In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Simple and sensitive methods were developed for the determination of indapamide, perindopril and its active metabolite perindoprilat in human plasma or whole blood by hyphenated ultra-performance li-quid chromatography-mass spectrometry (UPLC-MS/MS). Indapamide-d3, perindopril-d4 and perindo-prilat-d4 were used as the internal standards. The separation was performed on a Thermo BDS Hypersil C18column (4.6 mm × 100 mm, 2.4 μm) for indapamide and perindopril simultaneously following a protein precipitation pretreatment of the biosamples. The separation of perindoprilat was achieved in-dependently on a phenomenex PFP column (4.6 mm × 150 mm, 5 μm). All the analytes were quantitated with positive electrospray ionization and multiple reactions monitoring mode. The assay exhibited a linear range of 1–250 ng/mL for indapamide, 0.4–100 ng/mL for perindopril and 0.2–20 ng/mL for peri-ndoprilat. The methods were fully validated to meet the requirements for bioassay in accuracy, precision, recovery, reproducibility, stabilities and matrix effects, and successfully applied to the pharmacokinetic study of perindopril tert-butylamine/indapamide compound tablets in Chinese healthy volunteers and the comparative pharmacokinetic study between plasma and whole blood.

Objective To observe the anti-tumor action of Chang'aishu and its influence on the expression of the Ki67 and PCNA. Methods Forty Balb/c mice with CT-26 colorectal cancer were randomly divided into four groups, including the control group (normal saline, 0.6 mL), the chemotherapy group (Xeloda 205.5 mg/kg), the high dose group (Chang'aishu 51.38 g/kg) and the low dose group (Chang'aishu 17.13 g/kg). Each group of mice was treated with intragastric administration every two days. After 15 days, the anti-tumor rate was calculated and the expression of Ki67 and PCNA were analyzed by immunohistochemistry. Results Compared with the control group, inhibitive rate of the chemotherapy group, the high dose group and the low dose group was 43.35%, 29.48% and 13.30%, respectively. Compared with the control group, the treatment group showed statistical significant difference in Ki67 and PCNA (P<0.05, P<0.01). Conclusion Chang'aishu had inhibition effect on the growth of colorectal cancer, which may be related to down-regulating the expression of Ki67 and PCNA.

Objective To evaluate the clinical effects of a single posterior translaminal or transforaminal debridement and internal fixation with bone graft for the treatment of lumbosacral regional spinal tuberculosis. Methods From January 2004 to October 2009, 32 patients with lumbosacral regional spinal tuberculosis were treated by a single posterior debridement and internal fixation with bone graft, including 21 males and 11 females with the mean age of 46 years (range, 32-65). The lesions located: 3 in L3,4, 19 in L4,5, 10 in L5S1. 20 cases were pedormed operation via translaminal approach, 12 via transforaminal approach. The modified Prolo scale, visual analogue scale (VAS), kyphosis angle and the vertebral body loss were measured before and after surgery, and in the final follow-up. Results All the patients were followed up for average 16 months (range, 12-24). The pre-, postoperative and final follow-up mean kyphosis angle were -24.3°,-34.8°, and -32.4° in the translaminal group, -25.4°, -35.2°, and 33.6° in the transforaminal group,respectively. According to the modified Prolo scale, the excellent and good result was 93.8%. VAS decreased significantly from 7.6 points to 2.9 after surgery. Conclusion Debridement and internal fixation with bone graft via a single posterior approach for the treatment of lumbosacral regional spinal tuberculosis was an easy and effective technique. The appropriate selection of indications, careful imaging evaluation and thorough surgical debridement played an important role in the treatment of lumbosacral regional spinal tuberculosis.

According to the 2010 Chinese pharmacopeia, salt processed and charcoal processed Eucommiae Cortex were pre- pared. HPLC-DAD analysis of the content of the bark and leaf of Eucommiae Cortex showed that the bark of Eucommiae Cortex mainly contained lignans such as pinoresinol glucose and iridoid including genipin, geniposide, geniposidic acid, while the leaf of Eucommiae Cortex consisted of flavonoids such as quercetin and phenolic compound such as chlorogenic acid. The content of pinoresinol diglucoside in the bark of Eucommiae Cortex was about 18 times more than that in the leaf of Eucommiae Cortex. The content of pinoresinol diglucoside in salted and charcoal processed Eucommiae Cortex decreased approximately by 30% and 85%, respectively. The content of genipin, geniposide and geniposidic acid in the bark of Eucommiae Cortex was about 3 times, 23 times, 28 times more than that in the leaf of Eucommiae Cortex. The content of genipin, geniposide and geniposidic acid in salted Eucommiae Cortex were reduced by 25%, 40% and 40%, respectively. The content of genipin, geniposide and geniposidic acid in charcoal processed Eucommiae Cortex were reduced by 98%, 70%, 70%, respectively. The content of caffeic acid in bark of Eucommiae Cortex was about 3 times more than that in the leaf of Eucommiae Cortex. The content of caffeic acid was decreased by about 50% in the salted Eucommiae Cortex. While the content of caffeic acid in charcoal processed Eucommiae Cortex was decreased approximately 75%; the content of chlorogenic acid in bark of Eucommiae Cortex was about 1/6 of that in the leaf of Eucommiae Cortex. The content of chlorogenic acid in salted and charcoal processed Eucommiae Cortex decreased by 40% and 75%, respectively; the content of quercetin in bark of Eucommiae Cortex was only 1/40 of that in the leaf of Eucommiae Cortex. The content of quercetin in salted and charcoal processed Eucommiae Cortex were reduced by 60% and 50%, respectively.
Drugs, Chinese Herbal ; chemistry ; Eucommiaceae ; chemistry ; Flavonoids ; chemistry ; Plant Bark ; chemistry ; Plant Leaves ; chemistry

Objective : To evaluate the effectiveness of interventions for human papillomavirus ( HPV ) vaccine hesitancy among female university students based on the precaution adoption process model (PAPM), so as to provide the evidence for improving the coverage of HPV vaccine in this population. Methods : HPV vaccine hesitant female students were selected using a cluster sampling method from Zhejiang Chinese Medical University, and randomly assigned to the PAPM group and control group. Students in the PAPM group received PAPM-based interventions for HPV vaccine hesitancy, while students in the control group were given routine interventions. The HPV-related knowledge, HPV vaccine-related knowledge and HPV vaccine hesitancy scores were collected from both groups prior to interventions ( T0 ), 0 ( T1 ), 1 ( T2 ) and 3 months post-interventions ( T3 ), and the effectiveness of interventions was evaluated using analysis of variance for repeated measures. Results : There were 147 students in the PAPM group and 141 students in the control group. In the PAPM group, 36.73% of the students majored in medical sciences, and 48.23% were freshmen; in the control group, 39.72% majored in medical sciences, and 50.35% were freshmen. The mean scores of HPV- and HPV vaccine-related knowledge were significantly greater in the PAPM group than in the control group, respectively ( T1, 5.29 vs. 4.91; T2, 4.27 vs. 4.22; T3, 4.22 vs. 4.04; P<0.05 ); however, no significant differences were detected in the HPV vaccine hesitancy scores between the two groups, respectively ( T1, 2.98 vs. 2.95; T2, 3.07 vs. 3.07; T3, 3.08 vs. 2.97; P>0.05 ). The mean scores of the confidence dimension ( T1, 3.37 vs. 3.23; T2, 3.48 vs. 3.40; T3, 3.38 vs. 3.25 ) and the dimension of influence by others ( T1, 3.44 vs. 3.33; T2, 3.42 vs. 3.37; T3, 3.46 vs. 3.27 ) were significantly greater in the PAPM group than in the control group (P<0.05), while the mean scores of the complacency dimension were significantly lower in the PAPM group than in the control group ( T1, 1.98 vs. 2.03; T2, 2.06 vs. 2.20; T3, 2.18 vs. 2.15; P<0.05 ). Conclusions PAPM-based interventions for HPV vaccine hesitancy may effectively improve the awareness of HPV and HPV vaccines, reduce complacency, and enhance the influence by others among female university students.

OBJECTIVETo compare the effect of transurethral resection of the prostate combined with endocrine therapy (TURP + ET) with that of αlA-blockers combined with ET ((αlA-b + ET) in the treatment of bladder outlet obstruction (BOO) in patients with advanced prostate cancer (PCa), and to investigate the safety of the TURP + ET for the treatment of PCa with BOO.

METHODSWe retrospectively analyzed 63 cases of PCa with BOO, 28 treated by αlA-b + ET and the other 35 by TURP + ET. We obtained the residual urine volume (RV), maximum urinary flow rate (Qmax), International Prostate Symptom Score (IPSS), and quality of life score (QoL) before and after treatment along with the overall survival rate of the patients, followed by comparison of the parameters between the two methods.

RESULTSAt 3 months after treatment, RV, IPSS, and QoL in the TURP + ET group were significantly decreased from (137.8 ± 27.6) ml, (22.3 ± 3.6), and (4.2 ± 0.8) to (29 ± 13.6) ml, (7.8 ± 2.1), and (1.6 ± 0.5) respectively (P < 0.05), while Qmax increased from (5.6 ± 2.1) ml/s to (17.6 ± 2.7) ml/s (P < 0.05); the former three parameters in the αlA-b + ET group decreased from (133.6 ± 24.9) ml, (21.5 ± 3.2), and (4.7 ± 1.1) to (42 ± 18.3) ml, (12.8 ± 2.6), and (2.5 ± 0.7) respectively (P < 0.05), while the latter one increased from (6.3 ± 2.4) ml/s to (11.7 ± 2.3) ml/s (P < 0.05), all with statistically significant differences between the two groups (P < 0.05). The overall survival rate of the TURP + ET group was not significantly different from that of the αlA-b + ET group (51.4% vs 46.4% , P > 0.05).

CONCLUSIONTURP + ET is preferable to αlA-b + ET for its advantage of relieving BOO symptoms in advanced PCa without affecting the overall survival rate of the patients.

Adrenergic alpha-1 Receptor Antagonists ; therapeutic use ; Antineoplastic Agents, Hormonal ; therapeutic use ; Combined Modality Therapy ; methods ; Humans ; Male ; Prostatic Neoplasms ; complications ; drug therapy ; pathology ; surgery ; Quality of Life ; Retrospective Studies ; Transurethral Resection of Prostate ; Treatment Outcome ; Urinary Bladder Neck Obstruction ; drug therapy ; etiology ; surgery

Objective To investigate the prognostic factors affecting the postoperative survival of patients with invasive bladder cancer,and to predict the survival time of the patients.Methods We retro- spectively analyzed the clinical and follow-up data of 178 patients with invasive bladder cancer treated by radical cystectomy and urinary diversion from 1991 to 2004.A multivariate analysis was performed in these patients by the Cox proportional hazard model.A prognostic index(PI)based on the Cox regression was con- structed.According to the individualized PI,the patients were classified into different hazard groups and the expected survival curve of each patient was calculated.Results Cox regression analysis showed that the factors which influenced the postoperative survival included tumor stage(RR=1.982,P=0.000),grade (RR=1.978,P =0.042),lymph node metastasis(RR=2.142,P=0.048),Tis(RR=6.177,P= 0.000),tumor shape(RR=0.416,P=0.003),number of tumors( RR=1.820,P=0.035),pathological type(RR=2.228,P=0.032),patient age(RR=0.672,P=0.025)and neoadjuvant chemotherapy (RR=0.257,P=0.016).Based on the percentile of PI,patients were classified into 3 prognostic groups; the median survival time of 3 groups were 42.5,22.5 and 7.0 months,respectively.There were significant differences between each 2 among the 3 groups(P＜0.01).Conclusions Neoadjuvant chemotherapy, tumor stage,grade,lymph node metastasis,Tis,shape and number of tumors,pathological type,patient age were important prognostic factors.PI value can be used to predict the prognosis of patients with invasive blad- der cancer.

Objective To evaluate the acute and delayed killing effect of high intensity focused ultrasound (HIFU) on Echinococcus granulosus(E. granulosus)protoscolices in vitro.Methods E. granulosus protoscolices were treated with different dosage of effective power(0,25,50,100,200,250 W)and time(5,10,20,30,40,50,60 s)of HIFU in vitro to obtain the dosage-effect curves.Then the survival pmtoscolices were incubated,and the mortality of each group was counted daily.The protoscolicidal effects were investigated by trypan blue exclusion assay.Results Compared with the untreated group,the Vitality of E.granulosus protoscolices significantly decreased immediately after treated by HIFU of different dosage(F=5201.59 vs 1865.65,P＜0.05),there were the interaction both different dosage and time(F=214.50,P＜0.05).The protoscolices were broken into pieces by HIFU of 250 W×40 s,whereas the growth of the surviving protoscolices after exposed to HIFU was obvious suppressed.Both the acute killing effect and the delayed inhibitory effect showed a dosage-dependant manner.The inhibitory effect increased along with the increased dosage of HIFU(P＜0.05).The inhibitory effect in 50 W×10 s group was stronger than 25 W×20 s group(P＜0.05).The mortality was increased in parallel with the increase of HIFU dosage.Conclusions HIFU show an effective immediately killing effect,as well as a growth-inhibiting effect on the E.granulosus protoscolices in vitro.

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley