Objective: To modify EBP(endotoxin binding peptide), clone and express the mutate of EBP gene and gain purified mEBP.Method: mEBPgene was cloned by PCR site-directed mutagenesis. PinpointXa-3/mEBP expression vector was designed to express human mEBP as a fusion protein in BL21 (DE3) pLysS. Digested engineering bacteria by lysozyme and collected inclusion bodies.Fusion protein was purified by Pinpoint TM Xa purification system and cleaved by factorXa,mEBP was purified by RP-HPLC. Results: Mutations at residues 5 and 18(Gln→Lys) was obtained by PCR site-directed mutagenesis, expressed and purified mEBP successfully.Conclusions: Obtaining of purified mEBP lay a foundation for its biological activity research.

Objective To explore the relationship between infant feeding practices and infants growth,especially the weight gain for length in the first 6 months of life. Methods Two-hundred healthy full-term singlet normal birth weight 5 - 6 months old infants and their main care givers were recruited in Kongjiang community health service center in Shanghai. The questionnaires included infants feeding pattern, feeding environment and care givers feeding behaviors, and were completed on-site by investigators. The birth weight was obtained. The weight and length of infants at 6 months were measured. Results There were 70/200 (35.0%) infants overweight (BMI for age Z score>+1) at 6 months. There were more overweight boys than girls (40.0% vs. 30.9%, x2 = 1.798, P = 0.180). Compared with normal weight infants, the overweight infants had same birth weight (3.30 ± 0.35 kg vs. 3.35 ± 0.32 kg, t =1.010, P = 0.314) and same length at 6 months (67.64 ± 2.10 cm vs. 67.91 ± 1.97 cm, t=- 0.896, P = 0.371). However, the overweight infants gained much more weight for length in the first six months of life. At 6 months, the weight and BMI of overweight infants was significantly higher than that of normal weight infants (9.16 ± 0.67 vs. 7.94 ±0.64, t = 12.324, P < 0.01;19.86 ± 1.24 vs. 17.41 ± 0.80, t = 14.953, P < 0.01). The rates of exclusive breastfeeding for at least four months, continued breastfeeding at six months, complementary food consistency, and complementary feeding frequency, were not significantly differenct between overweight and normal weight infants (x2 =0.723 - 2.701, P > 0.05). The grandparents played an important role in infants feeding in Shanghai. There were 39.0% infants fed only by grandparents, and 23.0% infants fed jointly by grandparents and parents (x2 = 0.175, P > 0.05).The care givers' educational level and knowledge of feeding skill were similar in overweight and normal weight infants (x2 = 0.446, t = 0.949, P > 0.05). However, the overweight infants were fed more quickly than normal weight infants (Z = 2.753, P < 0.01), and there was less language communication between care givers and overweight infants during feeding (Z = 2.932, P < 0.01). In overweight infants, the overfeeding behaviors of care givers were positively correlated with BAZ (r = 0.281, P < 0.05). Conclusions Overweight in early life is associated with some particular infants feeding behaviors of care givers in Shanghai. The long term effect of more rapid weight gain for length in the first 6 months of life on the risk of childhood obesity warranted to be further followed and researched.

OBJECTIVETo evaluate the clinical efficiency and safety of two-micron laser resection of the prostate-tangerine technique (TmLRP-TT) for the treatment of large-volume ( > 70 ml) prostate in patients with benign prostatic hyperplasia (BPH).

METHODSThis retrospective analysis included 80 BPH patients with the prostatic volume larger than 70 ml, all treated by TmLRP-TT. We comparatively analyzed the levels of hemoglobin and serum sodium before and after surgery, recorded intra- and post-operative com- plications, and followed up the patients at 6 and 12 months after operation for International Prostate Symptom Score (IPSS), quality of life (QOL), maximum flow rate (Qmax), and postvoid residual urine volume (PVR).

RESULTSAll the operations were successfully completed. The mean hemoglobin decreased (0.68 +/- 0.43) g/dl intraoperatively, but no apparent reduction was observed in serum sodium. Lower urinary tract symptoms were relieved significantly in all the cases. At 12 months after surgery, IPSS was decreased by 73.89% as compared with the baseline (20.03 +/- 6.9 vs 5.23 +/- 3.59), QOL by 64.55% (4.09 +/- 1.19 vs 1.45 +/- 1.36), and PVR by 79.30% (97.31 +/- 57.90 vs 20.14 +/- 24.20 ml), while Qmax increased by 140.42% ([8.04 +/- 3.62] vs [19.33 +/- 3.28] ml/s). The incidence of complications was low either intraoperatively or during the 12 months after operation.

CONCLUSIONTmLRP-TT is a safe and effective surgical endoscopic approach to the treatment of large-volume prostate in BPH patients.

Aged ; Follow-Up Studies ; Humans ; Laser Therapy ; methods ; Male ; Middle Aged ; Prostatic Hyperplasia ; surgery ; Retrospective Studies ; Transurethral Resection of Prostate ; methods ; Treatment Outcome

This study is to report the establishment of an UPLC-MS/MS method for the determination of plasma concentration of UA carried in self-microemulsifying drug delivery system (SMEDDS) and its pharmacokinetics in rats. It was used for determination and analysis when serum with internal standard was extracted from C18 solid-phase column. Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for separation. The mobile phase was acetonitrile -0.1% ammonia with gradient elution at the flow rate of 0.2 mL x min(-1). The column temperature was 40 degrees C and the detection wave length was 210 nm. It was detected by negative ion using electrospray ionization source (ESI) and scanned by multiple reaction ion monitoring (MRM) mode. The liner relationship of UA was very good in the range of 1.19-3 815.00 ng x mL(-1) (r = 0.999 0). Recovery rate of different concentrations were 87.42%-89.95%. The precision of inter-day and intra-day were less than 11%. The method developed in our study was proved to be sensitive, rapid and simple. It is suitable for the pharmacokinetic study of UA-SMEDDS in rats.
Animals ; Chromatography, High Pressure Liquid ; Drug Delivery Systems ; Emulsions ; chemistry ; Rats ; Tandem Mass Spectrometry ; Triterpenes ; blood ; pharmacokinetics

The change of yield and contents. of active compositions were studied while the fibrous roots were decayed naturally. HPLC method was used to detect the contents of active composition. The results show that fibrousroots could decrease the production of plant by 38.60% (20 g) and 30.99% (40 g), respectively. Treatment 1 could increase the contents of dihydrotanshinone and cryptotanshinone of Salvia miltiorrhiza f. alba by 26.08% and 22.64%, respectively. Compared with the comparison, treatment 2 decreased the contents of ihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A) of S. miltiorrhiza f. alba by 60.87%, 79.24%, 84.61% and 88.99%, respectively. Meanwhile, the total contents of the liposoluble constituents reduced by 86.27%. The different concentration of fibrousroots could increase the content of salvianolic acid B by 4.98% (20 g) and 23.64% (40 g), respectively. Meanwhile, the content of rosemary acid was increased by 4.98% (20 g) and 23.64% (40 g), respectively. The content of water-soluble constituents positively correlated to the mount of added fibrousroots, and the change was significantly. The result indicted that the decay of fibrousroots has a significant impact on the growth and the content of the active composition of S. miltiorrhiza f. alba under the condition of continuous cropping. Fibrousroots could decrease the content of biomass and liposoluble constituents significantly, which maybe one of the main factors to S. miltiorrhiza f. alba continuous cropping obstacle formation.
Benzofurans ; analysis ; Biomass ; Chromatography, High Pressure Liquid ; Diterpenes, Abietane ; analysis ; Plant Roots ; metabolism ; Salvia miltiorrhiza ; chemistry ; growth & development

Biological characteristic of Salvia miltiorrhiza f. alba in field was studied. HPLC method was used to determine the lipophilic constituents (dihydrotanshinone, cryptotanshinone, tanshinone, tanshinone II (A) and miltione) and hydrophilic constituents (salvianolic acid, rosemarinic acid). The results showed that the fresh weight of S. miltiorrhiza f. alba which cropped for 2 years was decreased by 80.47%, while dry weight decreased by 79.42%. The normal diameter of the root was 0.3-0.5 cm, however, the diameter was 0.2-0.4 cm after 2 years, it was said that the decrease of the root diameter was the main reason for the decrease of the yield. The average contents of dihydrotanshinone, cryptotanshinone, tanshinone, tanshinone II (A), miltione, salvianolic acid and rosemarinic acid were decreased by 35.26%, 32.26%, 19.35%, 3.39%, 64.40%, 66.93% in plant which continuously cropped for 2 years, respectively. The yield and active constituents were mostly effected in the plant of S. miltiorrhiza f. alba, which continuously cropped for 2 years.
Drugs, Chinese Herbal ; analysis ; Linear Models ; Quality Control ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry ; growth & development ; Time Factors

OBJECTIVETo observe the effect of overdose fluoride on the proliferation of rat's incisor ameloblast.

METHODS20 Wistar rats were divided randomly into 2 groups: Group I (Control); Group II 50 mg/L F(-) were given. After 8 weeks treatment, the AgNORs stain and TUNEL technique were applied to analyze the effect of fluoride on the proliferation and apoptosis of ameloblasts.

RESULTSThe imagination analysis results showed that proliferation of pre-secretion ameloblasts were inhibited in group II as compared with the control group (P < 0.001). There was significant increase of apoptosis with the trend of migration toward secretion stage.

CONCLUSIONThe mechanism of fluorosis mottled enamel may be the effect of overdose fluoride with inhibits proliferation and induces apoptosis of ameloblasts resulting in dysfunction of secretion or absorption of enamel matrix proteins.

Ameloblasts ; drug effects ; pathology ; Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Dental Enamel ; pathology ; Female ; Fluorides ; adverse effects ; Fluorosis, Dental ; pathology ; Incisor ; Male ; Random Allocation ; Rats ; Rats, Wistar

Objective To obtain the full length (FL ) and C‐terminal fragment of polymorphic membrane protein I (PmpI) of Chlamydia trachomatis serovar D ,and to study the immunogenicity of these proteins .Methods The target genes of PmpI‐FL and PmpI‐C were amplified by polymerase chain reaction (PCR) and inserted into the prokaryotic plasmid vector pGEX‐6P‐1 .The recombinant plasmids pGEX‐6P‐1/PmpI‐FL and pGEX‐6P‐1/PmpI‐C were separately transformed into Escherichia .coli ( E . coli) DH5αand were identified by enzyme digestion ,sequencing and PCR .After the identification ,the recombinant plasmids were separately transformed into E .coli BL21 and induced to express the proteins . The expected proteins were identified by Coomassie brilliant blue staining and Western blot ,then purified by glutathione S‐transferase (GST) MagBeads .The purified proteins were then injected into BALB/c mice to prepare the polyclonal antibodies against PmpI‐FL or PmpI‐C .Enzyme‐linked immune sorbent assay (ELISA) was used for the quantitative detection of the specific antibody .Results The lengths of cloned target genes PmpI‐FL and PmpI‐C were 2 659 bp and 1 195 bp ,respectively ,and the sequences were consistent with those of Chlamydia trachomatis serovar D in GenBank .The molecular masses of target proteins were 122 000 and 69 000 ,respectively ,which were confirmed by Coomassie brilliant blue staining and Western blot and then purified .The titers of the antibodies (anti‐PmpI‐FL and anti‐PmpI‐C) in sera of immunized mice detected by ELISA were 1∶12 800 and 1∶6 400 ,respectively .Conclusion The PmpI‐FL‐GST and PmpI‐C‐GST fusion proteins with high immunogenicity are successfully expressed and purified , which lays the foundation for further study .

Objective To use Excel worksheet function to achieve statistical analysis of the fourfold table qualitative data,and to improve knowledge of the.majority of researchers on the statistical capabilities of Excel.Methods The result of actual research published in Chinese Journal of Endemiology was the data source,and statistical analysis results of fourfold table data from Excel and from SAS software were contrasted.Results The worksheet function of Excel could realize the analysis fourfold table data chi-square test,obtained an accurate P values,but not just P ＞ 0.05 or P ＜ 0.05.The results of chi-square test of Excel were the same as the results from SAS software,but Excel could not obtain probability value of fisher exact probability.Conclusion As a dependent statistical analysis software,Excel is an easy to learn,easy to use,efficient and a good helper.In the absence of professional statistical software,Excel can be used to achieve data statistical analysis.

OBJECTIVETo evaluate the clinical efficacy and efficacy sustainable time of acupuncture in knee osteoarthritis (KOA).

METHODSThe non-randomized concurrent control trial was adopted. One hundred and ninety-three cases of KOA were divided into an immediate acupuncture group (group A, 97 cases) and a delayed acupunc-weeks at the end of treatment. In group B, the same acupuncture therapy was applied after waiting 4 weeks. The acupoints in the two groups were Liangqiu (ST 34), Dubi (ST 35), Zusanli (ST 36), Yanglingquan (GB 34), Yinlingquan (SP 9), Xuehai (SP 10), Xiyan (EX-LE 4), Xiyangguan (GB 33). WOMAC (Western Ontario and McMasters Universities Osteoarthritis) was used for the assessment of the primary index and VAS (visual analogue scale) was for the secondary index. The evaluation was accomplished by the patients at the beginning of trial, on the 4th and 8th weeks. In each group, 72 patients finished the trial and the data of the lost cases were included in the final data analysis.

RESULTSIn the 4th week of trial, WOMAC score was (25. 8+/-22.0) in group A difference (P<0. 001). VAS scorewas (31. 8+/-24. and was (43.8+/-22.2) in group B, indicating the significant 6) in group A and was (56. 6 +/-25. 8) in group B, indicating very significant difference (P<0. 001). In the 8th week, the efficacy was reduced slightly in the follow-up of group A, but it was improved apparently as compared Acupuncture relieves joint pain and improves joint function obviously.by th patiĩeffr,a Mtaetfti-?an tf ri-with that before treatment.

CONCLUSIONAcupuncture relieves joint pain and improves joint function obviously.The effect of acupuncture is still sustainable in 4 weeks after terminating the treatment.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Arthralgia ; therapy ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; therapy ; Treatment Outcome