Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Bone Marrow ; metabolism ; Brain ; metabolism ; Brain Neoplasms ; metabolism ; pathology ; CDC2 Protein Kinase ; metabolism ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Glioma ; classification ; metabolism ; pathology ; Humans ; Male ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; Young Adult

OBJECTIVEOur previous cDNA array data have shown that expression level of CDK1 increased along with the malignant progression of ganglioglioma, and decreased with the differentiation process of neural stem cells. The purpose of this study was to investigate the CDK1 expression levels in gliomas and the effects of CDK1 knockdown on phenotype of glioma cells.

METHODSGlioma tissue array was constructed, which was composed of surgical specimens of gliomas with different malignancy grades, glioma xenografts in nude mice, cellular spheroids of brain tumor stem cells, normal neural stem cells and glioma cell line. CDK1 expression was detected in glioma tissue array with immunohistochemical techniques. CDK1 expression in human brain glioma cell line and relevant xenogeneic graft tumor was inhibited by retroviral vectors expressing short hairpin RNAs (shRNAs). Both in vitro and in vivo changes of biological characteristics were further observed.

RESULTSThe expression level of CDK1 increased along with the malignancy progression of glioma in clinical specimens. The positive expression rates of CDK1 in human brain glioma tissues were 22.2% (grade I), 40.0% (grade II), 69.6% (grade III) and 78.6% (grade IV), P = 0.01, respectively. The positive expression rate of CDK1 in glioma cell line and implanted xenografts was similar as the clinical tumors with high malignancy, and higher than those in neural stem cells and brain tumor stem cells (P = 0.0014). Expression of CDK1 was high in human fetal brain tissues and bone marrows of nude mice, but low in normal adult human brain tissues. Downregulation of CDK1 inhibited the proliferation activities notably both in SHG-44 cells in vitro and relevant xenogeneic graft tumors, and induced apoptosis of tumor cells prominantly as well.

CONCLUSIONOverexpression of CDK1 may promote oncogenesis and progression of human gliomas. Downregulation of CDK1 expression can inhibit the proliferation activities of human malignant gliomas.

Animals ; Apoptosis ; drug effects ; Astrocytoma ; genetics ; metabolism ; pathology ; Brain Neoplasms ; genetics ; metabolism ; pathology ; Brain Stem Neoplasms ; metabolism ; CDC2 Protein Kinase ; genetics ; metabolism ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Ganglioglioma ; genetics ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Mice ; Mice, Nude ; Neoplasm Staging ; Neoplasm Transplantation ; RNA, Messenger ; metabolism

BACKGROUNDNowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase II beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E. coli).

METHODSThe ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E. coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot.

RESULTSA 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase II beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E. coli JM109. The recombinant protein could be recognized by the S. japonicum infected rabbit serum.

CONCLUSIONThe full-length cDNA sequences encoding S. japonicum casein kinase II beta subunit were firstly sequenced, cloned, and expressed in E. coli.

Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Western ; Casein Kinase II ; chemistry ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; isolation & purification ; Escherichia coli ; genetics ; Molecular Sequence Data ; Rabbits ; Schistosoma japonicum ; enzymology ; genetics

OBJECTIVETo study the early change of serum nitric oxide (NO) after acute heat exposure with trauma and the effect of NO on mean arterial pressure (MAP), thus to provide theoretical basis for studying the mechanism of NO effect in acute stress.

METHODSThe rabbit model of acute heat exposure combined with trauma was established. The animals were divided into four groups, including control, trauma, hyperthermia and hyperthermia combined with trauma. The levels of NO were measured at different time points: 0 h, 1 h, 2 h and MAP was monitored throughout the whole experiment.

RESULTSThe concentration of NO declined at first and then increased at 1 h or so after acute heat exposure and trauma. The levels of NO in hyperthermia with trauma group at 1 h, 2 h were (42.75 +/- 8.24), (59.54 +/- 9.05) micro mol/L respectively (P < 0.05), while those in control group were (56.63 +/- 3.79) and (55.22 +/- 7.15) micro mol/L, the difference at 1h between two groups was significant (P < 0.05). Under the circumstance of hyperthermia and trauma, the level of MAP declined to the lowest point at 60 - 70 min and then showed a transient rise, after that, the level declined rapidly.

CONCLUSIONSAt the early stage of acute heat exposure and trauma, the concentration of serum NO declined at first and then increased, and had certain relationship with the change of MAP.

Animals ; Blood Pressure ; Cytokines ; biosynthesis ; Hot Temperature ; Male ; Nitric Oxide ; blood ; Rabbits ; Wounds and Injuries ; blood ; physiopathology

OBJECTIVETo find out the feasibility of tendon engineering in vitro using expanded tenocytes and polyglycolic acids (PGA).

METHODSTenocytes were isolated using tissue explant method and expanded in vitro. Tenocytes (20 x 10(6)) at the second passage were collected and then seeded onto PGA unwoven fibers to form a cell-scaffold construct in a shape of tendon. The constructs were cultured in DMEM with 20% FBS for 1 week. The cell-scaffold constructs were then cultured under constant tension generated by a U-shaped spring (n = 5), which served as experimental group, or cultured without tension (n = 4), which served as control group 1. PGA fibers alone were cultured (n = 3), which served as control group 2. Small fragments at the end of the constructs were harvested at 2, 4 and 6 weeks respectively for histological and immunohistochemistry (IHC) analysis. Six-week samples were also evaluated by transmission electron microscope (TEM) and mechanical test.

RESULTSNo obvious difference was observed among the three groups at 2 weeks grossly and histologically as the constructs remained to be mainly undegraded PGA fibers. By 4 weeks, a neo-tendon was formed in the experimental group and control group 1 grossly, and histology and IHC revealed the formation of collagen fibers. In contrast, PGA fibers alone in control group 2 were mostly degraded. At 6 weeks, tendons of control group 1 were much thicker [(2.55 +/- 0.18) mm in diameter] than those of experimental group [(1.44 +/- 0.13) mm in diameter]. Periodical striae were observed in collagen fibers of experimental group and control group 1 by TEM. However, histology of tendons in experimental group revealed longitudinally aliened collagen fibers, which resembled the structure of normal tendon more closely than that of control group 1 tendons. Furthermore, the maximum tensile stress (N/mm(2)) of experimental group (1.107 +/- 0.327) was greater than that of control group 1 (0.294 +/- 0.138) (P < 0.05).

CONCLUSIONIt is possible to use an engineering to construct tendon tissue in vitro. Periodical strain generated by bioreactor may be the optimal mechanical stimulation, which is currently under investigation.

Animals ; Cells, Cultured ; Polyglycolic Acid ; Tendons ; cytology ; growth & development ; Tissue Engineering ; methods

OBJECTIVETo determine the expression of SV40Tag, Rb and IRS-1 in gliomas and to identify their function in gliomagenesis and progression.

METHODSTissue microarrays were constructed containing 118 samples including human glioma and meningioma, experimental glioma, and normal human brain tissue. The expression of SV40Tag, Rb, IRS-1, SV40Tag combined with Rb, and SV40Tag combined with IRS-1 were assayed by immunofluorescence or immunohistochemical techniques. The expression ratio and level were analyzed.

RESULTSThe expressions of SV40Tag, Rb and IRS-1 were detected in gliomas and benign brain tumors. Their positive expression rate in glioma was 65.9%, 64.6% and 48.8%, respectively, with a statistically non-significant difference between the malignant and benign brain tumors. The malignant degree was positively correlated with SV40Tag and IRS-1, but negatively correlated with Rb expression. The combined expression rate of SV40Tag and Rb was 51.2%, and the combined expression rate of SV40Tag and IRS-1 was 40.2%. In the normal human brain tissue only the expression of Rb (77.8%, 7/9) and IRS-1 (22.2%, 2/9) were detected, but expression of SV40Tag could not be observed.

CONCLUSIONOur findings that no expression of SV40Tag was observed in normal human brain tissue indicates that expression of SV40Tag may play an important role in the pathogenesis of glioma. It may be assumed that after SV40 virus invading human body, Rb disfunction and IRS-1 activation promote the malignant transformation of cells, which could be one of important factors in pathogenesis and procession of glioms.

Adolescent ; Adult ; Aged ; Animals ; Antigens, Polyomavirus Transforming ; metabolism ; Brain ; metabolism ; pathology ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Neoplastic ; Glioma ; metabolism ; pathology ; Humans ; Insulin Receptor Substrate Proteins ; metabolism ; Male ; Meningioma ; metabolism ; pathology ; Mice ; Middle Aged ; Neoplasm Transplantation ; Rats ; Rats, Sprague-Dawley ; Retinoblastoma Protein ; metabolism ; Tissue Array Analysis ; Young Adult

OBJECTIVETo compare the enhanced recovery program after surgery (ERAS) with conventional perioperative management in patients undergoing radical resection for colorectal cancer.

METHODSThe ERAS protocol included a combination of evidence-based and consensus methodology. A total of 597 consecutive patients undergoing elective colorectal resection were randomized to either the ERAS(n=299) or the control group(n=298). Outcomes related to nutrition and metabolism index, stress index, and recovery index were measured and recorded.

RESULTSDemographics and operative parameters were similar between the two groups(P>0.05). The nutritional status of patients in the ERAS group was improved after surgery compared with that of the control group. On postoperative day (POD) 1, the HOMA-IR in the ERAS group was significantly lower than that in the control group(P<0.01). The cortisol level in the control group was elevated on both POD 1(P<0.01) and POD 5(P<0.01) compared to the preoperative level. However, the cortisol level was not increased until POD 5(P<0.01) in the ERAS group. The levels of TNF-α, IL-1β, IL-6, and IFN-γ were reduced in the ERAS group, indicating less postoperative stress responses compared with the control group. In addition, ERAS group was associated with accelerated recovery of gastrointestinal function. The postoperative length of stay [(5.7±1.6) d vs. (6.6±2.4) d, P<0.01] and expense[(15 998±2655) RMB vs. (17 763±3059) RMB, P<0.01] were reduced in the ERAS group. Twenty-eight patients(9.4%) in the control group and 29(9.7%) in the ERAS group developed complications, while the difference was not statistically significant(P>0.05).

CONCLUSIONERAS protocol alleviates surgical stress response and accelerates postoperative recovery without compromising patient safety.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Perioperative Care ; methods ; Prospective Studies ; Young Adult

OBJECTIVETo evaluate the efficacy of second allogeneic hematopoietic stem cell transplantation (allo-HSCT) for treatment of leukemia relapsed after first allo-HSCT.

METHODSNine patients with relapsed acute leukemia (5 AML, 4 ALL) and one with chronic myelogenous leukemia (CML) who showed cytogenetic relapse after first allo-HSCT received second allo-HSCT. The median relapse time from the first allo-HSCT was 141 days. Conditioning regimens for second allo-HSCT were combination chemotherapy based on moderate-dose Ara-C (n = 5), Bu (n = 3), conventional-dose Ara-C (n = 1) and Flud/Mel (n = 1). Prophylaxis for acute graft-versus-host disease (aGVHD) were CsA alone (n = 2), CsA/MTX (n = 1), FK506 (n = 1), and no prophylaxis in 6. The median number of peripheral blood mononuclear cells transfused was 6.1 x 10(8)/kg.

RESULTSEight cases were evaluable. All of them were engrafted and 7 developed aGVHD (grade I 4, grade II 3). The median time for absolute neutrophil count (ANC) > 0.5 x 10(9)/L and platelets > 20 x 10(9)/L were 11 and 12 days, respectively. Five cases developed localized chronic GVHD. Of all the 10 cases received second allo-HSCT, 8 died from interstitial pneumonia (n = 2), multiple-organ failure (n = 1), sepsis (n = 1), fungous pneumonia (n = 1), and leukemia relapse (n = 3), and 2 survived without leukemia for +986 and +1913 days, respectively. The leukemia free survival, transplantation related mortality and relapse rate at 2 year were 20%, 50% and 30%, respectively.

CONCLUSIONSecond allo-HSCT is a therapeutic alternative for selected patients with relapsed leukemia after first allo-HSCT.

Adult ; Disease-Free Survival ; Female ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; pathology ; surgery ; Male ; Neoplasm Recurrence, Local ; Retrospective Studies ; Transplantation Conditioning ; methods ; Transplantation, Homologous ; Treatment Outcome

Objective To observe the prevalence of hypertension and associated risk factors in the Guangxi Bai Ku Yao populations.Methods A toLal of 1170 subiects of Bai Ku Yao aged 15 and over were surveyed by a stratified randomized cluster sampling.Blood pressure,body height,weight,waist circumference,serum lipid and apolipoprotein levels were measured, and body mass index(BMI)were calculated,matched 1173 subjects of Han Chinese from the same region served as control.Results The standardized prevalence of hypertension in Bai Ku Yao was significantly lower than that in Han(11.53% vs.16.79%,P<0.01).The mean levels of systolic,diastolic blood pressure,and pulse pressure in Bai Ku Yao were also signifieanfly lower than those in Han[(115.7 ±16.3)vs.(120.0±16.3)mm Hg (1 mm Hg=0.133 kPa),P<0.01;(74.1 ±9.4)vs.(75.9 ±10.4)mm Hg,P<0.01;and(41.6 ± 12.0)vs.(44.2 ±11.2)mm Hg,P<0.01;respectively].Hypertension was positively correlated with male,age,physical activity,BMI,waist circumference,and the intakes of total energy,total fat,and sodium,and negatively associated with education level in both ethnic groups(P<0.05-0.01),but was positively associated with alcohol consumption only in Han.The rates of awareness,treatment and control of hypertension were significantly lower in Bai Ku Yao than those in Han population[(11.81%vs.21.76%),P<0.05;(5.51%vs.12.95%),P<0.05;and(2.36%vs.8.29%),P<0.05;respectively].Conclusion The prevalence of hypertension was significantly lower in Bai Ku Yao population than in Han population and diet,low sodium inmke,life style,and genetic factors might be responsible for the lower hypertension prevalence in Bai Ku Yao population.

OBJECTIVE: To study the clinical effect of individualized controllable stress external fixator in the treatment of open tibial fractures. METHODS: From December 2018 to July 2020, 60 patients with open tibial fractures were treated, including 35 males and 25 females;The age ranged from 23 to 58 years;The course of disease was 1.2 to 10.0 h. According to the stress stimulation on the fracture end after operation, all patients were divided into 4 groups, including non stress group (15 cases) and 3 groups with different stress stimulation(15 cases in each group). All patients with open tibial fractures were treated with controllable stress external fixator. Four weeks after operation, the stress group adjusted the elastic external fixator to apply axial stress of 1/6, 2/6 and 3/6 of their own weight to the fracture end based on the patient's weight. The wound healing of all patients after operation was observed, the plain CT images of fracture ends at 4, 6, 8, 10 and 12 weeks after operation were followed up, the average valueof callus area per 10 scanning planes was calculated, and the differences between the groups were compared. The fracture healing was observed and statistically analyzed. RESULTS: The wounds of all patients healed well, of which 7 patients underwent secondary free skin grafting and transferred myocutaneous flap. All patients were followed up for 12 to 24 months, with an average of 16.5 months. The final follow-up results showed that the fracture healing of stress groups and non stress group had significant difference( CONCLUSION When the controllable stress external fixation technique is used to treat open tibial fractures, the elastic external fixator is adjusted according to the patient's own weight after 4 weeks, and a certain axial stress is applied to the fracture end, which is conducive to the fracture healing of patients, and can reduce the incidence of delayed union or nonunion of open fractures, which has a certain application value.
Adult ; External Fixators ; Female ; Fracture Fixation ; Fracture Healing ; Fractures, Open/surgery* ; Humans ; Male ; Middle Aged ; Tibial Fractures/surgery* ; Treatment Outcome ; Young Adult