OBJECTIVETo explore the intervening and therapeutic effect of Cordyceps mycelia extract (CME) on liver cirrhosis induced by dimethylnitrosamine (DMN) in rats.

METHODSRat liver cirrhosis model was established by peritoneal injection of DMN at a dose of 10 microg/kg, once daily in the first 3 days of every week for 4 successive weeks. Experimental study on CME-intervention was conducted from the beginning of modeling to the end of the 4th week, while the CME-treatment experiment was carried out from the 4th week of modeling, when terminating the modeling factor, to the end of the 8th week, by administering CME at a dose of 0. 74 g/( kg d) once a day. Animals were killed in batches on the 3rd day, the 2nd (T1), 4th (T2), 6th (T3) and 8th (T4) week after modeling, to observe the histopathologic change in liver and the immunohistochemical staining of alpha-smooth muscle actin (alpha-SMA) and collagen type I (Col I), determine the content of hydroxyproline (Hyp) in liver, and the liver function was tested as well.

RESULTSCME-intervention experiment showed that as compared to those in the modeled rats at corresponding time points, in rats at T1 and T2, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity and total bilirubin (TBIL) content were significantly lower, and albumin (Alb) obviously higher; while at T2, Hyp content, ct-SMA and Col I positive expression were significantly lower (P < 0.05), the proliferation of collagen fibre attenuated. CME-treatment experiment showed that as compared to those in the modeled rats at corresponding time points, lower serum ALT, AST activity and TBIL content, and higher serum level of Alb were shown in rats at T1; and lower Hyp content, liver collagen fibre, and alpha-SMA positive expression were shown at T1 and T2; while less Col I positive expression at T2 was also shown in them (all P < 0.05).

CONCLUSIONCME could not only prevent the development of liver cirrhosis induced by DMN in rats, but also effectively promote the reversion of already formed liver cirrhosis, having a favourable prospect of clinical application.

Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Biological Factors ; therapeutic use ; Cordyceps ; chemistry ; Dimethylnitrosamine ; adverse effects ; Humans ; Liver ; metabolism ; Liver Cirrhosis ; chemically induced ; drug therapy ; enzymology ; genetics ; Male ; Moths ; chemistry ; Mycelium ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Treatment Outcome

OBJECTIVETo evaluate the role of serum level of procalcitonin (PCT), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and white blood cell count (WBC) as predictors in postoperative early infectious complications with fever after posterior lumbar internal fixation (PLIF).

METHODSA retrospective study was conducted from January 2012 to January 2014. Fifty-two patients with fever in the early stage(within 10 days) after the PLIF were collected in the study. They were divided into infection group and non-infection group (group A and group B) according to the results of postoperative blood culture. There were 26 patients in group A and 32 patients in group B. The values of PCT, CRP, ESR, and WBC count were compared and analyzed between two groups.

RESULTSThe values of PCT, CRP, and ESR in group A were higher than those of group B. Meanwhile, CRP and ESR in group B were still higher than the normal range. Among the 26 patients with infections (group A), PCT was superior to CRP and ESR, had a good ability in discriminating different kinds of postoperative infections. The area under the ROC curve of serum PCT levels was the largest (CI 95% was 0.81 to 0.98) in the indexs; and ROC curve of WBC count was no statistically significant. When the cut off points of each predictors were evaluated, the higher sensitive was CRP and reached at 90.27% and the higher specific was ESR and reached at 88.50%.

CONCLUSIONFor the patients with fever at the early stage after the PLIF should be paid attention and reasonable choosing predictors are helpful to identify postoperative infection in the early stage. The CRP and ESR may be influenced by the surgery, and the PCT level is helpful to differentiate infection type.

Adult ; Aged ; Aged, 80 and over ; Blood Sedimentation ; C-Reactive Protein ; analysis ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Female ; Fever ; blood ; diagnosis ; Fracture Fixation, Internal ; adverse effects ; Humans ; Infection ; blood ; diagnosis ; Leukocyte Count ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; blood ; diagnosis ; Protein Precursors ; blood

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. We have previously identified the benzoyl sulfathiazole derivative II as a non-competitive PTP1B inhibitor with in vivo insulin sensitizing effects. Preliminary SAR study on this compound series has been carried out herein, and thirteen new compounds have been designed and synthesized. Among them, compound 10 exhibited potent inhibition against human recombinant PTP1B with the IC50 value of 3.97 micromol x L(-1), and is comparable to that of compound II.
Humans ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Structure-Activity Relationship ; Sulfathiazoles ; chemistry ; pharmacology

Objective To develop a clinically useful assay for detecting the mutations of HBV pre-C/BCP based on the pyrosequencing and accuracy, reproducibility and reliability of this assay was evaluated. Methods The pyrosequencing primers for HBV pre-C/BCP mutation were designed through the cluster analysis among one hundred HBV gene sequences. After the amplification of the fragment of pre-C/BCP with the template of pre-C/BCP mutation plasmids, the pyrosequencing method for pre-C/BCP detection was initially set up with this standard sample. The accuracy, reliability and reproducibility of the pre-C/BCP pyrosequencing were confirmed through the pre-C/BCP plasmids as a standard sample when compared with Sanger/genechip sequencing method pre-C/BCP pyrosequencing assay was applied for detecting pre-C/BCP mutation types of 60 chnical serum samples in HBV patients. Results The pre-C/BCP mutation detection assay based on pyrosequencing has been established in our study. The coincidence rate between pyrosequencing and Sanger squencing was 100%. The coincidence rate between the result of pyrosequencing and of genechip method was 91.7%. The reproducibility of this assay was 97. 8%. It indicates the pre-C/BCP pyrosequencing is a high-accurate method with, good-reproducibility and high-reliability. And multi-site detection can be achieved by pyrosequencing one time. A rare mutation T1758C was also detected. Conclusion Pyrosequencing for pre-C/BCP mutations assay is high-throughout method for simultaneous detection of multi-site mutation.

BACKGROUND:With the deepening of bone tissue engineering research and bone metabolism understanding, it is a hotspot to analyze the blood supply and nutritional status of tissue-engineered bone.
OBJECTIVE:To compare different methods for evaluating smal vascular network distribution around the knee joint in rats in order to provide a guideline for the study of microvascular network in tissue-engineered bone.
METHODS:Eighteen Sprague-Dawley rats were randomly divided into three groups, with six rats in each group. Three commonly methods were used to evaluate the smal vascular network around the knee joint in rats:immunohistochemistry analysis, angiography analysis, and CT scans and reconstruction analysis.
RESULTS AND CONCLUSION:The microstructure of vascular network could be observed by immunohistochemistry, but the spatial distribution of vessels could not be evaluated. The spatial distribution of vessels could be showed by angiography and CT scans. However, some of micro vessels were showed unclearly by CT scans. The number of blood vessels detected by immunohistochemistry was (26.50±3.02) vessels, significantly higher than those detected by angiography and CT scans that were (14.12±1.47) and (9.00±1.79) vessels, respectively. Combination of immunohistochemistry and angiography can evaluate the microvascular network at microscopic and macroscopic levels, which can provide the whole information of the vascular network.

OBJECTIVETo observe if ER alpha gene can be induced by 5-aza-CdR in ER alpha negative human breast cancer cell lines (MDA-MB-231 and MDA-MB-435) and the synergistic inhibitory effects of 5-aza-CdR and Tamoxifen on these two cell lines in vitro.

METHODSThe status of 5'CpG island methylation of ER alpha gene in ER alpha negative (MDA-MB-231 and MDA-MB-435) human breast cancer cell lines and 20 cases of breast cancer tissue was studied by MSP, the expression of ER alpha mRNA was inspected by using RT-PCR after these two cell lines were treated with 5-aza-CdR. Cell proliferation was evaluated by MTT assay, distribution of cell cycle and rate of apoptosis were determined by flow cytometry after these two cell lines were treated with 5-aza-CdR or TAM alone, or in combination in vitro.

RESULTSThe 5'CpG island is methylated in the core promotor of ER alpha gene in ER alpha negative (MDA-MB-231 and MDA-MB-435) human breast cancer cell lines and the methylating rate is 25.0%, 66.7%, 83.3%, 100% in 20 cases of breast cancer tissue of stage I, II, III, IV, respectively. The expression of ER alpha mRNA was induced in these two cell lines after treated with 5-aza-CdR, MTT array showed the proliferation activity of these two cell lines was obviously reduced in 5-aza-CdR group and the inhibitory effect on proliferation was enhanced when 5-aza-CdR combined with TAM compared with control group, the induction of apoptosis was 11.20% and 8.71% respectively by 5-aza-CdR, while the rate of apoptosis is 48.8% and 53.1% when these two cells were treated with 5-aza-CdR combined with TAM.

CONCLUSIONS5-aza-CdR can re-express ER alpha by demethylating and sensitive ER alpha negative human breast cancer cell lines to TAM, 5-aza-CdR and TAM synergistically inhibit proliferation and induce apoptosis in ER alpha negative human breast cancer cell lines, thus offer a new way for the treatment of ER alpha negative breast cancer.

Adult ; Aged ; Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Azacitidine ; analogs & derivatives ; pharmacology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Drug Synergism ; Estrogen Receptor alpha ; biosynthesis ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Middle Aged ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tamoxifen ; pharmacology

OBJECTIVETo observe the effects of the external application of Qiyu oil gauze (QYOG) for promoting post-operational healing in patients with anal fistula and to explore its mechanism of action so as to provide a beneficial scientific basis for its wide use.

METHODSSixty patients with anal fistula scheduled to receive simple low anal fistulectomy were equally assigned, according to the sequence of hospitalization, to the tested group and the control group, and their wounds were classified according to longitudinal diameter into three grades (Grade I with a diameter below 2 cm; Grade II, 2-5 cm; and Grade III, over 5 cm). After the operation was completed and the operational wound was sterilized with benzalkonium bromide, the wound substratum was packed with QYOG in the test group and with vaseline gauze in the control group. The packing gauze was changed every day till the wound was healed. The healing time of the patients was observed, and the number of capillaries and positive cell percentages of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF) in wound granulation tissues were counted on the 5th day after the operation.

RESULTSThe wound healing time was 17.80+/-5.46 days in the test group, which was significantly shorter than that in the control group (21.90+/-6.32 days, P<0.01). The number of capillaries and positive cell percentages of VEGF and bFGF in wound granulation tissues on the 5th day in the tested group were higher than those in the control group (P<0.01), though the difference in EGF between the two groups was insignificant (P>0.05).

CONCLUSIONQYOG could shorten the wound healing time after anal fistulectomy, which suggests that it participates in the stimulation of wound granulation tissues to produce VEGF and bFGF, and thus promotes capillary genesis and improves blood circulation in wounds so as to promote wound healing.

Administration, Topical ; Adult ; Bandages ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Granulation Tissue ; drug effects ; Humans ; Male ; Middle Aged ; Plant Oils ; administration & dosage ; Postoperative Care ; methods ; Rectal Fistula ; drug therapy ; rehabilitation ; surgery ; Time Factors ; Treatment Outcome ; Wound Healing ; drug effects ; physiology

Adolescent ; Child ; Child, Preschool ; Clubfoot ; physiopathology ; Electromyography ; Female ; Humans ; Infant ; Male

OBJECTIVETo study the acting mechanism of Cordyceps mycelia extract (CME) for antagonizing hepatic sinusoidal capillarization (HSC) in rats with dimethylnitrosamine (DMN) induced liver cirrhosis.

METHODSRat liver cirrhosis model was established by peritoneal injection of DMN 10 mg/kg 3 times a week for 4 weeks. To rats in the CME-prevented group CME were administrated at a dose of 10 mL/kg, once a day, for 4 weeks. The observation time points were scheduled on the 3rd day (d3), and at the end of the 2nd (W2) and 4th week (W4) after modeling, and the following items were observed: hepatic ultrastructure was observed under electron microscope; expressions of CD44, von Willebrand factor (vWF) and type IV collagen (Col lV) in the liver sinusoidal walls by immunohistochemistry; matrix metalloproteinase-2 and-9 (MMP-2, MMP-9) activity under zymogram method; and serum hyaluronic acid (HA) content by radioimmunoassay.

RESULTSObservation at d3 showed MMP-2 and MMP-9 activity significantly increased, Col IV deposition and CD44 positive staining decreased, vWF positive staining increased in the liver sinusoidal walls, the fenestrae in the sinusoidal endothelial cells (SECs) decreased, and serum HA content increased (P<0.05); at W4, SECs defenestration and sub-SECs basal membrane formation were shown. In the CME-prevented group MMP-2 and MMP-9 activity significantly decreased (P<0.05); defenestration and basal membrane formation alleviated in the early stage (d3, W4); and at W2 and W4 decreases of HA content and vWF positive staining were shown, with increase of CD44 positive staining (P<0.05), more SECs fenestrae, and alleviated basal membrane formation.

CONCLUSIONSThe elevation of MMP-2 and MMP-9 activity in the early stage, which degrades the Col IV normally distributed under the sinusoidal endothelium, is an important factor for HSC formation. CME could inhibit the initiation of HSC by decreasing MMP-2 and MMP-9 activity in the early stage, and prevent its formation by decreasing SECs injury and phenotypic changes.

Animals ; Capillaries ; pathology ; Cordyceps ; Dimethylnitrosamine ; adverse effects ; Hepatic Veins ; cytology ; drug effects ; pathology ; Liver ; blood supply ; Liver Cirrhosis, Experimental ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mycelium ; Neovascularization, Pathologic ; prevention & control ; Rats ; Rats, Wistar

OBJECTIVETo evaluate the expression of telomerase transcriptional elements-interacting factor (TEIF) in human testis under different status and its relation with human telomerase reverse transcriptase (hTERT) expression.

METHODSSpecific antisera against TEIF were generated by immunization of rabbits with purified recombinated partial TEIF. Samples were assigned to three groups according to their pathological types, including 16 normal testes, 8 atrophic testes, and 6 testicular seminomas. They were subjected to immunohistochemical staining of TEIF and hTERT. Results from both TEIF and hTERT were analyzed semi-quantitatively and compared.

RESULTSThe expressions of TEIF and hTERT were detected in all samples of normal, atrophic testes, and seminomas. No differences of TEIF expressions among these three groups were observed (P > 0.05). On the contrary, the expressions of hTERT were significantly lower in atrophic testes compared with those of normal testes and seminomas (both P < 0.05). Nevertheless, co-expressions of TEIF with hTERT were revealed to be in normal and malignant cases (P < 0.05) but not in atrophic testes, which generally presented TEIF expression. The cellular distributions of both proteins were similar and mainly in spermatocytes and some Sertoli cells, while were all negative in the interstitial cells and other stromal cells. Conclusions The uniform expressions of TEIF in all these specimens suggest that it may be a marker of testis and its related diseases. The strong expression of hTERT in normal testes and testicular seminomas comparing with the low expression in atrophic testes may suggest a role for telomerase in maintaining proliferation of germ cells.

Atrophy ; Biomarkers ; DNA-Binding Proteins ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Seminoma ; metabolism ; Telomerase ; metabolism ; Testicular Neoplasms ; metabolism ; Testis ; metabolism ; pathology ; Transcription Factors ; metabolism