/- 2.5), (17.9 +/- 2.1), (18.6 +/- 2.3) and (19.0 +/- 1.9) kg/m2 in AIS patients; while the BMI were (19.5 +/- 3.4), (19.8 +/- 3.0), (20.4 +/- 2.9), (20.4 +/- 2.8) and (20.2 +/- 2.2) kg/m2 in normal controls. The BMI were significantly lower in AIS girls than the normal controls from 12 to 16 years of age (P < 0.001).

CONCLUSIONSAfter the onset of puberty, significantly longer corrected height, lower body weight and lower BMI were found in AIS patients. Results of this large-scale study revealed the presence of abnormal growth in AIS patients during peripubertal development.

Adolescent ; Body Height ; Body Mass Index ; Body Weight ; Child ; Female ; Humans ; Kyphosis ; physiopathology ; Scoliosis ; physiopathology

OBJECTIVETo investigate the association between the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene polymorphisms with the predisposition and disease severity of thoracic adolescent idiopathic scoliosis (AIS).

METHODSThree hundred and fifty-four female thoracic AIS patients treated from January 2007 to March 2009 and 210 healthy female who underwent health examination during March 2005 to June 2006 as normal controls were recruited in this study. One SNP-418G/C (rs8179090) in the promoter region were selected for TIMP-2 gene. PCR- RFLP was used for genotyping.

RESULTSNo significant differences of genotype and allele frequency distribution were found between AIS patients and normal controls (P > 0.05). In AIS patients, the frequency of C allele of the patients with the body mass index (BMI) < 17 kg/m(2) was significantly higher than those with the BMI > or = 17 kg/m(2) (P < 0.05), and the frequency of C allele of cases with the major Cobb angle > or = 40 degrees was significantly higher than that with Cobb angle < 40 degrees (P < 0.05). Among the patients who reached skeletal maturity without any interference of natural history, significantly higher average maximum Cobb angle was found in patients with GC and CC genotype compare with those with GG genotype.

CONCLUSIONSThe SNP-418G/C (rs8179090) in the promoter region of TIMP-2 gene may be associated with abnormal growth pattern and curve progression of thoracic AIS. TIMP-2 gene is a disease-modifier gene of thoracic AIS.

Adolescent ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Scoliosis ; genetics ; Tissue Inhibitor of Metalloproteinase-2 ; genetics

OBJECTIVETo study the relationship between T1 tilt and cosmetic shoulder balance in adolescent idiopathic scoliosis (AIS) patients.

METHODSSeventy-one Lenke type 2 AIS patients were recruited into the present study from January 2010 to December 2011. There were 61 female and 10 male patients, the average age was (15.1 ± 2.9) years (range 10-18 years); the average Risser sign was 2.8 (range 1-5). The patients were photographed from the back in neutral standing position on level ground wearing underpants. Also, all the patients had a standing posterior-anterior radiograph in a relaxed standing position. The cosmetic shoulder height (CSH), which included cosmetic inner shoulder height (CSHi) and cosmetic outer shoulder height (CSHo), were measured in the photographs. Positive value was defined as left shoulder was higher than the right shoulder, and negative value was defined as right shoulder was higher than the left shoulder. CSH > 5 mm was defined as positive CSH, CSH < -5 mm was defined as negative CSH, -5 mm ≤ CSH ≤ 5 mm was defined as leveled CSH. T1 tilts were measured in the posterior-anterior radiographs. Positive value was defined as the left proximal vertebral body up and right lower vertebral body down, and negative value was defined as the right proximal vertebral body up and left lower vertebral body down. T1 tilt > 5° was defined as positive T1 tilt, T1 tilt < -5° was defined as negative T1 tilt, -5° ≤ T1 tilt ≤ 5° was defined as leveled T1 tilt.

RESULTST1 tilt was found to be significantly correlated with CSHi and CSHo (r = 0.25 and 0.28, P < 0.05).For positive T1 tilt patients, there were 59.0% (36/61) patients with positive CSHo, 37.7% (23/61)with leveled CSHo, and 3.3% (2/61)with negative CSHo.For the patients with leveled T1 tilt, it was 3/10, 5/10 and 2/10.For positive T1 tilt patients, there were 83.6% (51/61) patients with positive CSHi, 11.5% (7/61) with leveled CSHi, and 4.9% (3/61) with negative CSHi; For the patients with leveled T1 tilt, it was 6/10, 2/10 and 2/10.

CONCLUSIONSAlthough positive correlation is found between T1 tilt and cosmetic shoulder balance, positive T1 tilt is not an indicator of higher left shoulder.Elevated left shoulder, leveled shoulders and elevated right shoulder are all found in positive T1 tilt patients.

Adolescent ; Child ; Esthetics ; Female ; Humans ; Male ; Postural Balance ; Radiography ; Scoliosis ; diagnostic imaging ; Shoulder ; anatomy & histology ; Thoracic Vertebrae ; anatomy & histology

BACKGROUNDDouble plate osteosynthesis is the standard treatment for intra-articular fractures of the distal humerus. However, there is still controversy concerning dual plate positions in terms of providing optimal stability. The purpose of this retrospective study was to compare the clinical outcomes in patients with type C intra-articular distal humeral fractures using perpendicular and parallel plating methods.

METHODSBetween March 2008 and June 2011, a total of 45 patients with type C distal humerus fractures were treated using two different dual plating methods. Of them, 24 patients were treated by perpendicular plating (group I) and 21 patients were treated by parallel plating (group II). The surgical time, blood loss, and union time were compared between the two groups. The flexion-extension arc, the total range of flexion and extension at the end of follow-up were compared between the two groups. The Mayo Elbow Performance Score (MEPS) was used to determine the elbow functional results.

RESULTSAll patients were followed up. The mean duration of follow-up was 16 months (range 12 - 25 months) in group I and 15.5 months in group II (range 12 - 25 months). There were no significant differences in the surgical time, blood loss, and the bone union time between the two groups. In group I, the mean elbow flexion-extension arc was 101° and the mean MEPS was 85 points. The rate of excellent and good results was 87.5%. In group II, the mean flexion-extension arc was 100° and the mean MEPS was 86.1 points. The rate of excellent and good results was 90.5%. There were no significant differences in the MEPS, flexion-extension arc, and the total range of flexion and extension between the two groups.

CONCLUSIONSPerpendicular and parallel plate configurations with the appropriate surgical techniques can provide anatomical reconstruction and stable fixation of type C intra-articular distal humeral fractures and allow early mobilization of the elbow after an operation. The occurrence of post-operative elbow stiffness can be reduced and good outcomes can be obtained.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Retrospective Studies

According to the 2020 world cancer data report， the prevalence of prostate cancer ranks second in male malignant tumors and the mortality fifth. In China， due to the poor living and eating habits， the prevalence of prostate cancer is still rising， and advanced prostate cancer can adapt to and tolerate androgen castration related drugs through a variety of mechanisms， which often indicates poor therapeutic outcomes in the late stage. Houttuyniae Herba， a medicinal plant of family Saururaceae， has been officially identified both as food and medicine by the National Health Commission. The existing studies have shown that Houttuyniae Herba inhibits the proliferation， migration， and invasion of prostate cancer and induces its apoptosis and cell cycle arrest via multiple targets， links， and pathways. These anti-tumor activities can also be observed in breast cancer， leukemia， gastric cancer， colorectal cancer， and other tumor cells. Such activities are mainly related to the inhibited expression of cyclin and cyclin-dependent kinase， regulation of apoptosis-related proteins， activation of apoptosis-related pathways， reduced secretion of matrix metalloproteinase， inhibition of activator of transcription 3 （STAT3）/Zinc finger protein （Snail） / transcription factor twist signaling pathway， and antagonization of growth factor receptor， androgen， inflammation， and oxidative stress in promoting the occurrence and development of prostate cancer. In addition， Houttuyniae Herba is able to alleviate diabetes， regulate blood glucose-lowering pathways， and boost immunity by inhibiting the protein kinase B （Akt）/mammalian target of rapamycin （mTOR） and mitogen-activated protein kinase （MAPK） signaling pathways， thus suppressing the occurrence and progression of prostatic cancer. In conclusion，Houttuyniae Herba can be used as a potential drug against prostate cancer.

OBJECTIVETo investigate the effect of sympathectomy on the development and progression of scoliosis in bipedal C57BL/6J mice model.

METHODSSixty female 3-week-old C57BL/6J mice were selected to establish bipedal scoliotic mice model after amputations of forelimbs and tails. All mice were randomly divided into three groups, 20 mice for each group. Group 1 received daily intraperitoneal injection of 0.9% saline (5 mg/kg); while Group 2 and 3 received sympathectomy by daily intraperitoneal injection of propranolol (20 mg/kg) and guanethidine sulfate (40 mg/kg), respectively. Posteroanterior X-rays were obtained at 20th week. Curves were measured using Cobb method and scoliosis was defined as a Cobb angle of > 10°. Incidence of scoliosis and severity of curves were compared among groups using Chi-square test and One-way analysis of variance, respectively.

RESULTSThere were 17 (85.0%) mice presented scoliosis in Group 1; whereas 11 (55.0%) and 10 (50.0%) mice presented scoliosis in Group 2 and 3, respectively. The incidence of scoliosis was found to be higher in Group 1, and the difference was statistically significant (χ(2) = 6.172, P = 0.046). As for curve magnitudes, the mean Cobb angle was 20° ± 9° in Group 1, 10° ± 7° in Group 2, and 12° ± 8° in Group 3. The mean Cobb angle of Group 1 was significantly greater than those of Group 2 and 3 (F = 9.545, P < 0.001), but there was no significant difference in mean Cobb angle between Group 2 and 3.

CONCLUSIONSSympathetic nervous system may be involved in the development and progression of scoliosis in bipedal C57BL/6J mice model. Sympathectomy do not seem to dramatically decrease the incidence of scoliosis, probably due to that bipedalism itself may also be a cause of scoliosis in this animal model.

Animals ; Disease Models, Animal ; Female ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Scoliosis ; pathology ; Sympathectomy ; Sympathetic Nervous System ; physiopathology ; Thoracic Vertebrae