Objective To study the effect of 2?-(3-hydroxypropoxy)-Calcitriol (ED-71) on the cell growth and differentiation of human osteoblast-like cell line OS-732. Methods Osteoblast-like cells were cultured with various concentration of ED-71.The effects of ED-71 on proliferation and differentiation were detected by (MTT), alkaline phosphatase activity and level of osteocalcin (bone gla protein,BGP). Results The proliferation rate of osteoblast-like cells cultured for 4 days was significantly inhibited by 10~ -7mol/L concentration of ED-71. ED-71 in 10~ -8, 10~ -9mol/L concentration can significantly increase BGP level, but ED-71 in 10~ -7mol/L concentration may decrease BGP level.Alkaline phosphatase activity compared with control was not changed significantly. Conclusions ED-71 can inhibit cell growth in high concentration and improve cell differentiation in lower concentration.

Objective:To observe the effect of acupuncture plus medium-frequency electric stimulation on bladder function after radical hysterectomy for cervical cancer. Methods:A total of 170 patients confirmed by biopsy and surgical pathology and treated with radical hysterectomy for cervical cancer were randomly divided into an acupuncture group, an observation group and a control group by random digital table. Fifty-six cases in the acupuncture group were treated with acupuncture after surgery, one session each day, till the extubation day; 58 cases in the observation group were treated with acupuncture plus medium-frequency electric stimulation after surgery, one session each day, till the extubation day; and 56 cases in the control group didn't receive any intervention, and the catheter was retained till the extubation day. On the postoperative day 14, all groups were dealt with catheterization to determine the residual urine volume, in order to observe the occurrence rate of urinary retention and the recovery of bladder function and to compare the differences of the therapeutic effects among the groups. Results:The occurrence rates of urinary retention were respectively 16.1%, 20.7% and 46.4% in the acupuncture group, observation group and control group, with significant differences (P<0.05). When comparing the acupuncture group and the observation group with the control group, there were significant differences (bothP<0.05). In comparison between the acupuncture group and the observation group, there was no significant difference (P>0.05). The recovery rates of bladder function in the three group were respectively 42.9%, 60.3% and 41.1% without significant differences (P>0.05). Moreover, there were no significant between-group differences (P>0.05), but acupuncture plus medium-frequency electric stimulation showed a better tendency. Conclusion: The single application of acupuncture therapy is effective and able to prevent urinary retention after radical hysterectomy for cervical cancer. Acupuncture plus medium-frequency electric stimulation can effectively enhance the recovery of bladder function and reduce the residual urine volume in the bladder to the maximum.

Objective To analyze locations of acantholysis in pemphigus vulgaris (PV) and pemphigus foliaceus (PF),so as to explain why acantholysis in pemphigus occurs in different locations of the epidermis.Methods Clinical data,histopathological and immunopathological findings,and pemphigus antibody level values were collected from 43 patients with PV and 28 with PF,and retrospectively analyzed.Results Of the 43 patients with PV,35 showed acantholysis in the upper basal layer,8 in the middle-to-upper epidermis.Of the 28 patients with PF,25 showed acantholysis in the granular layer and the upper prickle cell layer,3 in the middle-to-lower epidermis.Patients with PF showed significantly higher levels of anti-Desmoglein 1 (Dsg1) antibody compared with patients with PV (P =0.047).However,there were no significant differences in the levels of anti-Dsg1 and anti-Dsg3 antibodies between PV patients who had acantholysis in the middle-to-upper epidermis and those in the upper basal layer.Conclusion Histopathological examinations of PV and PF lesions show that acantholysis can occur in the middle-to-upper epidermis,as well as in the middle-to-lower epidermis,and locations of acantholysis may be associated with levels of anti-Dsg1 and anti-Dsg3 antibodies.

Objective To explore the value of urinary albumin/creatinine ratio (ACR) in coronary heart disease(CHD) compli-cating thyroid dysfunction and its correlation with the various detection indicators of thyroid function.Methods The results of uri-nary ACR and thyroid function detection in 863 patients with CHD in our hospital from November 2015 to August 2016 were col-lected.According to the standards of American National Kidney Foundation (NKF) and Food and Drug Administration(FDA) ,863 CHD patients were divided into the group A (ACR≤10.0 mg/g · Cr) ,B(10.0 mg/g · Cr< ACR≤30.0 mg/g · Cr) ,C (30.0 mg/g · Cr300 mg/g · Cr).According to the reference values of thyroid functional indica-tors ,these patients were divided into the thyroid function normal and abnormal groups.Results The ACR level was negatively cor-related with the FT3 level in 863 CHD patients(r= -0.297 ,P<0.05) ,and was positively related with the FT 4 level(r=0.172 , P<0.05).The abnormal rate of thyroid function had significantly statistical difference among four groups (χ2 =33.533 ,P<0.05). The abnormal rate of thyroid function had significantly statistical difference between the patients with ACR >10.0 mg/g · Cr and the patients with ACR ≤ 10.0 mg/g · Cr ,the difference was statistically significant (P< 0.05) ,its risk degree was 2.083. Conclusion ACR is likely to be a risk factor for CHD patients complicated with thyroid dysfunction.It is necessary to monitor the thyroid function when ACR level >10.0 mg/g · Cr in CHD patients.

Objective To evaluate the regulatory effect of hydroxychloroquine on in vitro differentiation of regulatory T cells(Treg). Methods Fifteen milliliters of peripheral blood were obtained from a healthy human subject followed by isolation of monouclear cells and sorting of naive T cells. Then, the naive T cells were classified into two groups:control group cultured with the presence of tumor growth factor(TGF)?βand interleukin(IL)?2, experimental group cultured with the presence of 20μmol/L hydroxychloroquine and inducing factors. The number and percentage of CD4+Foxp3+Treg cells were determined by flow cytometry at days 0, 6, and 12. Results The number of Treg cells at days 0, 6, and 12 were(2 ± 0.4)× 104,(13.2 ± 5.2)× 104, and(143.5 ± 35.9)× 104 respectively in the control group, (2 ± 0.5)× 104,(2.4 ± 0.4)× 104, and(5.6 ± 3.5)× 104 respectively in the experimental group. There was a significant difference in the number of CD4+Foxp3+ Treg cells between the two groups at days 6 and 12 (t = 3.78 and 6.16, respectively, both P < 0.05). At day 12, the percentage of CD4 +Foxp3 + Treg cells was 79.7% ± 18.1% in the experimental group, compared to 16%± 13%in the control group(t=11.77, P<0.01). Conclusion Hydroxychloro?quine could inhibit the differentiation and proliferation of CD4+Foxp3+Treg cells in vitro.

Objective To optimize and establish the extractive technology of taxol from Taxus baccatal L.. Methods HPLC was applied to determine the content of taxol. The orthogonal test was adopted to examine the effects of the 4 factors consisting of ethanol concentration,ethanol volume,extractive times and extractive time. Results Ethanol concentration and ethanol volume were essential factors influencing the extraction of taxol. High concentration and amount of ethanol facilitated the extraction. Extractive times and extractive time were also important. By single-factorial effect,2 h and 2 times in each extraction of taxol were the best. Conclusion The optimum extractive condition is T9 (A3B3C2D1) and T8 (A3B2C1D3).

Objective To investigate the value of bone morphogenetic protein-7 (BMP-7) and the kidney shear wave velocity (SWV) in the diagnosis and treatment of diabetic nephropathy.Methods 150 cases of patients with diabetic mellitus were divided into three groups according to 24h urinary albumin excretion (Ualb):normalbuminuria (NA) group,microalbuminuria (MA) group and clinical proteinuria (CA) group;50 healthy subjects were selected as the control group (NC).The levels of blood BMP-7 were detected and the virtual touch tissue quantification (VTQ) was used to detect renal SWV in four groups.Both the BMP-7 level and the renal parenchyma SWV were compared among four groups.Then correlation analysis was made between blood of BMP-7 level and renal SWV in diabetic nephropathy groups.Results Blood BMP-7 level:the lowest was CA group,the middle one was MA group,and the highest was NA group (P ＜0.05),the NC group and the NA group had no significant difference (P ＞0.05); SWV measured values:CA group was the highest,MA group was middle one and the NA group was the lowest (P ＜ 0.05),the difference between NC group and the NA group was not significant statistically (P ＞0.05).The blood levels of BMP-7 and renal SWV had a significant negative correlation (r =-0.612,P ＜0.05).Conclusions The blood BMP-7 levels downregulate with the progressing of diabetic nephropathy,but the renal SWV increases with the diabetic nephropathy deterioration,and may indirectly reflects blood BMP-7 levels.Both of them have important application values in the diagnosis or treatment of diabetic nephropathy.

Objective To evaluate the effect of intraperitoneal water soluble lipopolymer (WSLP)/ N-methyl-D-aspartate receptor subunit 2B (NR2B) siRNA compound on the neuropathic pain (NP) in rats.Methods Eighty-four healthy male Sprague-Dawley rats,aged 6 weeks,weighing 180-220 g,were randomly divided into 7 groups (n =12 each) using a random number table:control group (group C),sham operation group (group S),NP group,WSLP/NR2B siRNA group (siWSLP group),WSLP/negative control siRNA group (ncWSLP group),PEI/NR2B siRNA group (PEI group) and WSLP group (WSLP group).NP was produced by ligation of the left L5 spinal nerve.In group S,the left L5 spinal nerve was only exposed,but not ligated.In group C,the rats underwent no treatment.Groups siWSLP,ncWSLP,PEI and WSLP received single intraperitoneal injection of WSLP/NR2B siRNA,WSLP/negative control siRNA,PEI/NR2B siRNA and WSLP compound 2 ml,respectively,at 10 days after NP.At 1 day before operation,7 days after operation,and 3,7,14 and 21 days after intraperitoneal injection,6 rats in each group were chosen randomly to measure mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL).At 3 days after intraperitoneal injection,the left 6 rats in each group were sacrificed and the spinal cord was removed for detection of NP2B mRNA expression (using PCR) and NR2B expression (by Western blot).Results Compared with group C,MWT was significantly decreased,and TWL was shortened on 7 days after operation and 3,7 and 21 days after intraperitoneal injection,and the expression of NR2B mRNA and protein was down-regulated on 3 days after administration in the other groups.Compared with group NP,MWT was significantly increased,and TWL was prolonged on day 3 and 7 after intraperitoneal injection,and the expression of NR2B mRNA and protein was down-regulated on day 3 after administration in siWSLP group.Conclusion Intraperitoneal WSLP/NR2B siRNA compound can effectively relieve the NP in rats.

Objective:To develop an HPLC with gradient elution method for the determination of loteprednol etabonate and the re-lated substances in loteprednol etabonate and tobramycin compound ophthalmic suspension. Methods:HPLC with gradient elution was performed with Inertsil ph phenyl column (250 mm × 4. 6 mm, 5 μm). The mobile phase A and B was 0. 25% acetic acid solution-acetonitrile (80 ∶20) and acetonitrile, respectively. The flow rate was 2. 0 ml·min-1 and the detection wavelength was 244 nm. The column temperature was 30 ℃ and the injection volume was 20 μl. Results:The main ingredient and the related substances could be well separated. Loteprednol etabonate had a good linear relationship within the range of 0. 001-1. 02 mg·ml-1(r=0. 999 9), and the average recovery was 99. 9%(RSD=0. 9, n=9). Conclusion:The assay method is sensitive, accurate and convenient with good res-olution. It can be applied to control the quality of loteprednol etabonate and tobramycin compound ophthalmic suspension.

Objective:To study the effects of PRF and recombinant hPDGF-AB,TGF-β1 and VEGF on the proliferation and adhe-sion of rat adipose tissue-derived stem cells(ADSCs)in vitro.Methods:ADSCs were cultured with PRF membrane and various do-ses of PDGF-AB,TGF-β1 and VEGF,cell adhesion was examined by adhesion assay after 2 culture,cell proliferation was examined by CCK-8 kit after 1 -7 d culture.Results:Cell adhesion assay showed that the adhesive numbers of rat ADSCs in PRF group were significantly higher than those in the negative group(P <0.05).The adhesive numbers of the ADSCs treated by PDGF-AB showed no significantly difference among different concentration groups(P >0.05).The adhesive numbers of the ADSCs treated by VEGF or TGF-β1 at different concentrations showed significant difference(P <0.05).CCK-8 kit assay showed that at different time points, the A values of ADSCs in PRF group were significantly higher than those of the negative control group(P <0.05).The A values of ADSCs in VEGF or PDGF-AB groups at different concentrations showed significant difference(P <0.05).The A values of rat AD-SCs in TGF-β1 group at different concentrations were lower than those in the negative control group(P <0.05).Conclusion:PRF as a combination of growth factors may stimulate the proliferation and adhesion of rat ADSCs in vitro.PDGF-AB and VEGF may stim-ulate the proliferation of rat ADSCs.TGF-β1 and VEGF may stimulate the adhesion of rat ADSCs in a dose-response manner to some degree.