ObjectiveTo observe the clinical efficacy of qi-blood-supplementing needling in treating the incipient leukopenia after chemotherapy for breast cancer.MethodSeventy-eight patients with incipient leukopenia after chemotherapy for breast cancer were recruited and randomized into a control group (38 cases) and a treatment group (40 cases). The control group was intervened by medications for increasing white blood cell (WBC) count, while the treatment group was by qi-blood-supplementing needling plus the medication. The therapeutic efficacies were evaluated at the end of the intervention.ResultThe WBC counts increased significantly in both groups after treatment (P<0.01), and the count in the treatment group was significantly higher than that in the control group (P<0.05). The total effective rate was 95.0% in the treatment group versus 89.5% in the control group, and the therapeutic efficacy of the treatment group was superior to that of the control group (P<0.05). The treatment group was also better than the control group in improving symptoms including poor appetite, fatigue, pale complexion, and lassitude (P<0.05,P<0.01).ConclusionFor patients with incipient leukopenia after chemotherapy for breast cancer, qi-blood-supplementing needling plus medication for increasing WBC count can up-regulate the WBC count and improve the symptoms due to qi-blood deficiency.

BACKGROUND: Some studies suggest that pre-injection of tumor necrosis factor-α (TNF-α)can protect focal cerebral ischemia in mice. Cerebral ischemia tolerance is related to the increase of TNF-α level; on the other hand, TNF-α is an injurious cytokine associated with stroke. Circulating antibody against anti-TNF-α can protect reperfused injury.OBJECTIVE: To study the effects of TNF-α pretreatment and post-treatment on cerebral ischemia-reperfusion injury and explore possible mechanism.DESIGN: Randomized controlled study.SETTING: Neurological Department, the Second Hospital Affiliated to Harbin Medical University.MATERIALS: The experiment was conducted at the Animal Experiment Center of Harbin Medical University from January to April 2002. Totally 120 healthy adult male Wistar rats were randomly divided into the following 8 groups: TNF-α 0.05 μg, 0.5 μg and 1.0 μg pretreatment groups and PBS group, TNF-α 0.05 μg, 0.5 μg and 1.0 μg post-treatment groups and PBS group with 15 in each group.METHODS: The focal brain ischemia model of middle cerebral artery occlusion (MCAO) was made using inserting thread method. TNF-α of different doses (0.05 μg, 0.5 μg or 1.0 μg) or PBS was injected intracisternally and 22-hour reperfusion, 8 rats from each group were killed. Then the perhour reperfusion, 7 rats from each group were killed. Then pathological changes were observed, glial fibrillary acidic protein (GFAP) and intercellular adhesion molecule-1 (ICAM-1) expression were inspected by immunohistochemical method. Histopathological and immunohistochemical evaluation was made with the computer-assisted image analyzing system,and the number of GFAP positive cells and ICAM-1 positive vessels in each hemisphere was counted.riliary acidic protein and ICAM-1.infarct volume: TNF-α 0.5 μg and TNF-α 1.0 μg pretreatment groups showed reduced volume of lesion; infarct volume reduced by 70.9% in TNF-α 0.5 μg pretreatment rats and 66.5% in TNF-α 1.0 μg pretreatment rats. TNF-α 0.5 μg and TNF-α 1.0 μg post-treatment groups showed increased volume of lesion; infarct volume increased by 22.3% in TNF-α 0.5 μg post-treatment rats and 46.7% in TNF-α 1.0 μg post-treatment rats.TNF-α 0.05 μg and 1.0 μg pretreatment groups did not differ significantly (P ＞ 0.05), but there was an obvious difference between TNF-α 0.5 μg and pared with PBS pretreatment group, TNF-α 0.5 μg and 1.0 μg pretreatment groups showed lessened tissue damage and edema. Compared with PBS post-treatment group, TNF-α 0.5 μg and TNF-α 1.0 μg post-treatment fibriliary acidic protein and ICAM-1: TNF-α 0.5 μg and TNF-α 1.0 μg pretreatment groups showed reduced volume of glial fibriliary acidic protein and ICAM-1 (P ＜ 0.05); but TNF-α 0.5 μg and TNF-α 1.0 μg posttreatment groups showed increased volume of glial fibriliary acidic protein and ICAM-1 (P ＜ 0.05). TNF-α 0.05 μg and 1.0 μg pretreatment groups did not differ significantly (P ＞ 0.05); but there was an obvious difference between TNF-α 0.5 μg and 1.0 μg post-treatment groups (P ＜ 0.05).cerebral ischemia reperfusion injury. This effect is not related to the repair given after cerebral ischemia reperfusion, ischemia exacerbates, which is α are determined by whether TNF-αis given before or after cerebral ischemia in a dose-dependent manner.

A novel high performance liquid chromatographic method was developed for the determination of 4-O-methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation.Plasma samples were treated by one-fold volume of methanol and acetonitrile to remove the interference proteins.A reverse phase column of SHIMPACK VP-ODS (150 mm× 4.6 mm,5.0 μm) was used to separate 4-O-methylhonokiol in the plasma samples.The detection limit of 4-O-methylhonokiol was 0.2 μg/L and the linear range was 0.012- 1.536 mg/L.The good extraction recoveries were obtained for the spiked samples (84.7％,89.3％ and 87.7％ for low,middle and high concentrations of added standards,respectively).The relative standard deviation of intra-day and inter-day precisions was in the range from 0.6％ to 13.5％.The pharmacokinetic study of 4-O-methylhonokiol was made and the results from the plasmaconcentration curve of 4-O-methylhonokiol showed a two-apartment open model.This work developed a sensitive,stable and rapid HPLC method for the determination of 4-O-methylhonokiol and the developed method has been successfully applied to a pharmacokinetic study of 4-O-methylhonokiol.

Objective To detect the influence of marOR mutations on antibiotic resistance in Shigella spp. Methods The marOR gene with four-base deletion was amplified by overlap PCR, then inserted in a T-vector and transformed into DH5α. The clone of marOR gene with four-base deletion and three point mutations was prepared from the strain having these mutations. Electrophoresis and sequencing were preformed to certify the correction of the cloned genes. Drug susceptibility tests were preformed for the strains harbouring the different clones [DH5α, DH5α (T), DH5α (marOR), DH5α (marOR-CATT), DH5α(marOR-CATT + 3m)]. Results Compared with the control strain (DH5α-T), the antibiotic resistances of marOR with four-base deletion [DH5α (marOR-CATT)] were higher to streptomycin, tobramycin, cefazolin and cefalexin, and the antibiotic resistances of marOR with four-base deletion and three point mutations [DH5α (marOR-CATT + 3m)] were higher to streptomycin and to tetracycline. The antibiotic resistances of DH5α (marOR-CATT) and DH5α (marOR-CATT +3m) to streptomycin, tetracycline, chloramphenicol, cefazolin, levofloxacin, ciprofloxacin and norfloxacin were higher than DH5α (marOR). The diameters of the antibiotics except the trimethoprim between DH5α (marOR-CATT) and DH5α (marORCATT +3m) had not significant disparity. Conclusion The four-base deletion in 1376-1379 sites of the marOR gene increased the resistance of Shigella spp to some antibiotics. The point mutations in 1411, 1417,1435 sites of the marOR gene have little influence on the antibiotic resistance of Shigella spp.

Objective To understand epidemic characteristics of human influenza A and the genetic and antigenic variations of H1N1 influenza A isolates in Shanghai area in 2009. Methods Throat swabs were collected from patients with influenza-like illness in the sentinel surveillance clinic in Shanghai area in 2009, then inoculated in Madin-Darby canine kidney (MDCK) cell lines. The types of influenza were identified by direct immunofluorescence assay (DIF) and the subtypes were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Segments of hemagglutinin (HA) and neuraminidase (NA) genes of some 2009 H1N1 influenza A isolates were amplified and sequenced. HA and NA gene mutations of 2009 H1N1 influenza A isolates were analyzed. Results Seasonal H1N1 and H3N2 influenza A viruses co-circulated during the spring of 2009 in Shanghai area. Seasonal H3N2 began to co-circulate with 2009 H1N1 in August (the 32nd week) and finally2009 H1N1 became dominate since the 40th week. The phylogenetic tree of 2009 H1N1 HA segment revealed that the isolates from different regions and months were interspersed with each other, but all were clustered into one branch which closed to strains in Spain, Russia, Denmark and other European countries. Mutations were found in some HA amino acid sites, but none of them was in the antigenic determinant region. No change was observed in the 274 NA amino acid residues which were related to the drug resistance to oseltamivir. PB2 protein analysis showed that the 627 and 701 amino acid residues were glutamic acid and aspartic acid respectively, which were the same encoded amino acid with avian flu PB2 protein. Conclusions Seasonal H1N1 and H3N2 co-circulated in the spring of 2009, then both 2009 H1N1 and seasonal H3N2 were prevalent in Summer and Autumn, and 2009 H1N1 finally became dominate in Autumn. Compared to early 2009 H1N1 strains, variations are detected in H1N1 influenza A viruses, but none of them has epidemiological influence, and viruses still show high affinity with human and low-pathogenic characteristics.

Objective To observe therapeutic effects of HuGanJieXian decoction on rats hepatic fibrosis induced by tetrachloride. Methods Rat models of hepatic fibrosis were constructed by intraperitoneal injection of tetrachloride.HuGanJieXian decoction composed of low, middle, and high dose curcumin were given to these rats respectively at the same time. Sho-saiko-to compound treatment group and Fufangbiejiarangan Tablets treatment group were made as positive control groups. After twelve weeks, all rats were executed. Serum samples were kept for measuring serum levels of PC-Ⅲ, LN, and HA. Left livers were extirpated for pathologic examination including H.E and Masson stainings. Grade of hepatic fibrosis were evaluated according to SSS system. The levels of superoxide dismutase (SOD) and malondialdehyde (MDA) of supematant centrifugated from hepatic tissue homogenate were detected. Results Serum levels of PC-Ⅲ, LN, and HA were depressed obviously in decoction groups compared with those of fibrotic group (P<0.05) , especially in the low-dose curcumin group.HuGanJieXian Decoction could increase the level of SOD and decrease the level of MDA (P<0.05) , especially in the low-dose curcumin group. Staining of H. E and Masson showed that degrees of hepatic fibrosis in decoction groups were improved obviously compared with that of the fibrotic group. Conclusion HuGanJieXian Decoction can improve rat hepatic fibrosis, the mechanism of this effect may be associated with protecting hepatic cell membrane and anti- peroxidative damage.

Objective To explore the value of virtual touch tissue quantification(VTQ) on the early diagnosis of gouty nephropathy.Methods 180 healthy controls and 109 gout patients including 57 cases with normal renal function,and 52 cases with gouty nephropathy were measured by VTQ in this study.The shear wave velocity (SWV) of renal cortex,renal medulla and renal sinus were measured using virtual touch tissue quantification(VTQ).The recorded SWV value was compared.Results In all groups,the SWV of renal cortex was the highest with significant difference (P ＜0.01).The SWV of renal cortex in gouty nephropathy group was higher than those in control group (t =0.342,P =0.026).The SWV of renal medulla and sinus in gout with normal renal function group and gouty nephropathy group were higher than those in control group,respectively (gout with normal renal function group:t =-0.311,-0.628; P =0.012,0.000;gouty nephropathy group:t =-0.369,-0.701 ; P =0.000,0.000).The SWV of renal cortex,medulla and sinus had no significant difference between gout patients with normal renal function and gouty nephropathy patients(t =0.117,-0.059,-0.073; P =0.232,0.575,0.523).Conclusions The VTQ technology could quantitatively evaluate tissue elasticity of the gout patients and provide evidence for the early diagnosis of gouty nephropathy.

Objective To explore the relationship between tubal intraepithelial carcinoma (TIC) of the fimbria and pelvic high-grade serous carcinoma.Methods All 34 cases of pelvic high-grade serous carcinoma with clear fimbria were evaluated from January 2009 to June 2010,including ovarian carcinoma (n=26),tubal carcinoma (n=7) and peritoneal carcinoma (n=1).Among of these ovarian carcinomas,12 cases were surface deposits and the other 14 cases within ovarian parenchyma.All 42 cases of non highgrade serous carcinoma in this period including 13 endometrioid ovary carcinomas,11 clear cell ovary carcinomas,11 mucinous ovary carcinomas,6 low-grade serous ovary carcinomas,1 low-grade serous tubal carcinoma,were also collected as a reference.The presence of tubal intraepithelial carcinomas was assessed.Based on the presence of TIC,high-grade serous ovary carcinomas were divided into TIC positive (+) and TIC negative (-) groups,and the clinical and pathological features of them were also evaluated.Results Fifteen cases (44％) were identified TIC in 34 high-grade pelvic serous carcinomas,and all of them were in the fimbria only,while none of TIC was found in control cases.There were significant difference between the two groups (x2=23.086,P=0.000).Eleven cases(42％) were identified TIC in all 26 high-grade ovarian serous carcinomas,in which 8 cases with unilateral ovary carcinomas were associated with ipsilateral TIC,2 cases with bilateral ovary carcinomas associated with unilateral TIC and one case with bilateral ovary carcinoma was associated with bilateral TIC.Four TIC (4/7) were identified in 7 cases with high-grade tubal serous carcinomas,and there was no presence of TIC in the 1 high-grade serous peritoneal carcinoma.Of all 26 high-grade ovarian serous carcinomas,6/11 cases were surface deposits,and 5/11 were parenchyma tumors in TIC (+) group while 6/15 cases were surface deposits and 9/15 were parenchyma tumors in TIC (-) group,in which there were correlated in distribution of TIC between the two groups( P＞0.05 ).The average diameter of ovarian cancer were 6.9 and 6.5 cm between the two groups with no significant differences ( t=0.409,P=0.690).Conclusion TIC is specific to high-grade serous carcinomas and maybe have something to do with the pathogenesis of pelvic serous carcinomas.

Objective To explore the application value of virtual touch tissue quantification(VTQ) technique in differentiating small solid thyroid nodules.Methods The patients with suspiciously malignant thyroid nodules(solitary,size range 5-15 mm),homogeneous and normal color Doppler flow imaging in the rest of thyroid parenchyma,were enrolled in this study.Five repeated measurements were performed on both nodules and extra-nodular tissues using VTQ.A receiver-operating characteristic (ROC) curve was drew according to shear wave velocity(Vs).Results 62 patients with a total of 67 lesions were included in this study,turned out to be 39 malignant and 28 benign.The Vs of malignant group nodules,malignant group extra-nodular tissues,benign group nodules and benign group extra-nodular tissues were (3.91 ±1.85) m/s,(2.03 ± 0.42)m/s,(2.29 ± 0.56)m/s,(1.84 ± 0.45)m/s,respectively.The Vs difference between malignant and benign nodules was statistically significant (P =0.000),while the Vs difference between malignant and benign extra-nodular tissues had no statistical significance (P =0.07).The best cutoff Vs value for distinguishing malignant from benign nodules was 2.78 m/s,with the sensitivity,specificity and accuracy of 76.9％,78.6％ and 80.6％ respectively.The area under ROC curve was 0.84.Conclusions VTQ could evaluate the elasticity of thyroid nodules,and was helpful in the diagnostic work-up of thyroid nodules.

AIM: To investigate the feasibility and its mechanisms of improving therapeutic effect by antisense gene therapy combined with chemotherapy in osteosarcoma. METHODS: The human osteosarcoma implanted tumor model in the nude mice was established. By intratumoral injection and abdominal cavity administration, the tumor bearing mice were treated with survivin ASODN in combination with diamminedichloroplatinum (DDP) for a week. Comparison with each single - agent therapy and control group was performed in aspects such as tumor growth condition, pathological changes of tumor tissues; survivin protein expression in tumor tissues by immunohistochemistry, survivin mRNA expression levels by RT -PCR method and tumor apoptosis by Tdt -mediated dUTP nick end labeling (TUNEL). RESULTS: All nude mice survived the therapy. As compared with the control group, the antisense gene therapy group presented synchronous decrease in survivin mRNA and protein expression; all therapy group displayed tumor growth inhibition and cell apoptosis with different extent; while in contrast to single - agent therapy group, the combined therapy group showed stronger inhibition of tumor growth and abundant tumor cell apoptosis with the highest apoptotic rate. CONCLUSION: Synergistic effect was achieved by combination of DDP with ASODN that may overcome drug resistant of DDP and the combined strategy may shed new light on the cancer therapy.