The Qinbai Qingfei concentrated pellets by traditional Chinese medicine theoryand party and group, the rats were given the drugs group, comparison of pharmacokinetics parameters changes of baicalin , discusses the rationality of Qinbai prescription. The rats were gavaged monarch drug group (Huang Qincu extract, mainly forbaicalin), and official medicine group, adjuvant group, medicine group and Qinbai group (Quan Fangzu) the content of baicalin equal as the monarch drug group, in the 28 h collection in rat plasma at different time point, application of HPLC determination of baicalin glycosides in rat plasmaconcentration time curve, with 3P97 practical pharmacokinetics program to process the data Based on the data analysis, baicalin in rat plasma of Qinbai group Cmax is 4 times as big as monarch druggroup, AUC is 6 times as big as monarch drug group; the content of baicalin in plasma of rats the highest is Qinbai group, the minister drug group, adjuvant group, medicine group of baicalin in rat plasma content of less than the Qinbai group, but was significantly higher than that of monarch drug group; the medicine group is slightly higher than that adjuvant the content of baicalin in plasma of rats. The pharmacokinetic results show that the measured plasma concentration in rats that Qinbai can significantly increase Cmax and AUC of baicalin, other components of qinbai can promoted the baicalin absorption in vivo. It showed that the reasonable of Qinbai compound compatibility. The minister drug can promote the absorption of baicalin in vivo.
Animals ; Area Under Curve ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Flavonoids ; blood ; pharmacokinetics ; Intestinal Absorption ; Male ; Rats ; Rats, Wistar

OBJECTIVETo construct T vectors based on Xcm I recognition site and optimize the PCR fragments for its ligation.

METHODSWe firstly cloned the human histone H4 cDNA containing one Xcm I recognition site at both its 5' and 3' end into pCDNA 3.0 vector and then generated T vector with pCDNA 3.0 backbone by cutting the recombinant plasmid with Xcm I. To increase the ligation efficiency, the primers were firstly phosphorylated before DNA fragments amplification and then the PCR products were treated with Taq DNA polymerase and dATP after PCR amplification. Two DNA fragments with the length of 312 bp and 1 329 bp were ligated to it and the ligation mixture was transformed into E. coli DH5α competent cells and the positive rates of the transformants were evaluated by PCR and DNA agarose gel electrophoresis.

RESULTSOur results showed that the T vector produced by our method could ligate to the target DNA fragments with high efficiency. Besides, the phosphorylation state of the primers used for PCR amplification is also an important factor determining the cloning efficiency. What's more, as for longer DNA fragments, retreatment with Taq DNA polymerase and dATP after PCR amplification and purification could improve the ligation efficiency significantly.

CONCLUSIONOur protocol may overcome the dependence on blue/white screening to get positive clones and provide a potent way to generate T vectors and ligate them to the target PCR fragment.

Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Histones ; genetics ; Humans ; Polymerase Chain Reaction ; methods

OBJECTIVETo summarize the experience and short-term clinical outcomes of hand-assisted laparoscopic surgery (HALS) in sphincter-preserving surgery for low and ultralow rectal cancer.

METHODSData of 49 patients with rectal cancer who underwent HALS for low or ultralow anterior resection between January 2010 and January 2011 were analyzed retrospectively.

RESULTSThe proximal resection margin was (14.3±6.9) cm and the distal margin was(4.3±1.9) cm. The mean operative time was(128.3±70.9) min. On postoperative macroscopic evaluation, the mesorectum was intact in 42 cases, nearly intact in 7 cases. The circumferential resection margin was more than 2 mm in 42 cases, and less than 2 mm in 7 cases. Forty-six patients underwent R0 resection, and 3 cases underwent R1 resection. The median retrieved lymph node (LN) was 16.20±9.23, and the median positive LN was 1.12±2.19. Postoperative pathological examination showed TNM stage was I( in 12 patients, II(A in 18, II(B in 1, III(A in 2, III(B in 8, III(C in 5, IIII( in 3. The median postoperative hospital stay was (6.25±3.87) d. There were no anastomotic leakage, ileus, intra-abdominal or anastomotic bleeding. There were two wound infections.

CONCLUSIONLow and ultralow anterior resection for rectal cancer using HALS approach is safe and feasible with favorable short-term outcome.

Adult ; Aged ; Aged, 80 and over ; Anal Canal ; surgery ; Female ; Hand-Assisted Laparoscopy ; methods ; Humans ; Male ; Middle Aged ; Rectal Neoplasms ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVET o summarize the clinical effects of the repairing methods for skin and soft tissue defection of heel.

METHODSFrom June 1998 to June 2009,42 patients with skin and soft tissue defection of heel underwent the repairing treatment,including 23 males and 19 females, with an average age of 37 years old ranging from 18 to 65. The causes of injuries included mangled injury in 22 cases, high fall injury in 10 cases, cut injury in 5 cases,melanoma in 3 cases, decubital ulcer in 2 cases. Of the 42 cases, 27 were on left side and 15 on right side. The defect area of skin ranged from 3 cm x 2 cm to 18 cm x 16 cm. The time between the injury and surgery ranged from 8 hours to 10 years. The wounds were repaired separately by medial plantar flap in 13 cases, lesser saphenous sural nerve vascular island flap in 18 cases, saphenous neurocutaneous vascular flap in 11 cases. The patients' outcome were evaluated with appearance,blood supply, texture, resilience and two points discrimination of the flaps.

RESULTSAll of the 42 flaps were survived. The distal skin necrosis occurred in 2 flaps, but healing occurred after debridement and intermediate thickness skin grafting. Three patients with sinus formation healed after 5 to 12 months of dressing change. All patients were follow-up for 8 months to 6 years. The flaps of all patients gained a satisfied shape after operation. The patients had a normal gait, the flaps had a good sense and a resistance to wearing,and no ulcer occurred. The two point discrimination of the flap was 4 to 12 mm.

CONCLUSIONIt is convenient and effective to repair the heel skin and soft tissue defects using medial plantar island skin flap when the defects is less then 8 cmx6 cm. As reliable blood supply,major artery preservation and high survival, the lesser saphenous sural nerve vascular island flap and saphenous neurocutaneous vascular flap can be transferred to repair the large soft tissue defect of heel.

Adolescent ; Adult ; Aged ; Female ; Foot Injuries ; surgery ; Heel ; abnormalities ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; Young Adult

Ligustrazine, one of the major effective components of the Chinese traditional medicinal herb Ligusticum Chuanxiong Hort, has been reported plenty of biological activities, such as protect cardiovascular and cerebrovascular, neuroprotection and anti-tumor, et al. Because of its remarkable effects, studies on structural modification of ligustrazine have attracted much attention. Ligustrazine synthetic derivatives reported in recent decades are mainly derived from four primary intermediates (TMP-COOH, TMP-OH, TMP-NH2, HO-TMP-OH). To explore the neuroprotection activitiy of ligustrazine intermediates, six ligustrazine intermediates (2, 5, 8, 11, 12, 13) were synthesized and their protective effects against CoCl2-induced neurotoxicity in differentiated PC12 cells were studied. The target compounds were prepared via different chemical methods, including oxidation, substitution, esterification and amidation without changing the structure nucleus of ligustrazine. Compared with TMP (EC50 = 56.03 micromol x L(-1)), four compounds (2, 5, 12 and 13) exhibited higher activity (EC50 < 50 micromol x L(-1)) respectively, of which, compound 2 displayed the highest protective effect against the damaged PC12 cells (EC50 = 32.86 micromol x L(-1)), but target compounds 8 and 11 appeared lower activity (EC50 > 70 micromol x L(-1)). By structure-activity relationships analysis, the introduction of carboxyl, amino to the side chain of ligustrazine and appropriately increase the proportion of ligustrazine may contribute to enhance its neuroprotective activity, which provides a reference for the design, synthesis and activity screening of relevant series of ligustrazine derivatives in the future.
Animals ; Cell Differentiation ; drug effects ; Chemistry Techniques, Synthetic ; Cobalt ; toxicity ; Drugs, Chinese Herbal ; chemistry ; Neuroprotective Agents ; chemical synthesis ; chemistry ; pharmacology ; Neurotoxins ; toxicity ; PC12 Cells ; Pyrazines ; chemical synthesis ; chemistry ; pharmacology ; Rats

OBJECTIVETo develop a method for ng quantitation of circulating DNA in serum and explore the value in the diagnosis of cancer.

METHODSSerum DNA was extracted by commercial "genomic DNA extraction kit" and detected by fluorescent dye (SYBR green I) staining. Loss of heterozygosity (LOH) at BRCA1 (D17S579, D17S855) and p53 (TP53, D17S786) in serum DNA was analyzed by PCR-based method.

RESULTSSYBR green I dot staining could detect DNA as low as 2 ng. Using this method, we detected serum samples from 483 patients with various types of cancer and 150 healthy individuals. The mean DNA concentration in the normal controls was 22.2 +/- 13.4 ng/ml, while that in cancer patients was 81.3 +/- 98.3 ng/ml (P < 0.001). In 33 ovarian cancer patients with increased DNA level, 27(81.8%) displayed LOH in at least one of the four loci analyzed.

CONCLUSIONCirculating DNA in serum may become additional tumor marker for the diagnosis of cancer.

BRCA1 Protein ; genetics ; Biomarkers, Tumor ; blood ; DNA, Neoplasm ; blood ; genetics ; Female ; Genes, BRCA1 ; Genes, p53 ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Neoplasms ; blood ; genetics ; Tumor Suppressor Protein p53 ; genetics

BACKGROUNDSeveral candidate genes of ossification of the posterior longitudinal ligament (OPLL) susceptibility have been identified, but their polymorphisms account for only a small percent of the total variance. Bone morphogenetic protein-4 (BMP4) is a potent ectopic ossification inducing factor. BMP4 protein and mRNA are present in cells from OPLL patients, but not non-OPLL controls. A single nucleotide polymorphism of 6007C>T(rs17563) of BMP4 has been reported to affect bone density in postmenopausal women. Thus, BMP4 may function in OPLL development. Appropriately, the relationship between BMP4 polymorphisms and OPLL was investigated.

METHODSA case-control association study investigated the genetic etiology in 179 OPLL patients and 298 non-OPLL controls. Extent of OPLL was analyzed by radiologic examinations. Whether single nucleotide polymorphism (SNP) of -5826G>A(rs1957860) 5' of the transcription start site and 6007C>T(rs17563) in exon 4 of the BMP4 gene were statistically associated with genetic susceptibility to OPLL in Chinese Han subjects was assessed.

RESULTSA significant statistical difference in genotype of 6007C>T polymorphism between male OPLL patients and male controls was evident, and the frequency of "TT" genotype in male OPLL patients was significantly higher than in male controls (P = 0.039). The frequency of the "T" allele was also significantly higher in male OPLL subjects than in male controls (P = 0.014, OR = 1.57). A significant difference was also observed between the 6007C>T polymorphism and the number of ossified cervical vertebrae in OPLL patients, while no statistical difference was apparent between the -5826G>A polymorphism and OPLL occurrence.

CONCLUSIONSThe T allele in the 6007C>T polymorphism may be a risk factor for male Han Chinese with ossification of the posterior longitudinal ligament in the cervical spine. Chinese Han male patients with CT and TT 6007C>T genotypes have a genetic susceptibility to OPLL and more extensive OPLL in the cervical spine.

Alleles ; Asian Continental Ancestry Group ; genetics ; Bone Morphogenetic Protein 4 ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; genetics ; Polymorphism, Single Nucleotide ; genetics

Objective To discuss the clinical curative effect of percutaneous vertebroplasty(PVP)combined with percutaneous pedicle screw fixation for thoracolumbar fracture.Methods Retrospectively analyzed the clinical data of 43 patients with thoracolumbar fracture who underwent PVP combined with percutaneous pedicle screw fixation in our hospital from November 2015 to June 2017.Those patients included 28 males and 15 females,and the age of patients ranged from 50 to 66 years old,with an average age of(58.26 ±3.67)years old.The func-tional outcome were evaluated by VAS scores and ODI scores before and after the operation.The sagittal Cobb angle was used to evaluate the reduction of fracture.Results All these patients all successfully completed the operation,and there was no complications after operation.The operation time ranged from 60 to 126 min,with an average time of(96.07 ±15.69)min;the blood loss ranged from 60 to 180 min,with an average time of(113.26 ±24.7)min.All the patients were followed up for 4 to 23 months,with an average time of(12.07 ±4.01)months. The VAS score,ODI score and sagittal Cobb angle were significantly decreased in the last follow -up period compared with those before surgery,and the difference was statistically significant(P<0.05).Conclusion PVP combined with percutaneous pedicle screw fixation in the treatment of thoracolumbar fracture has smaller incision,less blood loss,shorter operation time and better improvement of local pain,func-tional movement and kyphosis.

OBJECTIVETo investigate COX-2 expression in patients with gastric cancer and its relationship with angiogenesis and clinicopathologic features of gastric cancer.

METHODSCOX-2 expression and CD34-stained microvessel density (MVD) were detected by immunohistochemical methods in specimens from 96 patients with gastric cancer. The correlations among COX-2 expression, MVD and clinicopathologic features were analyzed.

RESULTSThe COX-2 positive rate and MVD in gastric cancer were significantly higher than those in the normal gastric mucosa (80.2% vs. 13.3%; 32.5+/- 8.3 vs. 13.1+/- 2.4, all P< 0.01). The COX-2 positive rate and MVD in the patients with stage III and IV were significantly higher (91.4% and 34.9+/- 8.7 respectively, P< 0.01), than that in the patients with stage I and II. The COX-2 positive rate and MVD in the cases with lymph node metastasis were 87.9% and (35.0+/- 8.5) respectively, higher than those in the cases without lymph node metastasis (P< 0.05). The Spearman rank correlation test showed a significant correlation between COX-2 expression and tumor MVD (r=0.311, P< 0.01).

CONCLUSIONSCOX-2 plays an important role in gastric cancer angiogenesis. COX-2 and angiogenesis induced by COX-2 contribute to tumor invasion and lymph node metastasis.

Adult ; Aged ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; metabolism ; pathology ; Stomach Neoplasms ; metabolism ; pathology

BACKGROUND: Stent-assisted coil embolization has become an important technique for the treatment of intracranial complex aneurysms. However, there is no systematic analysis of complications and prevention strategies related to this technique. OBJECTIVE: To systematically analyze the complications related to stent-assisted coil embolization of intracranial aneurysms during perioperative and follow-up periods and to assess the safety of this method. METHODS: A retrospective study of 232 consecutive patients with 239 wide-necked aneurysms who underwent stent-assisted coil embolization at the Department of Neurosurgery, General Hospital of Shenyang Military Command from July 2003 to December 2009 was performed. Angiographic results and clinical outcomes were evaluated. A variety of complications were analyzed. The technical feasibility of the procedure, procedure-related complications, angiographic results, clinical outcomes and follow-up angiography were evaluated. RESULTS AND CONCLUSION: Stenting was successful in the 236 of 239 aneurysms. Procedure-related complications included thomboembolism (n=13), intraprocedural rupture (n=8), coil protrusions (n=5), new mass effect (n=3), vessel injury (n=3), and stent dislodgement (n=2). Procedure-related morbidity and mortality was 4.3% (10/232) and 1.3% (3/232), respectively. Nonprocedural complications attributable to subarachnoid hemorrhage in 129 patients with acute ruptured aneurysms were symptomatic vasospasm (18.6%, 24/129) and shunt-dependent hydrocephalus (7.0%, 9/129). Favorable clinical outcomes (mRS 0-2) were observed in 88.3% (205/232) of the patients, and no rehemorrhage of treated aneurysms occurred. Follow-up angiography was obtained in 155 patients (159 treated aneurysms). The overall recanalization rate was 14.5% (23/159). No stenting or coiling displacement occurred during the follow-up. Delayed complications included in-stent stenosis (n=2) and penetrating artery occlusion (n=2). In conclusion, our study indicates that stent-assisted coil embolization of intracranial aneurysm is a safe technique with low morbidity and mortality rates. Thromboembolism is the main cause of procedure-related disability and death. Nevertheless, further long-term follow-up is necessary to determine the durability of these promising results.