Objective To comprehand and grasp epidemical distribution characteristics of colorectal cancer cases. WT5”HZMethods Between 1975 and 1999, 1 145 cases of colorectal carcinoma who had undergone a surgery in Nanfang Hospital were retrospectively studied and a survey of clinical epidemical distribution characteristics was made. ResultsWT5”BZ ①Though more elder cases were found in recent years, young patients still made up a high proportion(19%) of the cases.②The occurrence of mucoid and signet-ring cell carcinoma in young cases was higher than that in the elders( P 0 05). KG2Conclusion The study of clinical epidemiology provided dependabal bases for prevention and therapy of colorectal carcinoma.

Human height, a complex trait with over 80% heritability, is determined by genetic and environment factors. Currently, a certain amount of specific variants about human height have been found, especially with the widely used of genome-wide association studies (GWAS) in genetics research, making it a great progress for the discovering of height associated genes. However, single nucleotide polymorphisms (SNPs) found by GWAS can only explain a minority of the heritable. This article reviews the progress of GWAS about height associated genes and missing heritability domestic and overseas.

Objective To improve the tri-primer allele gene amplification for realizing the single nucleotide polymorphisms (SNP) genotyping of the peripheral blood sample .Methods Aiming at the peripheral blood samples with the clinical usual antico-agulation processing by EDTA ,heparin and citrate ,with the locus rs1165205 as the target site ,the buffer solution(YW) suitable for whole blood was prepared and the PCR amplification system and the amplification condition were optimized for realizing the detec-tion of SNP genotyping .Results The genotyping results of locus rs1165205 by improved tri-primer allele gene amplification method were consistent with the results of the Sanger sequencing method ,and the peripheral blood samples treated by different anticoagu-lant were genotyped by the improved tri-primer ASA .Among 80 samples ,various genotypes of locus rs1165205 had no statistical differences in the distribution between the gout population and non-gout population(P= 0 .335) .Conclusion The improved tri-primer allele gene amplification method can be adopted to conduct the rapid genotyping research on gout SNP locus of the peripheral blood samples with the clinical usual anticoagulation processing .

Objective To investigate the operation key points, instrument improvement and shortterm effects in total en bloc spondylectomy (TES) via a single posterior approach for thoracic and lumbar tumors. Methods A series of modified instruments have been designed for the TES, including threadwire saw (T-saw) with a diameter of 0.81 mm, director and clamping for the saw, L shape and furcation osteotomes.The corpectomy of original TES which was defined as "one step dissection" from anteriorly to posteriorly, was modified into "two step dissection" which means that corpectomy was performed with saw cutting anteriorposteriorly and the L shape cutting posterior-anteriorly. In the cases with difficulty in pediculotomy using a T-saw, furcation osteotome was used for pediculotomy. Ten patients with thoracic or lumbar tumors were treated with the modified TES. There were 1 case of bone giant cell tumor, 1 case of bone neurilemmoma and 8 cases of metastatic tumors. All patients suffered moderate-severe pain and neurological deficit. Results The average follow-up period was 8.1(3.3-18.1) months. The average operating time was 7.8 h(6.0-10.3 h),and average blood loss was 2100 ml (1200-3500 ml). No disruption of dural mater, the leakage of cerebrospinal fluid, iatrogenic spinal cord injury and major vessel damage occurred. Two patients who underwent pleura disruption happened during the operation were treated with intrathoracic drain remedy. Among 7 cases with thoracic tumors, significant improvement in neurological function were achieved in 5 patients with the improvement of one grade in ASIA classification, while no change was found in 2 cases. In 3 cases with lumbar tumor, lumbar nerve root pain relieved and the muscle strength had recovered to grade 4 at least postoperatively. Conclusion Significant improvement has been achieved in the maneuverability and safety of the modified surgical techniques in TES with a single posterior approach for thoracic and lumbar tumors.

Objective To investigate the perioperative risk of deep vein thrombosis (DVT) in patients with hepatic cirrhosis that underwent total hip arthroplasty (THA), and to evaluate the safety and feasibility of individualized anti-coagulation treatment.Methods There were 25 patients complicating hepatic cirrhosis that underwent THA (from Jan.to Dec.2014), including 17 males and 8 females, aged 57.9t9.2 years.The primary causes of THA were avascular necrosis of the femoral head (eighteen cases) and osteoarthritis of the hip (seven cases).Low molecular weight heparin (LMWH) was applied for anti-coagulation treatment.Parameters of hepatic function and coagulation function of THA cases (randomized thirty cases, from Jan.2008 to Dec.2008) without hepatic cirrhosis were used as reference for monitoring.For the cases of massive blood loss or upper gastrointestinal hemorrhage, a LMWH administration pause and an administration of fresh frozen plasma and clotting factors were performed in order to maintain a hemorrage/coagulation balance.The clinical outcome of the hip joint was evaluated and complications were treated.A subsequent follow-up was also carried out after perioperative period.Results All cases received successful surgeries and followed up.The follow-up duration was 34± 15.7 months.The preoperative Harris hip score was 32.4± 10.2 points, while the most recent follow-up score was 82.9±6.1 points, which was statistically significant.Dislocation, periprosthetic fracture and periprosthetic infection were absent.All cases received individualized anti-coagulation treatments during peripoerative period.A hemorrage/coagulation balance was achieved.The dynamic parameter curves did not present excessive deviation from reference.One case encountered intermuscular hematoma of the lower limbs 48 hours postoperatively, which was solved by a LMWH pause and administration of fresh frozen plasma and clotting factors.One case suffered upper gastrointestinal hemorrhage five days postoperatively, which was controlled by a LMWH pause and the administration of somatostatin and proton pump inhibitor.Jaundic got worse in one case three days postoperatively but got relieved after treatment.Overt blood loss was 686t141.8 ml.Perioperative death, hepatic failure, hepatic encephalopath, hepatorenal syndrome were absent.No DVT was observed.Conclusion There are risks of DVT in patients of hepatic cirrhosis.Individualized anti-coagulation treatment is needed during perioperative period of THA.

[Objective]This study was designed to construct a recombinant adenovirus vector contains LMP-1 gene,and investigate the osteoinductive activity of MSC which were transfected recombinated adenoviral vector carrying LMP-1 gene.[Methods]Total RNA was extracted from mt osteoblast and the LMP-1 gene was acquired by RT-PCR,the LMP-1 gene and entry vector pENTR/D-TOPO were used to create the entry clone with the directional TOPO clone technology,then the entry clone and the expression vector were used to create the expression clone throush the LR recombination reaction.The adenovirus expression clone was linearized by PacI and transfected to the 293A cell line to harvest a high titer.Ad-LMP-1 was infected into the 3rd passage MSC,the expression of LMP-1 was detected by Western blot.The osteogenic activity of MSC was evaluated by the expression of collagen Ⅰ,ALP,osteocalcin and the formation of bone nodule.[Result]The LMP-1 gene was successfully acquired and confirmed,the entry clone and the expression clone were both verified by enzymes digestion,and the expression clone was further confirmed by sequenced.The expression of LMP-1 was detected successfully in MSC.The increasing expression of collagen Ⅰ,osteocalcin.ALP and bone nodule were observed by comparing to the control group.[Conclusion]Gateway technology not only make construction of the pAd-LMP-1 recombination adenovirus vector simple and fast,but also get a high transfection efficacy in MSC.LMP-1 gene can induce the osteoblast differention of MSCs,and improve its osteogenic activity.The adenovirus vector is reliable to be used in further gene therapy research.

BACKGROUND:How to obtain sufficient autogeneic chondrocytes is a problem which must be answered as soon as possible in both the transplantation of chondrocytes and the development of cartilage engineering.Bone marrow-derived mesenchymal stem cells have the potential of multidirectional differentiation. Under different induced conditions, they can differentiate into multiple tissue cells, such as chondrocyte, osteoblasts , sarcoblast, nerve cells and so on. Insulin-like growth factor-I plays an important role in regulating the formation and development of limb and cartilage.OBJECTIVE:To observe the effect of the insulin-like growth factor-I and culture solution of chondrocyte on inducing bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes in vitro.DESIGN:Open experiment.MATERIALS:This experiment was carried out at the Medical Study Center, Second Hospital Affiliated to Sun Yat-sen University from March to November 2003.Human bone marrow-derived mesenchymal stem cells were harvested from 4 to 6-month old embryos, all of which were from pregnant women who needed to terminate of pregnancy by induction delivery with water bag for health.METHODS:Human embryonic mesenchymal stem cells were isolated with Percoll separating medium. Subsequently, the cells were amplified in vitro,and the expression of surface makers of bone marrow-derived mesenchymal stem cells, such as CD44, CD71, CD34 and CD45 were measured with flow cytometer to identify the cells in our experiment. 100 μg/L insulin-like growth factors and culture solution of chondrocytes were added in the culture medium of bone marrow-derived mesenchymal stem cells of the fourth generation. The morphological changes of induced cells were observed with an inverted microscope. The expression of type Ⅱ cartilage matrix was observed by collogen immunohistochemistry. The proteoglycan level in the cells was detected, too.MAIN OUTCOME MEASURES:Phenotype of bone marrow-derived mesenchymal stem cells was identified through detecting the expressions of CD34, CD44 and CD4.Type Ⅱ collagen immunohistochemistry and the change of cellular ability to secrete proteoglycan after induction were observed to determine whether bone marrow-derived mesenchymal stem cells can differentiate into chondrocytes.RESULTS: ① Being observed under an inverted microscope,the bone marrow-derived mesenchymal stem cells presented a morphology like fibroblasts when they were cultured in vitro. ②Identification of surface antigen of bone marrow-derived mesenchymal stem cells: These cells were detected to have good homogenicity with flow cytometer. The fourth generation of bone marrow-derived stem cells positively expressed CD44, negatively expressed CD34 and CD45, suggesting these cells had the characteristics of bone marrow-derived mesenchymal stem cells. ③Observation of the morphological change of chondrocytes induced by bone marrow-derived mesenchymal stem cells under optical microscope: chondrocyte condition culture solution and insulin-like growth factors-Ⅰ were added to co-culture.During the process of culture, bone marrow-derived mesenchymal stem cells were seen to have a shape of round gradually. Fifteen days later, some cells presented a shape of short-shuttle or polygon with short mutations,which were the shape characteristics of chondrocytes. ④Immunohistochemical staining of Type Ⅱ collagen: In the insulin-like growth factor group,72.5% cells had many brown granules in cytoplasm, which were weakly positive or strongly positive expression of Type Ⅱ collagen. In the control group, bone marrow-derived mesenchymal stem cells was negative expression of type Ⅱ collagen on the 15th day. ⑤ Measurement of proteoglycan level: After co-culture with insulin-like growth factor-Ⅰ and chondrocyte culture solution for 15 days, proteoglycan was higher in the cells of co-culture group [ (8.92±0.91) μg/L ] than in culture group [(2.56±0.26) μg/L,P ＜ 0.05], but lower than in the chondrocyte group[(13.69±1.51) μg/L, P＜ 0.05].CONCLUSION:Insulin-like growth factor-Ⅰ and chondrocyte culture solution can induce bone marrow-derived mesenchymal stem cells to differentiate into chondrocytes.

Objective To measure the expressions of programmed death?1(PD?1)and programmed death ligand?1(PD?L1)on peripheral blood T lymphocytes of patients with condyloma acuminatum(CA), and to investigate their role in cellular immunity in these patients. Methods Peripheral blood samples were obtained from 30 patients with CA(CA group)and 20 healthy human controls (control group). Flow cytometry was conducted to detect the expressions of PD?1 and PD?L1 on the surfaces of peripheral blood CD4+and CD8+T lymphocytes, and to determine the counts of CD4+and CD8+T lymphocytes. Enzyme?linked immunosorbent assay(ELISA)was performed to measure the levels of serum interleukin?2(IL?2)and interferon?γ(IFN?γ). Statistical analyses were carried out to compare the above parameters between the two groups, and to assess the relationship of PD?1 and PD?L1 expressions with the counts of CD4+and CD8+T lymphocytes as well as with the serum levels of IL?2 and IFN?γ. Results There was a significant increase in the expression rates of PD?1 and PD?L1 on CD4+T lymphocytes(PD?1:9.48%± 3.31%vs. 7.12%± 2.16%, t=2.81, P<0.01;PD?L1:4.40%± 1.46%vs. 3.26%± 1.13%, t=3.16, P<0.01)and CD8+T lymphocytes(PD?1:12.52%± 3.17%vs. 9.95%± 2.17%, t=3.16, P<0.01;PD?L1:7.07%± 2.23%vs. 5.39%± 1.69%, t=2.88, P<0.01)in the CA group compared with the control group. Moreover, the CA group showed significantly lower counts of CD4+T lymphocytes(727.43 ± 138.59/μl vs. 804.25 ± 92.83/μl, t=2.17, P<0.05)and CD4/CD8 ratio(1.23±0.35 vs. 1.46 ± 0.34, t = 2.24, P < 0.05) than the control group, while no significant difference was observed in CD8 + T lymphocyte counts between the CA group and control group(613.60 ± 121.60/μl vs. 572.45 ± 103.08/μl, t=1.24, P>0.05). The levels of serum IL?2 and IFN?γwere both lower in the CA group than in the control group(t=2.12, 2.16, respectively, both P < 0.05). In the CA group, PD?1 and PD?L1 expression levels on peripheral blood CD4 + T lymphocytes were both negatively correlated with CD4+T lymphocyte counts, the CD4/CD8 ratio, as well as IL?2 and IFN?γserum levels(all P<0.05), and those on peripheral blood CD8+T lymphocytes were also negatively correlated with the CD4/CD8 ratio(all P<0.05), but uncorrelated with CD8+T lymphocyte counts(both P>0.05). Conclusion PD?1 was highly expressed on peripheral blood T lymphocytes from patients with CA, which may inhibit T lymphocyte?mediated immune response, decrease CD4+T lymphocyte counts, the CD4/CD8 ratio as well as IL?2 and IFN?γserum levels by interacting with its ligand PD?L1 and forming the PD?1/PD?L1 signaling pathway.

BACKGROUND:Pain is the main clinical manifestation for ankylosing spondylitis. At present, nonsteroid anti-inflammatory drugs are oral y taken, but the effects are limited and toxic and side effects are more. Thus, there is no effective scheme for treatment of pain induced by ankylosing spondylitis.
OBJECTIVE:To investigate the correlation between postoperative joint pain al eviation and al ogeneic blood transfusion, and the mechanisms.
METHODS:We retrospectively analyzed clinical data of 88 ankylosing spondylitis patients combined with kyphosis who received only one section of osteotomy surgery merging hip joint pain. We compared the visual analog scale score of hip joint and detected the variation of leucocytes, lymphocytes and immunoglobulin concentrations before and after the operation in the groups of fresh al ogeneic whole blood transfusion, autologous whole blood transfusion, and mixed transfusion of al ogeneic and autologous whole blood. Flow cytometry was used to analyze the number and ratio of peripheral blood Th17 cells and Treg cells which were both highly associated with autoimmune diseases.
RESULTS AND CONCLUSION:The symptom of hip arthralgia obviously improved in both groups transfused by fresh al ogeneic whole blood or al ogeneic-autologous mixed whole blood. However, no obvious variation was detected in leucocytes, lymphocytes and immunoglobin concentration. However, flow cytometry results indicated that Th17/Treg proportion associated with autoimmune diseases was increased remarkably in peripheral blood of ankylosing spondylitis patients. Results suggested that al ogeneic whole blood transfusion can al eviate patients’ joint pain by correcting the imbalance of Th17/Treg which may improve their immune state.

Objective Glioblastoma,being one of the most malignant forms of adult brain tumors,is usually associated with a dismal prognosis.Given that the protein expression of O (6)-methylguanine-DNA methyltransferase (MGMT) is the most important determinant of temozolomide (TMZ) resistance,great efforts have been made to suppress it by regulating MGMT-related transcription factors.Resveratrol is a terpenoid that exhibits broad pro-apoptotic activity in various types of cancers,including glioblastoma.However,the effects of resveratrol on nuclear factor-κB (NF-κB) -MGMT signaling in glioblastomas have not yet been fully elucidated.In this article,we want to find that the molecular mechanisms of resveratrol reverses temozolomide resistance in glioblastoma cells.Methods Human malignant glioblastomas cell line T98G glioma cells was conventionally cultured in vitro; MTT assay was employed to detect the cell viability after being treated with dimethylsulfoxide (control group),or 100,200,400 and 800 μmol/L resveratrol,or 50,100,200 and 400 μmol/L TMZ or TMZ (100 μmol/L) combined with resveratrol (100 μmol/L); Hoechst33342 staining was employed to observe the effects of 100 μmol/L resveratrol,100 μmol/L TMZ or TMZ (100 μmol/L) combined with resveratrol (100 μmol/L) on T98G nuclear morphology changes; Flow cytometry and Western blotting were used to detect the T98G cell apoptosis and expressions ofMGMT,NF-κB signaling pathway related proteins.Results T98G cells after being treated with different concentrations of resveratrol for 24 and 72 h,the cell vitality decreased with 50％ inhibiting concentration (IC50) reaching 127.5 and 86.2 μmol/L,respectively; T98G cells after being treated with different concentrations of TMZ for 24 and 72 h,the cell vitality decreased with IC50 reaching 2.5 and 3.2 mmol/L,respectively; significant decreased cell vitality in the combination treatment with TMZ and resveratrol group was noted as compared with that in the TMZ or resveratrol treatment groups (P＜0.05).Hoechst 33258 staining and Western blotting revealed apoptotic morphological changes of T98G cell nuclei and increased apoptosis in the TMZ or resveratrol treatment groups,and more obvious changes were noted in the combination treatment with TMZ and resveratrol group.As compared with that in the control group,significantly increased MGMT protein expression in the 100 μmol/LTMZ treatment group was noted (P＜0.05),while no obvious MGMT protein expression was noted in the resveratrol treatment group and combination treatment with TMZ and resveratrol group; as compared with those in the control group,significantly increased IkB-α expression and decreased NF-κB p65 expression in the resveratrol treatment group (P＜0.05),significantly decreased IkB-α expression and increased NF-κB p65 expression in the TMZ treatment group (P＜0.05),and decreased NF-κB p65 expression in the combination treatment with TMZ and resveratrol group (P＜0.05).Conclusion Down-regulation of MGMT in T98G glioblastoma cells by NF-κB-dependent pathway is one of the important molecular mechanism of resveratrol reversing temozolomide resistance.