Objective To observe the effect of Danzhi Jiangtang Capsules(DJC) on serum interleukin-18 (IL-18),cystatin C (CysC) and relative biochemical indexes in patients with early diabetic kidney disease(DKD),and to explore its protective action on the kidney.Methods Sixty-two hospitalized patients with early diabetic nephropathy (period 11-Ⅲ) in Endocrinology Department of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine were divided into western medicine group and DJC group,31 cases in each group.The patients in western medicine group received conventional hypoglycemic drugs,and DJC group received DJC based on the treatment for the western medicine group.Before and after treatment,we observed the levels of serum IL-18,CysC,fasting plasma glucose (FPG),2-hour postprandial glucose (2hPG),glycosylated hemoglobin (HbA1c),serum creatinine (SCr),blood urea nitrogen (BUN),microalbumin (mAlb),urinary albumin /creatinine ratio (UACR),and estimated glomerular filtration rate (eGFR).Results The total effective rate was 90.0％ in DJC group and 76.7％ in the western medicine group,there being significant difference between the two groups(P ＜ 0.05).After treatment,the main traditional Chinese medical syndrome scores of DJC group were significantly lower than those before treatment,and the improvement was better than that of the western medicine group (P ＜ 0.05 or P ＜ 0.01).After treatment,the serum levels of IL-18,CysC,2hPG,SCr,BUN,mAlb,UACR in the two groups were significantly lower than those before treatment (P ＜ 0.05 or P ＜ 0.01),while eGFR level was higher than that before treatment (P ＜ 0.05 or P ＜ 0.01),and the improvement effect (except for BUN) in the DJC group was superior to that in the western medicine group(P ＜ 0.01).DJC had an effect on lowering the FPG and HbA1c (P ＜ 0.05 or P ＜ 0.01 compared with those before treatment),and the western medicine had no obvious effect on the two indexes (P ＞ 0.05).The results of Spearman correlation analysis showed that IL-18 was correlated with CysC,IL-18 and CysC were correlated with HbA1c,mALB and uACR.Conclusion DJC can protect renal function of patients with early diabetic nephropathy.

Objective To observe the sub-chronic effects of low doses of arsenic poisoning in rabbits exposed to different periods on some of the serum enzymes and biochemical indicators, and to provide the basis for screening of meaningful hematologic indicators for early diagnosis of arsenic poisoning. Methods Twelve adult rabbits,weighing 2.0 - 3.5 kg, were randomly divided into four groups, 3 in each group, and they were fed with drinking water containing sodium arsenite 0(control),0.01,0.05,0.25 mg/L, respectively. Serum alanine aminotransferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP), γ-glutamyl transacylase (y-GT), total protein(TP), albumin(ALB), globulin(GLP), and ALB/GLP of rabbit were measured by SYSMEX-180 automated biochemistry analyzer after 8 weeks and 12 weeks exposure. Results The results showed that ALT in 0.05 mg/Lgroup of 12 week[(60.00 ± 4.14)U/L]increased significantly compared with the control[(41.50 ± 2.12)U/L, P ＜0.05];AST in 0.25 mg/L group of 8 week and 12 week[(46.50 ± 3.21 ), (52.33 ± 3.81 )U/L]increased significantly compared with the control[(21.33 ± 3.53), (29.50 ± 3.23 )U/L, all P ＜ 0.05];ALP in 0.05 mg/L and 0.25 mg/L group of 12 week [(78.68 ± 4.85 ), ( 103.00 ± 7.83 ) U / L]increased significantly compared with the control [(45.50 ± 5.50)U/L, all P ＜ 0.05];γ-GT in 0.05 mg/L group of 12 week[(19.33 ± 7.50)U/L]increased significantly compared with the contro1[(8.50 ± 3.53)U/L, P＜ 0.05]. TP, ALB, GLP, ALB/GLP of different groups of 8 week and 12 week were not significantly different statistically(F= 0.77,0.02,0.16,3.14 and 0.51,0.29,0.41,0.52, all P ＞ 0.05). Conclusions Zero point zero five mg/L and higher doses of sub-chronic arsenic exposure has some major damage to the liver. Compared with other serum enzymes and the biochemical indexes, serum AST is a early sensitive indicator of liver injury of the arsenic poisoning.

Objective To observe the correlation between bone mineral density (BMD) and surgical outcomes of posterior lumbar interbody fusion (PLIF) for lumbar degenerative spondylolisthesis (DS).Methods From January 2006 to December 2010,69 patients with DS had undergone PLIF by the same surgical team.According the BMD,the cases were divided into two groups.Normal group (T ≥-1.0) had 33 cases [Male 16 cases,Female 17 cases; mean age,(56.5±9.0) yrs; L,,5 20 cases,L5S1 13 cases].The osteopenia group (T ＜-1.0) had 36 cases [Male 13 cases,Female 23 cases; mean age,(60.5±7.8) yrs; L4.5 21 cases,L5S1 15 cases].Blood loss,surgical duration,intra-and post-operative complications were collected.The clinical improvement was quantified by measurement of pain (visual analogue scale,VAS) and Roland-Morris (RM) Disability Questionnaire.Between two groups,the differences of age,body mass index,blood loss,VAS improvement,and RM improvement were compared.The correlation between BMD and sex,age,segment,screw loose,nonunion,and cage subsidence was analyzed.Results In two groups,the difference between pre-and post-operative RM and VAS was significant respectively.The blood loss was 415.5± 105.8 ml in normal group,significantly less than 528.3±128.7 ml in osteopenia group.There was no significant difference in the duration between normal group (169.7±44.3 min) and osteopenia group (176.4±42.6 min).The improvement of VAS and RM between two groups had no significant difference.There was a negative correlation between the BMD and blood loss (r=-0.407,P=0.001).The other surgical outcomes (surgical duration,VAS improvement,RM improvement,cage subsidence,nonunion,screw loose and etc.) had no correlation with BMD.Conclusion There is a negative correlation between the BMD and blood loss in DS patients managed by PLIF.BMD has no effect on other surgical outcomes.

Objective To study the effect of metformin on the growth of megakaryocytic leukemia cell line Dami and to explore the molecular mechanisms of the inhibitory effect of metformin on the proliferation of Dami. Methods The Dami cells were cultured and divided into control and 1,2,4,8,16 and 32 mmol·L-1 metformin groups.Then MTT test was performed to detect the inhitory rate of proliferation of Dami cells after treated with different concentrations of metformin. Flow cytometry was used to examine the distribution of cell cycle, and Western blotting was carried out to analyze the expressions of Cdc2 and CylinB1 and the phosphorylation of Cdc2. Results The MTT results showed that compared with control group,the inhibitory rates of proliferation of the Dami cells in 32 mmol·L-1 metformin groups at 0,24,48,72 and 96 h (35.1%±2.3%,49.7%±5.1%, 78.85±0.9%,79.1%± 3.0%%,and 85.2%± 3.2%)were significantly increased(P<0.01),Furthermore, after metformin treatment for 72 h,the inhibitory rates of proliferation of the Dami cells in 1,2,4,8,16 and 32 mmol·L-1 metformin groups were (33.8 ± 0.3)%,(51.9 ± 0.2)%,(59.4 ± 1.6)%,(65.5 ± 2.0)%, (75.5±0.9)%,and (79.1±3.0)%,respectively. Metformin inhibited the growth of Dami cells in a time-and dose-dependent manner. The flow cytometry results results revealed that compared with control group, the percentages of Dami cells in G2/M phase in 1,2 and 4 mmol·L-1 metformin groups were increased from (26.0± 0.5)% to (38.5 ± 1.5 )%, (48.4 ± 1.1 )%, and (58.2 ± 2.7 )%;there was significant difference in the percentages of Dami cells in G2/M phase between control group and 4 mmol·L-1 metformin group (P<0.01). Western blotting analysis showed that compared with control group, the expressions of Cdc2 and CyclinB were evidently reduced, the phosophorylation of Cdc2 at Tyr1 5 was up-regulated, and the phosphorylation at Thr1 6 1 was down-regulated.Conclusion Metformin can inhibit the growth of Dami cells and induce G2/M arrest,and its mechanism may be related to inhibiting the activation of Cdc2/CyclinB1 complex.

Objective To explore effect of transforaminal interbody fusion (TIF) in the treatment of thoracolumbar fracture.Methods From January 2009 to June 2010,11 patients with thoracolumbar fracture underwent TIF in our hospital.There were 7 males and 4 females,aged from 22 to 54 years (average,37.8±9.5 years).Fracture occurred at T11,12 in 2 cases,T12L1 in 5 cases,L1,2 in 2 cases,L2,3 in 1 case,and L3,4 in 1 case.According to AO classification,all 11 cases were rated as type B.The thoracolumbar injury classification and severity score (TLICS) ranged from 7 to 10 (average,9.2±0.87).According to Frankel grading system,there were 3 cases of grade B,4 cases of grade C,3 cases of grade D,and 1 case of grade E.The operative time,blood loss,and complications were evaluated.Functional recovery was evaluated by Frankel grading system.Radiographic results were obtained before surgery and during follow-up,and Cobb angle was measured.Results The operative time ranged from 130 to 170 minutes (average,147.3±11.9 minutes).The blood loss ranged from 180 to 650 ml (average,369.1±110.2 ml).All patients were followed up for 13 to 26 months (average,20.2±4.3 months).There was no complication during operation and follow-up.X-rays at final follow-up showed that there was no internal fixation failure such as screws break and cages shift.The improvement in Frankel grade was observed in 9 cases.The average Cobb angle improved from preoperative -10.3°±7.8° (range,-21.2° to 3.2°) to 1.2°±7.4° (range,-7.9° to 17.2°) at final follow-up.Conclusion The TIF is an effective method for treating thoracolumbar fracture,which shows good results in deformity correction,improvement of neurological function,bone fusion and stability of internal fixation.

Objective To investigate the efficacy and safety of anterior approach using an expandable artificial vertebral body for the correction of post-traumatic kyphosis (PTK) in the thoracolumbar spine.Methods From August 2009 to August 2011,13 patients with PTK in the thoracolumbar spine were treated through an anterior approach using an expandable artificial vertebral body.There were 4 males and 9 females,aged from 38 to 62 years (average,53.3±7.6 years).The injury levels consisted of T12 in 5 cases,L1 in 6 cases and L2 in 2 cases.All the operations were done by a single surgeon group.In the procedure,symptomatic vertebra and its two discs were excised,and the bony endplates were reserved.After putting an expandable artificial vertebral body into the space,the kyphosis was corrected by extending the artificial vertebral body.The operative duration,blood loss,Cobb angle,visual analogue scale (VAS) and Oswestry disability index (ODI) were recorded.Results All patients were successfully followed up for an average time of (18±5.5) months (range,12 to 28 months).The average Cobb angle was 33.9°±7.2°(range,22°to 53°)before operation and 7.3°±4.8°(range,2°to 16°)at final follow-up.The average VAS score was 6.4±0.9 (range,5 to 8)before operation and 1.5±0.8(range,0 to 3)at final follow-up.The average ODI was 50.5％±10.8％(range,38％ to 78％)before operation and 10.9％±4.9％(range,4％ to 22％) at final follow-up.All patients achieved bony fusion 12 months after operation.Conclusion Application of expandable artificial vertebral body through an anterior approach for PTK in the thoracolumbar spine has several advantages: large angle correction,less interruption of nerve,mini-invasion and less levels fixation.Satisfactory clinical outcome can be achieved.

1H NMR metabonomics approach was used to reveal the chemical difference of urine between patients with Xiao-Chaihu Tang syndrome (XCHTS) and healthy participants (HP). The partial least square method was used to establish a model to distinguish the patients with Xiao-Chaihu-Tang syndrome from the healthy controls. Thirty-four endogenous metabolites were identified in the 1H NMR spectrum, and orthogonal partial least squares discriminant analysis showed the urine of patients with Xiao-Chaihu Tang syndrome and healthy participants could be separated clearly. It is indicated that the metabolic profiling of patients with Xiao-Chaihu Tang syndrome was changed obviously. Fifteen metabolites were found by S-pot of OPLS-DA and VIP value. The contents of leucine, formic acid, glycine, hippuric acid and uracil increased in the urine of patients, while threonine, 2-hydroxyisobutyrate, acetamide, 2-oxoglutarate, citric acid, dimethylamine, malonic acid, betaine, trimethylamine oxide, phenylacetyl glycine, and uridine decreased. These metabolites involve the intestinal microbial balance, energy metabolism and amino acid metabolism pathways, which is related with the major symptom of Xiao-Chaihu Tang syndrome. The patients with Xiao-Chaihu Tang syndrome could be identified and predicted correctly using the established partial least squares model. This study could be served as the basis for the accurate diagnostic and reasonable administration of Xiao-Chaihu-Tang syndrome.
Humans ; Least-Squares Analysis ; Medicine, Chinese Traditional ; Metabolome ; Metabolomics ; Proton Magnetic Resonance Spectroscopy ; Syndrome ; Urinalysis

Objective To explore the sensitivity and the molecular mechanism of cisplatinresistance ovarian cancer cell line C13 to proteasome inhibitors and the combination with cisplatin. Methods After different treatments, methyl thiazolyl tetrazolium (MTT) assay was applied to examine the cell viability, annexin-V/propidium iodide(PI) apoptosis detection kit was used to determine the apoptosis rate of different groups, western blot assay was introduced to evaluate the expression levels of Fas-associated death domain-like interleukin-1 beta converting enzyme inhibitory protein (cFLIPs), and the activity of caspase-8 was examined. Results MTT assay shown that the cell viability ratios of combination group at serial time points from 12, 24, 36, 48, 60, 72 hours were ( 56.0 ± 8.4 ) %, (44.7 ± 7.3 ) %, ( 33.7 ±11.2) %, (27.6 ± 8.0) %, (27. 6 ± 7.6) % and (28.1 ± 2.4) %, which were much lower than those of cisplatin group (P <0.05). After treated for 24 hours, apoptosis rates of cisplatin group, bortezomib group and combination group were ( 16.7 ± 1.7) %, (23.4 ± 2.1 ) % and (26.9 ± 1.6) %, respectively. The rate of combination group was much higher than that of non-treated group and that of cisplatin group or bortezomib group ( P < 0.05 ). Western blot assay showed the changes of expression levels of cFLIPs, which were downregulated seriously after cisplatin, bortezomib or combination treatment [ (43.2 ± 2.3 )% vs( 75.7 ± 3.0)%vs (67.9 ± 2.1 ) %, P < 0.05 ]. The caspase-8 activity of combination group was (5.6 ± 1.6) folds than that of non-treated group, which was higher than those of other two groups [ ( 2.3 ± 1.0) and (4.2 ± 0.9 ) folds,P < 0.05 ]. Conclusions The tumor cell lethal effect of cisplatin could be increase significantly by the combination application of proteasome inhibitors, bortezomib. And the cFLIPs/caspase-8 signaling pathway may be play an important role in the molecular mechanism of the combination treatment.

Objective To detect the immune effect of FbaAmAb2 against the surface protein of group A Straptococcus (GAS),and explore the pathogenesis and therapy of GAS infections.Methods By subclonal and bacterial ELISA,the positive hybridoma cells were screened that can produce better titers of FbaAmAb2 against GAS-surface FbaA protein,and were injected into the peritoneal cavities of BALB/c mice to produce ascites.The collected ascites were performed to dilute,as follows,original ascite,1:2,1:4,1:8,and 1:16 to test tube agglutination.Based on the results,we selected appropriate dilution to passively immunize mice,and then challenged the mice with GAS,evaluating FbaAmAb2 neutralizing ability with GAS in mice by the survival rate of the immunized mice.Whether FbaAmAb2 could inhibit the binding of factor H to GAS was confirmed by the invasive inhibition assay.Results The IgG titer of bacteria solution ELISA is 1:160 and the titer of tube agglutination is 1∶8.The protect rates of FbaAmAb2 on preventing mice with GAS infections are as follows:66.67％ in original ascite and 1:2 diluted groups,and 50％ in 1:4 diluted group.Mice in each experimental group were evoked significantly protective immune responses compared with the PBS control by SPSS analysis.FbaAmAb2 can competitively inhibit factor H binding to the surface proteins FbaA of GAS,which decreased the entry of GAS into the cytoplasm of human epithelial cells through the binding of factor H.Conclusion FbaAmAb2 is promising to be used in emergent prevention or the clinical therapy for GAS infection and it is promising starting points for pharmacologic targeting and further development of new therapeutic agents for GAS.

Objective To explore the semitivity of ovarian cancer cell line SKOV3 to paclitaxel,oroteasome inhibitors,bortezomib,and their combination.Methods The methyl thiazolyl tetrazolitim (MTT)assay was applied to examine the cell viability after treatment.The annexin V-propidium iodide apoptosis detection kit was used to determine the apoptosis rate of different groups.Western blot assay was used to evaluate the expression levels of phosphorylated protein kinase B(AKT)and glycogen synthase kinase-3 beta(GSK-3β).Results In MTT assay,the cell viability ratios of the combination group at serial time points from 12,24,36,48 and 72 hours Were(65.2±5.8)%,(58.3±14.4)%,(35.3±5.0)%,(19.2±1.5)%,and(11.4 ±2.5)%,which were significantly lower than those of the paclitaxel group (P<0.05).After arug treatments,apoptosis rates of paclitaxel group,bortezomib group and the combination group were (14.7±0.5)%,(15.1±0.8)%and(20.5±0.7)%respectively.The rate of the combination group was significantly higher than that of non-treated group and paclitaxel group(P<0.05).Western blot assay showed the changes in expression levels of phosphorylated AKT and GSK-3β,which were decreased significantly after paclitaxd and bortezomib combination treatment [(3.2±0.8)%,(19.3±0.4)%;P<0.05].Conclusions The lethal effect of paclitaxel on tumor cells could be increased significantly by its combination with proteasome inhibitors,bertezomib.The AKT/GSK-3β signaling pathway plays an important role in the molecular mechanism of the combination treatment.