Objective To investigate the clinical therapeutic effects of traditional Chinese medicine on different stages of nontraumatic necrosis of femoral head.Methods The clinical data were retrospectively analyzed of 359 patients(239 males and 120 females,aged 16-69 years,averaged 40.6) suffering from nontraumatic necrosis of femoral head,admitted to our hospital from Aug.2001 to Aug.2008,and treated with traditional Chinese medicine.The patients were divided into five stages(0,1,2,3,and 4) according to Huang Changlin's and Ficat's clinical diagnostic criteria.The age,gender and disease course of patients in different stages were of comparability.All the patients were treated with traditional Chinese medicine for 3-6 months and followed up for 1-2 years,and the final therapeutic effects were evaluated by the comprehensive evaluation criteria.Results The clinical cure rates in patients of stages 0,1,2,3 and 4 were 44.9%,36.1%,30.5%,0% and 0%,and the effective rates were 97.8%,80.6%,75.6%,63.6% and 50.0%,respectively.The total cure rate was 25.3%,and the total effective rate was 76.3%.The therapeutic effect in patients of stage 0 was better than those of stages 1 and 2(P0.05).Conclusions The therapeutic effect of traditional Chinese medicine is remarkable on the patients with nontraumatic necrosis of femoral head in stages 0,1 and 2,but not in stages 3 or 4.Traditional Chinese medicine may increase the cure rate of early stage patients,and improve the symptoms of advanced stage patients.

Objective:To eliminate the bacteriostatic action ofA,wei Capsule in microbial limit examination so as to establishing the method ofmicrobial limit for A,wei Capsule.Methods:The plating and culture medium diluted methods were used to determine the antimicrobial effect ofA,wei Capsule by recovery rates with 5 control trains.Results:According to the results, the recovery rates were higher than 70% by culture medium diluted method, which indicated no antimicrobial effect with them.Conclusion:According to the results, the culture medium diluted method can eliminate the bacteriostatic action ofA,wei Capsule preferably with accurate result and reaching the objective.

Objective:To elucidate the possible role of galanin in the development of experimental depression in rats. Methods:Openfield was performed to test the behavior of rats. The changes of the galanin level in different brain areas were determined by RIA. The effect of fluoxetine hydrochloride on galanin level were observed by intraperitoneal injection. Results:Compared to control group, the crossing times and rearing times decreased significantly in depressed rats, galanin level decreased remarkably in plasma, hypothalamus, hippocampus, forebrain, parietal lobe and temporal cortex of depressed rats. Intraperitoneal injection of fluoxetine hydrochloride obviously improved the depressed behavior in rats, increased the galanin level in the hippocampus and forebrain of depressed rats. Conclusion:Hippocampus and forebrain may be involved in the development of experimental depression and in the antidepressive effects of fluoxetine hydrochloride.

Objective To observe the month age distribution of peroxisome proliferators activated receptor gamma (PPARγ) expression in rat kedney. Methods Wistar rats aged 3 months,12 months and 24 months were made as models who represented young, middle-aged and old group respectively. Western blotting, immunohistochemical (IHC) and in-situ hybridization (ISH) were used to detect the expression and location of protein and mRNA of PPARγ in rat kidney. Results Western blotting results showed that the expression of PPARγ protein was higher in 3 months group than in 24 months group (0.94±0.05 vs. 0.78±0.02, P＜0.01) and 12 months group (0.87±0.04, P＞0.05), and it reduced in 24 months group than in 12 months group (P＞0.05). By IHC,the PPARγ protein was localized predominantly in the nuclear of tubular epithelia and collecting duct cells in each group. In old age group, PPARγ protein was also detected little in the mesangial and Bowman's capsule epithelial cells. Meanwhile, the distribution of PPARγ mRNA with ISH was consistent with above findings. Additional, semi-quantitative analysis of ISH results verified that the level of PPARγ mRNA decreased with ageing. Conclusions As a nuclear transcription factor,PPARγ participates in the regulation of rat kidney aging.

Objective To investigate the effect of rosiglitazone (RGTZ) on anti-oxidation in aging rat kidney. Methods Twenty-four-month-old male Sprague-Dawley rats were randomly divided into three groups (n=10): control group (CON), rosiglitazone group (RGTZ) and caloric restriction group (CR). The CON rats were allowed ad libitum access to feed and tap water.The RGTZ rats received intragastric administration of RGTZ (4 mg·kg-1·d-1),and the CR rats were provided with a vitamin and mineral fortified version of the same diet at a level of 40% less food (by weight) than the CON rats. After 12 weeks all the animals were sacrificed by decapitation, and both the body weight and the percentage of kidney and heart in each group were measured.Western blot was performed to analyze the expression of PPARγ protein. The content of MDA and the activity of SOD and GSH-PX in kidney tissue were detected. Besides, frozen sections of kidney tissue were stained for senescence-associated-13 galactosidase (SA-β-Gal). Results The body weight of CR rats decreased obviously, in contrast, which did not change in CON and RGTZ group. Percentage of kidney and heart to body weight was normal in CR or RGTZ group after intervention. Western blot result showed that PPARγ protein expression in rat kidney was significantly higher in RGTZ and CR group as compared to CON group (P<0.05). Compared with RGTZ and CR rats, obviously lower activities of SOD and GSH-Px were noted in CON rats, however, the content of MDA was higher in CON rats. Additionally, the positive staining area of [3-Gal in CR and RGTZ group was significantly smaller than that in CON rats (P<0.05, P<0.01 ). Conclusion RGTZ can defer the kidney aging in senescence SD rat, and the mechanism may be related to amelioration of oxidative damage and enhancement of antioxidation.

Objective:To examine the roles of follicular helper T(Tfh)cells,serum IL-21 and B cells in the pathogenesis of alcohol abuse non-traumatic osteonecrosis of the femoral head ( NONFH ) .Flow cytometry was used to measure the frequencies of peripheral blood inducible Tfh cells and B cells in alcohol abuse NONFH patients and healthy controls.The disease progression and the extent of femoral head collapse,the serum IL-21 were quantified.Methods: A significantly higher percentages of CD19+B cells(t=3.765,P=0.0005),CD86+CD19+B cells(t=5.506,P<0.0001),and CD95+CD19+B cells(t=4.152,P=0.0002) in patients than those in controls was found.The percentages of CD86+CD19+B cells were positively associated with the index of femoral head collapse in alcohol abuse NONFH(P<0.0001,r=0.536).Results: The frequencies of Tfh cells (t=7.611,P<0.0001),and IL-21+Tfh cells (t=5.281,P<0.0001) were higher than those in controls;The frequencies of Tfh cells were positively associated with the percentages of CD19+B cells(P=0.0002,r=0.455),IL-21+Tfh cells were positively associated with the percentages of CD86+CD19+B cells(P=0.0002,r=0.447).Conclusion: Tfh cells and B cells may participate in the pathogenesis of alcohol abuse NONFH,and increased CD86+CD19+B cells may be associated with the extent of femoral head collapse,the interaction of Tfh cells and B cells may have an im-portant role in pathogenesis of alcohol abuse NONFH.

Objective To develop a machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding and to evaluate its effectiveness of field application,so as to provide a novel Oncomelania hupensis snail control technique in the large?scale marshlands. Methods The machine simultaneously integrating mechanized environmental clean?ing and automatic mollusciciding,which was suitable for use in complex marshland areas,was developed according to the mech?anization and automation principles,and was used for O. hupensis snail control in the marshland. The effect of the machine on environmental cleaning and plough was evaluated,and the distribution of living snails was observed at various soil layers follow? ing plough. The snail control effects of plough alone and plough followed by mollusciciding were compared. Results The ma?chine could simultaneously complete the procedures of getting vegetation down and cut vegetation into pieces,plough and snail control by spraying niclosamide. After plough,the constituent ratios of living snails were 36.31% ,25.60% ,22.62% and 15.48% in the soil layers at depths of 0-5,6-10,11-15 cm and 16-20 cm respectively,and 61.91% living snails were found in the 0-10 cm soil layers. Seven and fifteen days after the experiment,the mortality rates of snails were 9.38% and 8.29% in the plough alone group,and 63.04% and 80.70% in the plough + mollusciciding group respectively(c27 d = 42.74,c215 d =155.56,both P values < 0.01). Thirty days after the experiment,the densities of snails were 3.02 snails/0.1 m2 and 0.53 snails/0.1 m2 in the soil surface of the plough alone group and the plough + mollusciciding group,which decreased by 64.92% and 93.60% ,respectively,and the decrease rate of snail density was approximately 30% higher in the plough + mollusciciding group than that in the plough alone group. Conclusions The machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding achieves the integration of mechanical environmental cleaning and automatic niclosamide spraying in the complex marshland areas,which provides a novel technique of field snail control in the large?scale setting in Chi?na.

Objective To screen Oxalobacter formigenes (OxF) from fresh feces of healthy adults,and study its effect on the the prevention of calcium oxalate kidney stones.Methods OxF was screened and cultured from fresh feces of healthy adults.The rat model of calcium oxalate stone was established by esophageal gavage of 0.8％ of ethylene glycol.Rats were divided into a control group and four groups of rats with ethylene glycol-induced calcium oxalate kidney stones according to random number table.Three groups were treated with 106 CFU,107 CFU,108 CFU viable OxF every day,respectively,for 4 weeks.The blood and 24-hour urine samples were collected to detect the serum creatinine,urea nitrogen,serum and urine calcium,phosphorus,magnesium and urine oxalate every week.At the end of the 4th week,the rats were sacrificed and the kidney tissues were stained with HE and Yasue.The deposition and content of calcium oxalate crystals were observed under a light microscope.Results The bacteria strain isolated from fresh feces of healthy adults was 100％ as same as the known ATCC35274 bacteria strain,which means the strain screened is OxF.Among the 5 groups,there were no significant differences in body weight,Scr,BUN,serum calcium,blood magnesium,blood phosphorus,urinary magnesium and urinary phosphorus.The 24-hour urinary calcium excretion in the model group was significantly lower than that of the control group (P ＜ 0.05).After intervention with OxF solution,the 24-hour urinary calcium excretion in the 108 CFU OxF group was significantly higher than that in the model group (P ＜ 0.05),while there was no significant difference between the other intervention groups and the model.The oxalic acid excretion of 106 CFU OxF group and 107 CFU OxF group was lower than that of the model,but the difference did not reach statistical significance (P＞ 0.05).The 24 h oxalic acid excretion in the 108 CFU OxF group was significantly lower than that of the model at the end of first week (P ＜ 0.05),and continued to decrease for the next 3 weeks.After 4 weeks of intervention,no crystal formation was observed in the control group under the deflection microscope,but a large amount of calcium oxalate crystals were formed in the renal cortex and renal medulla.The crystals were piled up and connected to each other.Yasue staining coincided with the calcium oxalate crystal in the same part of the kidneys.Compared with the model,there was no significant change in the score of calcium oxalate crystal in the kidneys of 106 CFU OxF group and 107 CFU OxF group,while the score of calcium oxalate crystal in the kidneys of 108 CFU OxF group was significantly lower (P ＜ 0.05).Conclusions OxF are successively screened from healthy adults.Daily administration of 108 CFU OxF can safely and effectively reduce the urinary oxalic acid excretion,prevent the formation of calcium oxalate crystals and inhibit the formation of stones in kidneys of rats.

Objective To explore the effect of miR-1-3p on the malignant biological behavior of human esophageal squamous cell carcinoma cells and the potential mechanisms.Methods The Gene Expression Omnibus(GEO)database was analyzed to screen differentially expressed miRNAs in esophageal squamous cell carcinoma(ESCC).qRT-PCR was used to detect the expression of miR-1-3p in human ESCC cell lines(KYSE30,KYSE150,KYSE410,KYSE510,and Eca109)and normal esophageal epithelial cell line HET-1A.CCK-8,wound healing,Transwell assays,and flow cytometry were applied to detect the effect of miR-1-3p on the proliferation,migration,invasion,and apoptosis of ESCC cells.Bioinformatics tool was used to predict the target genes of miR-1-3p.A Kaplan-Meier survival curve was drawn to analyze the correlation between STC2 expression and overall survival of patients in the ESCC cohort of the TCGA database.Fluorescence in situ hybridization was performed to verify the subcellular location of miR-1-3p in ESCC cells,and dual-luciferase reporter gene assay was performed to validate the regulation of miR-1-3p on stanniocalcin 2(STC2).RNA immunoprecipitation assays were used to detect the binding of miR-1-3p and STC2.Western blot assay was performed to determine the effect of miR-1-3p on the expression of STC2 and endoplasmic reticulum stress pathway-related proteins,including p-PERK,p-eIF2α,and ATF4.CCK-8,wound healing,Transwell assays,and flow cytometry were applied to detect the effect of STC2 overexpression and knockdown on the proliferation,migration,invasion,and apoptosis of ESCC cells.Results The expression of miR-1-3p was lower in ESCC cell lines than in HET-1A cells(all P<0.05).The transfection of miR-1-3p mimic decreased the proliferation,invasion,and migration of ESCC cells(all P<0.05)and promoted the apoptosis of ESCC cells(all P<0.001).Bioinformatics tool showed that STC2 was a target gene of miR-1-3p.The expression of STC2 in ESCC tissues was higher than that in normal esophageal epithelial tissues in the ESCC cohort of TCGA database and was negatively correlated with prognosis(all P<0.05).miR-1-3p was located in the cytoplasm and can directly bind to STC2 mRNA.The transfection of miR-1-3p mimic downregulated the expression of STC2,p-PERK,p-eIF2α,and ATF4(all P<0.05).The overexpression of STC2 promoted the proliferation,invasion,and migration(all P<0.05)and inhibited the apoptosis of ESCC cells(all P<0.05).Knockdown of STC2 inhibited the proliferation,invasion,and migration(all P<0.05)and promoted the apoptosis of ESCC cells(all P<0.05).Conclusion miR-1-3p inhibits the malignant biological behavior and promotes the apoptosis of esophageal squamous cell carcinoma cells by regulating STC2 possibly by suppressing the endoplasmic reticulum stress.

We constructed the CS1-targeted second- and third-generation CAR-T cells with genetic engineered 4-1BB or/and ICOS as a costimulatory signaling molecule by use of lentiviral platform. The CS1-targeted second-generation CAR-T cells with ICOS or 4-1BB had similar anti-neoplastic activity. When effector/target ratio was 1:1, the CAR-T cells with ICOS showed better killing effect on IM9-lucgfp cells than those with 4-1BB. However, The CS1-targeted third-generation CAR-T cells exihibited lower cytolytic capacity against IM9-lucgfp cells than the CS1-targeted second-generation CAR-T cells when the ratio of effector/target was 1:1, 2:1 or 5:1. When the ratio of effector/target was 10:1, the killing efficacy of both the second- and third-generation CAR-T cells against IM9-lucgfp cells was more than 85%, significantly higher than that of the control T cells. Taken together, both the CS1-targeted second- and third-generation CAR-T cells with ICOS or/and 4-1BB could efficiently kill CS1-positive multiple myeloma cells, but the CS1-targeted second-generation CAR-T cells had more potent killing effect on CS1-positive multiple myeloma cells than the CS1-targeted third-generation CAR-T cells.
4-1BB Ligand/metabolism* ; Cell Line, Tumor ; Genetic Engineering ; Humans ; Inducible T-Cell Co-Stimulator Protein/metabolism* ; Multiple Myeloma/therapy* ; Signal Transduction ; T-Lymphocytes/chemistry* ; Xenograft Model Antitumor Assays