ObjectiveTo explore clinical effect and safety ofShaoyao-Gancao decoction combined withSiwu decoction for the patients with diabetic peripheral neuropathy (DPN).MethodsA total of 76 patients with DPN in our hospital from 2013 July to October 2015 were divided into control group and observation group by the random number method, 38 patients in each group. The control group were treated with conventional treatment combined with oryzanol injection, and the observation group were treated with Shaoyao-Gancaodecoction combined withSiwu decoction on the basis of control group. Both groups were treated for one month. The level of FPG, 2 hPG, HbAlc, TC, TG, LDL-C and HDL-C was detected respectively before and after treatment. The whole blood viscosity and plasma viscosity was detected with LBY-N6A self-cleaning rotary viscosimeter. The red blood cells deposited and platelet aggregation rate were tested with Winchester points vascular method and turbidimetric method, respectively.The leg nerve and sural nerve of nerve conduction velocity (nerve conduction velocity, NCV) were determined with electromyography, at the same time the clinical curative effect was evaluted.ResultsTotal effect rate of observation group was 89.5% (34/38), which was significantly higher than the control group 71.1% (27/38), and the difference was statistically significant (χ2=4.070,P=0.044); After treatment, the levels of FPG (6.12 ± 0.38 mmol/Lvs.6.58 ± 0.52 mmol/L, t=4.403), 2 hPG (7.83 ± 0.82 mmol/Lvs. 8.41 ± 0.93 mmol/L,t=2.884), HbAlc (6.27 ± 0.52%vs. 6.82 ± 0.64%, t=4.112) in the observation group were lower than those in the control group (P<0.05). The levels of TC (4.73 ± 0.83 mmol/L vs. 5.11 ± 0.64 mmol/L,t=52.235), TG (1.83 ± 0.35 mmol/Lvs. 2.03 ± 0.41 mmol/L, t=2.287) and LDL-C (2.91 ± 0.54 mmol/Lvs. 3.25 ± 0.58 mmol/L,t=2.645) in the observation group were lower than those in the control group (P<0.05), and the HDL-C (1.47 ± 0.33 mmol/Lvs. 1.26 ± 0.31 mmol/L,t=-2.859) in the observation group was higher than than that in the control group(P<0.05). The level of whole blood specific viscosity (10.16 ± 2.12 mPa?svs. 11.33 ± 2.51 mPa?s,t=2.195), plasma viscosity (1.24 ± 0.25 mPa?svs. 1.62 ± 0.37 mPa?s,t=5.246), red blood cell volume (0.31 ± 0.16L/Lvs. 0.42 ± 0.08 L/L,t=3.791) and platelet aggregation rate (50.21% ± 8.03%vs. 54.16% ± 7.82%,t=2.172) in the observation group were significantly lower than those in control group (P<0.05). The tibial nerve velocity (45.22 ± 3.20 m/svs. 38.26 ± 5.19 m/s,t=-7.037) and the common nerve velocity (43.22 ± 6.34 m/svs. 36.23 ± 4.81,t=-5.415) in the observation group were as significantly higher than those in control group (P<0.05).ConclusionsThe Shaoyao-Gancaodecoction combined withSiwu decoction could improve the state of blood sugar, blood lipid and blood rheology of patients with DPN, and improve nerve transfer speed of leg nerve and sural nerve, the curative effect is better than conventional western medicine therapy alone.

Objective To explore effect of fetal lymphocyte on pathogenesis of intrahepatic cholestasis of pregnancy(ICP).Methods Twenty pregnant women with ICP and 20 normal pregnant women were enrolled in the study.The single mixed lymphocyte culture/reaction(MLC/MLR)was conducted using inactive lymphocyte obtained from maternal peripheral blood and lymphocyte of cord blood from fetus.Antigen-induced-lymphocyte-proliferation-reaction was used for dermic soluble antigen and decidual soluble antigen obtained from maternal blood and cord blood from fetus.The intense of proliferation was calculated and compared between normal and ICP-complicated pregnancies.Results(1)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 2.75?0.36 than those of normal control group 1.45?0.19 in single mixed lymphocyte culture(P＜0.05).(2)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group 1.45?0.19 than those of normal control group 0.67?0.24 in decidual soluble antigen induced lymphocyte proliferation reaction(P＜0.05). (3)The level of intense of proliferation of fetal lymphocyte was significantly increased in ICP group(1.22?0.44)than those of normal control group(0.66?0.27)in dermic soluble antigen induced lymphocyte proliferation reaction.Conclusions(1)The fetal lymphocyte may be one of the effector cells in pathogenesis of ICP.(2)The disturbance of fatal-maternal immune-tolerance is one of the important mechanisms underlying ICP.

OBJECTIVETo investigate the expression profiles of serum Golgi protein-73 (GP73) in liver cirrhosis and primary hepatic carcinoma (PHC) and determine its clinical value for differential diagnosis.

METHODSSerum protein expressions of GP73 and alpha-fetoprotein (AFP) were detected by enzyme-linked immunosorbent assay and chemiluminescence assay, respectively, in patients with PHC (n=80), liver cirrhosis (n=65), and healthy controls (n=50). Inter-group changes were assessed by Kruskal-Wallis test, and significance of these differences was assessed by Mann-Whitney test. A receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency and determine the cut-off values for GP73 and AFP. Sensitivity and specificity were compared by the Chi-squared test. Correlation between serum GP73 expression and clinical parameters was determined by Spearman's rank correlation analysis.

RESULTSThe PHC group showed significantly higher serum GP73 (282.0 mug/L) than the liver cirrhosis group (211.8 mug/L) and control group (58.3 mug/L) (H = 93.30, P less than 0.01). For differential diagnosis of PHC and liver cirrhosis, the cut-off value was 318.1 mug/L for GP73 and 13.4 mug/L for AFP. Sensitivity of GP73 was lower than AFP (45% (36/80) vs. 65% (52/80); X2 = 8.02, P less than 0.05). Specificity of GP73 was lower than AFP but no significance was found (83.1% (54/65) vs. 87.7% (57/65); X2=0.27, P more than 0.05). The areas under the ROC curves were not significantly different between GP73 and AFP (0.65 (95% confidence interval (CI): 0.54~0.72) vs. 0.75 (95% CI: 0.67~0.83); Z = 1.88, P more than 0.05). The area under the ROC curves increased but not significantly (0.80 (95% CI: 0.73~0.88) vs. 0.75 (95% CI: 0.67~0.83); Z=2.61, P more than 0.05). Serum GP73 was correlated with liver cirrhosis (r=0.27), vascular invasion (r=0.29), and TNM staging (r=0.27) (all P less than 0.05), but not with sex (r=0.13), age (r=0.10), enhanced AFP (> 13.4 mug/L; r=0.03), tumor size (r=0.18), or distant metastasis (r=0.04), all P less than 0.05.

CONCLUSIONSerum GP73 and AFP have comparable diagnostic efficiency, but the sensitivity of AFP is superior for differential diagnosis of liver cirrhosis and primary hepatic carcinoma. Elevated serum GP73 may be correlated with liver tumor load and aggressiveness.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; Case-Control Studies ; Diagnosis, Differential ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; Liver Neoplasms ; diagnosis ; Male ; Membrane Proteins ; blood ; Middle Aged ; Sensitivity and Specificity ; Transcriptome ; alpha-Fetoproteins ; metabolism

The growth characteristics and ginsenosides isolation of the suspension-cultured crown gall of Panax quinquefolium were studied. The result showed that the maximum biomass of cultures was 18.6 g/L (dry weight) and the content of ginsenosides reached its maximum level of 620.4 mg/L on the 27th day. The utilization rates of sugar, phosphorus, nitrogen in NH4+ and nitrogen in NO3- were 91.8%, 100%, 81% and 97%, respectively. Four compounds were isolated from the suspension-cultured crown gall and their structures were elucidated as pseudoginsenoside F11 (I), ginsenoside Rd (II), ginsenoside Rb1 (llI) and ginsenoside Rb3 (IV).
Culture Media ; Culture Techniques ; methods ; Ginsenosides ; biosynthesis ; isolation & purification ; Panax ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism

AIMTo create a simple behavioral procedure for detecting tinnitus of rats.

METHODSA procedure based on a Pavlovian-conditioned suppression paradigm was set referencing Jastreboff's way to detect tinnitus behaviorally in conscious animals. Rats received acquisition training through Pavlovian conditioning to associate the conditioned stimulus (noise offset) with the unavoidable unconditioned stimulus (foot-shock). The manifestation here is escape reaction. An extinction procedure followed, during which the subject was exposed to the conditioned stimulus, but shock is no longer given. The manifestations of the rats in different group during the extinction processes were observed. Tinnitus was induced by salicylate (SA) injection to validate the method.

RESULTSThe escaped rate of SA group is lower than that of the control. It indicated that the SA rats escaped seldom when there was a sound off, because the rats might perceive the salicylate-induced auditory sensation as a substitute for the background noise that had acquired a safety value during training.

CONCLUSIONThe behavioral procedure could be used to detect tinnitus of rats induced by salicylate.

Animals ; Conditioning (Psychology) ; Disease Models, Animal ; Escape Reaction ; Male ; Psychometrics ; methods ; Rats ; Rats, Wistar ; Tinnitus ; diagnosis

OBJECTIVETo investigate the status of male reproductive health among middle-aged and older men in the urban area of Nanjing.

METHODSWe collected the laboratory results of 884 middle-aged and older men aged 55 - 89 years from the Xuanwu District of Nanjing present for routine physical examinations, including those of blood routine tests, liver and kidney function, blood glucose, blood lipid, and total prostate specific antigen (TPSA), as well as such reproductive hormone indexes as total serum testosterone (TT), free serum testosterone (fT), and sex hormone binding globulin (SHBG). We also obtained the above reproductive hormone indexes from 119 young and middle-aged men aged 20 - 39 years as controls.

RESULTSAging-related changes were found in the 50 percentiles of all the reproductive hormones and relevant parameters but those of TT and E2, with gradual increases in LH, FSH and SHBG and decreases in fT, TSI and fTI. Comparison of reproductive hormones and relevant parameters by Mann-Whitney U test did not show any statistically significant differences in the TT level between any two of the five age groups (20 - 39, 55 - 59, 60 - 69, 70 - 79, and > or = 80 yr) (P > 0.05) except between the control and > or = 80 yr groups and the 60 - 69 and > or = 80 yr groups (P < 0.05), nor in the E2 level between any two groups, nor in the levels of LH and FSH except between the 55 - 59 and 60 - 69 yr groups and the 70 - 79 and > or = 80 yr groups, and nor in the levels of fT and TSI except between the 55 - 59 and 60 - 69 yr groups. However, there were significant differences in the levels of SHBG and fTI between any two age groups. Spearman correlation analysis revealed that fT, TSI, and fTI were correlated negatively with aging and LH (P < 0.05, I r I > 0.5) but weakly positively with cholesterol, blood glucose and hemoglobin (P < 0.05, /r/ < 0.5), SHBG and LH positively with aging, SHBG weakly negatively with blood glucose and hemoglobin, LH weakly negatively with hemoglobin, and TT weakly negatively with aging but positively with hemoglobin.

CONCLUSIONThe levels of serum testosterone, particularly that of fT, declined with aging in middle-aged and older men in the urban area of Nanjing, which may contribute to abnormal lipid metabolism, low hemoglobin and high blood glucose.

Aged ; Aged, 80 and over ; Aging ; blood ; Blood Glucose ; analysis ; Case-Control Studies ; China ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Sex Hormone-Binding Globulin ; analysis ; Statistics, Nonparametric ; Testosterone ; blood

OBJECTIVETo study the anti-tumor effects of all-trans retinoic acid (ATRA) and mechanisms of its action.

METHODSHuman esophageal carcinoma cell line EC9706 cells were treated with ATRA at different concentration. The proliferation inhibition was examined by MTT assay. Morphological examination, TUNEL method and flow cytometry were used to detect the apoptosis and changes of cell cycle. Immunohistochemical method was used to detect the expression of apoptosis-related genes caspase-3 and bcl-2. The semi-quantification of protein expression was analyzed by pathological image analysis.

RESULTSATRA inhibited the proliferation of EC9706 cells moderately. Apoptosis in EC9706 cells was induced by ATRA treatment. The morphology of EC9706 cells showed changes such as nuclear chromatin condensation and fragmentation. Sub-G1 peak was found by flow cytometry. The maximal apoptosis rate was 32.6%. The expression of caspase-3 gene was enhanced. The expression of bcl-2 gene was decreased. All these effects were presented in a dose-dependent and time-depend manner.

CONCLUSIONApoptosis is one of the key mechanisms of ATRA action on EC9706 cells.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tretinoin ; administration & dosage ; pharmacology

OBJECTIVETo evaluate the advantages, disadvantages and their indications of total nasal reconstruction with different techniques.

METHODSA series of total nasal reconstruction were treated with four methods from 1975 to 2003. These methods were tubed flap of arm,midline forehead flap with skin graft, midline forehead flap with bilateral frontotemporal flaps for repairing the donor site, and expanded forehead flap.

RESULTSAll of the patients were treated successfully. The shape and function of the reconstructed noses were satisfactory. However, the traditional forehead flap with skin graft may leave a unsightly big and black scar on the forehead. The technique of the tubed flap of arm could provide enough tissue without remaining forehead scar and be easily shaped, but it required long period, multiple procedures and body fixation for three weeks.

CONCLUSIONSMidline forehead flap with bilateral frontotemporal flaps for repairing the donor site may be good for small nose reconstruction while expanded forehead flap could reconstruct a big nose. Tubed flap of arm may be used to the patients who do not wish to leave any scar on the forehead. Forehead flap with skin graft to repair the donor sit- should generally be avoided for nose reconstruction.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Nose ; surgery ; transplantation ; Nose Deformities, Acquired ; surgery ; Reconstructive Surgical Procedures ; methods ; Reproducibility of Results ; Retrospective Studies ; Skin Transplantation ; methods ; Surgical Flaps ; Treatment Outcome

OBJECTIVETo investigate the frequency of microsatellite instability (MSI) and loss of heterozygosity (LOH) at human leucocyte antigen(HLA) class I loci in cervical carcinoma and the detailed deletion mapping in this region.

METHODSLOH and MSI of HLA class I genes were analyzed in 30 paired blood and tumor samples by PCR based single-stranded length polymorphism (PCR-SSLP).

RESULTSOf the 30 cases, 23(76.7%) showed LOH at one or more loci. Higher frequencies of LOH were found at four loci: C3_2_11 (50%), C1_4_4 (37%), C1_2_5 (36.7%), D6S276 (48.3%). MSI was found in 20 out of 30 cases (66.7%).

CONCLUSIONThe data suggest that the LOH and MSI of HLA class I gene might participate in the carcinogenesis of cervical carcinoma. Meanwhile, the minimal deletion region might be defined between C1_2_5 and C3_2_11, thus providing the evidence for cloning the tumor suppressor genes associated with cervical carcinoma.

Adult ; Aged ; Female ; Histocompatibility Antigens Class I ; genetics ; Humans ; Loss of Heterozygosity ; genetics ; Microsatellite Instability ; Middle Aged ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; genetics

OBJECTIVETo construct an eukaryotic expression vector of human stathmin gene and to assess its effect on esophageal cancer EC9706 cells.

METHODSStathmin cDNA coding sequence was amplified by RT-PCR from Eca109 cells and was cloned into pMD18-T vector. After identifying and sequencing, the correct inserting stathmin gene was sub-cloned into eukaryotic expression vector pEGFP-C2. EC9706 cells were transfected with this recombinant plasmid and control plasmid using Lipofectamine 2000, and the stable intergrant was selected with G418 medium. The expression of enhanced green fluorescent protein (EGFP) protein was detected by fluorescence microscopy and EGFP/stathmin fusion protein by Western blot assay in transfected EC9706 cells. The growth curve of the two stably transfected cells was protracted with cell counting. FACS was used to detect the cell cycle. The clone formation rate in plate and in nude mice was tested to investigate the tumorigenic characteristics of the two stably transfected cells in vitro and vivo.

RESULTSA 450 bp coding sequence of stathmin cDNA was amplified by RT-PCR, which was cloned into pMD18-T vector. After identified with restriction enzyme the recombinant plasmid pMD18-T-stathmin containing reverse inserting sequence was constructed successfully. Then, the sub-clone pEGFP-stathmin was sequenced, confirming that the recombinant vector was right. The recombinant plasmid pEGFP-stathmin and pEGFP-C2 vector were transfected separately into EC9706 cells. After selecting with G418, the cells were transfected steadily. EGFP in EC9706 cells was observed after transfection by fluorescence microscopy. The expressed product was proved to be 46,000 EGFP/stathmin fusion protein by Western blot. Compared with those transfected with pEGFP-C2, the growth of cells transfected with pEGFP-stathmin became slow, the cells were swelled, the cell cycle was blocked at G2/M phase, the average clone formation rate decreased in vitro, and the tumorigenicity of inoculated cells in nude mice was decreased.

CONCLUSIONThe recombinant eukaryotic expression vector pEGFP-stathmin has been constructed successfully. It expresses steadily in esophageal cancer cells and inhibits the proliferation and tumorigenicity of transfected cells.

Animals ; Cell Cycle ; Cell Proliferation ; Escherichia coli ; genetics ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Genetic Vectors ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Plasmids ; Recombinant Fusion Proteins ; genetics ; metabolism ; Stathmin ; genetics ; metabolism ; Transfection