BACKGROUND:Astragaloside Ⅳ is a major component of Huangqi,promoting proliferation and differentiation of bone marrow mesenchymal stem cells;however,the mechanism has been less reported yet.OBJECTIVE:To explore the effect of Astragaloside Ⅳ on expression of multiple hematopoietic growth factors in bone marrow mesenchymal stem cells.METHODS:Bone marrow mesenchymal stem cells were isolated from adult Wistar rats by using the method of adhesive culture and clone,and they were then plated on 96-well plate and separately incubated with 100 uL Astragaloside Ⅳ(25,50,100,200 g/L)for 72 hours.The cells in the control group were cultured with an equal volume of DMEM-LG culture liuquid.Indirect immunofluorescence was used to detect the biological activity,MTT method was used to evaluate the effect of Astragaloside Ⅳ on proliferation and differentiation of bone marrow mesenchymal stem cells,and RT-PCR method was used to measure the expression of hematopoietic growth factors in bone marrow mesenchymal stem cells.RESULTS AND CONCLUSION:The 3~(rd)-passage bone marrow mesenchymal stem cells highly expressed CD44 but lowly expressed CD45.As compared with control group,Astragaloside Ⅳ promoted proliferation of bone marrow mesenchymal stem cells in a time/dosage-dependent manner,in particular,the 200 g/L Astragaloside IV and 72-hour intervention(P＜ 0.05).SCF expression was significantly increased in the drug group compared with control group(P ＜ 0.01);however,TPO,GM-CSF,and TGF-β1 expressions were not changed significantly(P ＞ 0.05).Moreover,interleukin-3 expression was not found in the bone marrow mesenchymal stem cells.Astragaloside Ⅳ promoted in vitro proliferation of bone marrow mesenchymal stem cells,possibly involving in SCF secretion.

Objective To investigate clinical and pathological features of lupus erythematosus profundus (LEP)of the scalp in children.Methods A retrospective study was carried out on 5 children with LEP.The clinical and histopathological features,treatment and prognosis of LEP were analyzed.Results The 5 children with LEP included 2 boys and 3 girls with a median age at onset of 5 months (range,2-38 months) and a median clinical course of 15 months (range,4-72 months).Clinically,the patients presented with arc-shaped or circular purple atrophic plaques on the scalp complicated by alopecia.The occipitalia and tempora were the most commonly involved sites.Antinuclear antibodies (ANA) and extractable nuclear antigens (ENAs) were negative in all the patients.Main histopathological changes were hyaline degeneration of the fat,mucin deposition and local aggregation of lymphocytes in fat lobules.Of the 5 patients,2 were treated with oral prednisone (1.5-2 mg/kg/day),1 with oral hydroxychloroquine (5 mg/kg/day),1 with oral prednisone (1.5 mg/kg/day) combined with hydroxychloroquine (5 mg/kg/day),and another 1 with topical halometasone cream and 0.03％ tacrolimus ointment.Lesions were remissed after 2-3 months of treatment,and subsided with growth of new hairs after 6 months.No recurrence was observed during a 1.5-year follow-up.Conclusion Prednisone and hydroxychloroquine are markedly effective for LEP,and pediatric patients with LEP may be treated by topical highpotency glucocorticoids and calcineurin inhibitors.

To preliminarily study the pro⁃angiogenic activity of Echinococcus granulosus hydatid cysts against hu⁃ man umbilical vein endothelial cells in vitro and the transcriptional level of potential pro⁃angiogenic factors. Methods The hydatid cysts and protoscolex derived from experimentally infected mice were collected and cultured in vitro，then the human umbilical vein endothelial cells were stimulated by the supernatant and cyst fluid respectively，and the angiogenesis was observed and analyzed through a microscope and the angiogenesis mode of the software NIH Image J. Meanwhile，the mouse homologous proteins of matrix metalloproteinase⁃9（MMP⁃9）and high mobility group box B1（HMGB1）were identified in E. granulosus genome through sequence alignment，and their transcriptional levels in the cyst wall and protoscolex were analyzed. Results The culture supernatant of hydatid cysts significantly promoted human umbilical vein endothelial cells into tubes（F = 73.03，P < 0.001），the transcriptions of MMP⁃9 and HMGB1 were detected in the cyst wall and protoscolex，and the transcriptional level of MMP⁃9 was higher in protoscolex（t = -11.65，P < 0.001），while the level of HMGB1 was higher in hydatid cysts（t = 6.43，P = 0.003）. Conclusion Some parasite⁃derived pro⁃angiogenic molecules may exist in the supernatant of E. granulosus hydatid cysts，while further researches are required into their exact mechanisms.

Objective To study the structural features and characteristics of a novel gene Schistosoma japonicum 79(Sj79), and observe its effect of RNA interference(RNAi),so as to provide the experimental basis for its further function study and mechanism study of anti reproductive development of schistosome. Methods The gene structure and characteristics of Sj79 were analyzed by bioinformatics methods. Then the expressions of Sj79 messenger RNA(mRNA)during the different develop?mental stages of schistosome were analyzed and the effects of RNAi silencing were observed by the soaking method. The tran?scriptional levels of Sj79 after RNAi were detected by real time PCR. Results The open reading frame of Sj79 contained 696 base pairs with an exon structure. The gene had obvious stage specificity,and its transcriptional level in mature female worms was the highest. After soaking for 3 d,the Sj79 mRNA level[(41.0 ± 12.3)%]in the siRNA?1 group with low dosage(20 nmol/L) was lower than that in the siRNA?NC group[(103.2 ± 14.4)%],the difference was statistically significant(t=3.28,P<0.05). When with high dosage(200 nmol/L ),both the Sj79 mRNA levels in the siRNA?1 group[(15.8 ± 10.9)%]and siRNA?2 group [(11.1 ± 8.8)%]were significantly lower than that in the siRNA?NC group[(100.1 ± 6.3)%](t=13.44,27.84,both P<0.01). After soaking for 7 d,only the Sj79 mRNA levels in the siRNA?1group[(43.4 ± 4.5)%]and siRNA?2 group[(62.5 ± 5.4)%]with low dosage were lower than that in the siRNA?NC group[(100.4 ± 5.2)%],and the differences had statistical sig?nificance(t=8.33,5.07,both P<0.01). Conclusion Through this study,we have improved the mRNA sequence and genom?ic information of Sj79 gene,and understood its structural features,as well as selected out two effect fragments siRNA?1and siR? NA?2 which will provide the basic evidences for the further study on egg laying interference of the female adult worm of schisto?some in vitro.

Objective To better understand the knowledge, attitudes, practice (KAP) and relative demand of precon?ception and prenatal care (PAPC) in medical staffs of obstetrics and gynecology department in Tianjin. Methods Using ran?dom, stratified and clustered sampling approach, we selected 382 obstetrics and gynecology personnels in Tianjin. A self-ad?ministered questionnaire on PAPC KAP was employed to survey their general characteristics, their awareness of PAPC knowledge, their attitudes to PAPC, their service capabilities of PAPC and their request of continuing education. Results The respondents were lacking of KAP in Calcium and Iron supplementation, alternative preconception examination and emerging prenatal examination but present good KAP in folic acid supplemen, necessary prenatal items and classic prenatal examination. Doctors′average knowledge score was higher than nurses′. The scores in primary health care institutions was significantly higher than that in tertiary and secondary medical institutions. Young and junior medical respondents showed higher awareness than older and senior ones. Most (91.21%) respondents believe that PAPC is necessary. But 86.54%of re?spondents are not confident enough to provide comprehensive PAPC services. There were 63.74%, 54.67%, 38.74% and 32.42%of the respondents who could provide special PAPC advisory of adverse pregnancy history, thyroid dysfunction, re?productive infections and diabetes respectively. The most urgent training contents include PAPC knowledge in presence of complications or risk factors, advances in prenatal diagnosis, nutrition during pregnancy and consulting skills. Conclusion It′s necessary to intervent obstetrics and gynecology personnels to improve their cognitive level to PAPC, as well as to amelio?rate relative attitudes and behavior so they can provide more scientific and comprehensive PAPC service.

Objective To study the inhibitory effect of antimicrobial peptide LL-37 on Candida albicans through its ability to promote the secretion of immune factors by vaginal epithelial cells. Methods (1) LL-37 prokaryotic expression vector pET-Duet/LL-37 was constructed and its expression was induced in Escherichia coli M15. The expressed LL-37 fusion protein was purified and identified by western blot. Antifungal activity of the purified protein was initially identified by Kirby-Bauer (K-B) method. (2) Purified LL-37 protein was added to human vaginal epithelial cells co-cultured with Candida, and inhibitory effect on Candida growth was determined by the glucose consumption method. Interferon γ (IFN-γ), interleukin 10 (IL-10) concentration and IFN-γ/IL-10 ratio were measured by ELISA at different time points. Results (1) LL-37 fusion protein was purified to 96% purity at a concentration of 433.92 μg/ml, and was shown to possess anti-fungal activity confirmed by the K-B method. (2) A Candida-vaginal epithelial cells co-culture system was successfully constructed. LL-37 recombinant protein inhibited the growth of Candida with absorbance values significantly higher in the treatment group compared to the control group at all measured time points (12-hour:3.008±0.003 versus 2.967±0.003, 24-hour:2.941±0.003 versus 2.601±0.003, 48-hour:2.893 ± 0.004 versus 2.409 ± 0.003; all P<0.01). Furthermore, the rate of decrease was also much slower compared to the control group. In both control and experimental groups, IFN-γ and IL-10 secretion levels were observed to rise at first peaking at 24 hours and subsequently decrease. For each time period, IFN-γconcentration in the experimental group was significantly higher at 24 hours compared to the control group [(104.00 ± 1.07) versus (85.17 ± 0.28) pg/ml, P<0.01]. In contrast, IL-10 concentrations were significantly lower than the control group at all time points (P<0.01). IFN-γ/IL-10 ratio was also observed to be significantly higher than the control group at all measured time points (P<0.01). Conclusions (1) Recombinant protein LL-37 could significantly inhibit the growth of Candida. (2) By influencing the secretion of immune factors such as IFN-γ, IL-10, etc, recombinant protein LL-37 is able to adjust vaginal epithelial cells local immunity, and enhance resistance to Candida infection.

Objective To investigate the relationship between the right-to-left shunt(RLS)detected with contrast-enhanced transcranial Doppler (c-TCD) and recurrent stroke in patients with cryptogenic stroke.Methods The consecutive patients w ith ischemic stroke w ere enrol ed. The patients w ith cryptogenic stroke w ere screened according to the TOAST criteria. They w ere divided into either a RLS positive group or a RLS negative group according to the c-TCD findings, and then they w ere fol ow ed up for a period of one year. They w ere also divided into a recurrent group and a non-recurrent group according to w hether they had recurrence or not. Results A total of 118 patients w ith cryptogenic ischemic stroke w ere enrol ed, including 46 in the RLS positive group, 72 in the RLS negative group, 10 in the recurrent group, and 108 in the non-recurrent group. There w ere no significant differences in demographic and baseline data betw een the RLS negative group and the RLS positive group. There w ere significant differences in RLS positive rate (7/10 vs.39/108; P=0.046) and proportion of patients with server RLS (2/10 vs.1/108; P=0.019) betw een the recurrent group and the non-recurrent group. Multivariate logistic regression analysis show ed that the positive RLS w as an independent predictor of recurrent stroke (odds ratio 4.896, 95% confidence interval 1.135-21.120;P=0.033). Conclusions The positive RLS may be an independent risk factor for the recurrence in patients w ith cryptogenic ischemic stroke.

OBJECTIVETo compare the oncology outcomes and anal function among laparoscopic partial, subtotal and total intersphincteric resection(ISR) for low rectal cancers.

METHODSFrom June 2011 to February 2016, a total of 79 consecutive patients with low rectal cancers underwent laparoscopic ISR with hand-sewn coloanal anastomosis at our department. According to the distal tumor margin, partial ISR (internal sphincter resection at the dentate line) was used to treat tumors with distance <1 cm from the anal sphincter (n=28), subtotal ISR was adopted for the tumors locating between the dentate line and intersphincteric groove (n=34), and total ISR (resection at the dentate line) was applied in the treatment of intra-anal tumors (n=17). Anal function was evaluated by a standardized gastrointestinal questionnaire, Wexner incontinence score and Kirwan's classification. Metaphase oncological results and postoperative anal function were compared among three groups, and.

RESULTSOther than the distance of tumor low margin to dentate line (P=0.000) and serum CEA level (P=0.040), no significant differences were noted in baseline data among 3 groups (all P>0.05). The median follow up was 21(8-61) months. The 3-year disease-free survival rates in laparoscopic partial, subtotal and total ISR groups were 91.1%, 88.9%, 88.2% (P=0.901) and the 3-year local relapse-free survival rates were 91.1%, 72.9%, 80.2%(P=0.658), whose all differences were not significant. Thirty-eight patients who did not receive neoadjuvant chemoradiotherapy and underwent ileostomy closure for at least 24 months completed the evaluation of anal function, including 14 cases in partial group, 15 cases in subtotal group and 9 cases in total group. Of 38 patients, 73.7%(28/38) was classified as good function (Wexner incontinence score ≤10) and no patient adopted a colostomy because of severe fecal incontinence(Kirwan classification=grade 5). Furthermore, there were no significant differences in Wexner incontinence score and Kirwan classification among 3 groups (all P>0.05). However, patients with chronic anastomotic stoma stenosis showed worse anal function than those without stenosis [Wexner incontinence score: 18(9-20) vs 6(0-18), P=0.000; Kirwan grading: 3(2-4) vs. 2(1-4), P=0.002].

CONCLUSIONSAs the ultimate sphincter-saving technique, laparoscopic ISR can result in better oncologic outcomes and better anal function for patients with low rectal cancers. The different procedures of ISR may not affect the efficacy, but chronic anastomotic stoma stenosis deteriorates incontinence status.

OBJECTIVETo summarize the perioperative and postoperative complications follow laparoscopic intersphincteric resection (LapISR) in the treatment of low rectal cancer and their management.

METHODSAn observational study was conducted in 73 consecutive patients who underwent LapISR for low rectal cancer between June 2011 and February 2016 in our hospital. The clinicopathological parameters, perioperative and postoperative complications, and clinical outcomes were collected from a prospectively maintained database. Perioperative and postoperative complications were defined as any complication occurring within or more than 3 months after the primary operation, respectively.

RESULTSForty-nine(67.1%) cases were male and 24(32.9%) were female with a median age of 61(25 to 79) years. The median distance from distal tumor margin to anal verge was 4.0(1.0 to 5.5) cm. The median operative time was 195 (120 to 360) min, median intra operative blood loss was 100 (20 to 300) ml, median number of harvested lymph nodes was 14(3 to 31) per case. All the patients underwent preventive terminal ileum loop stoma. No conversion or hospital mortality was presented. The R0 resection rate was 98.6% with totally negative distal resection margin. A total of 34 complication episodes were recorded in 21(28.8%) patients during perioperative period, and among which 20.6%(7/34) was grade III(-IIII( according to Dindo system. Anastomosis-associated morbidity (16.4%,12/73) was the most common after LapISR, including mucosa ischemia in 9 cases(12.3%), stricture in 7 cases (9.6%, 4 cases secondary to mucosa necrosis receiving anal dilation), grade A fistula in 3 cases (4.1%) receiving conservative treatment and necrosis in 1 case (1.4%) receiving permanent stoma. After a median follow up of 21(3 to 60) months, postoperative complications were recorded in 12 patients (16.4%) with 16 episodes, including anastomotic stenosis (8.2%), rectum segmental stricture (5.5%), ileus (2.7%), partial anastomotic dehiscence (1.4%), anastomotic fistula (1.4%), rectovaginal fistula (1.4%) and mucosal prolapse (1.4%). These patients received corresponding treatments, such as endoscopic transanal resection, anal dilation, enema, purgative, permanent stoma, etc. according to the lesions. Six patients (8.2%) required re-operation intervention due to postoperative complications.

CONCLUSIONAnastomosis-associated morbidity is the most common after LapISR in the treatment of low rectal cancer in perioperative and postoperative periods, which must be strictly managed with suitable methods.

Adult ; Aged ; Anal Canal ; surgery ; Anastomosis, Surgical ; adverse effects ; Blood Loss, Surgical ; statistics & numerical data ; Colectomy ; adverse effects ; Constriction, Pathologic ; etiology ; therapy ; Digestive System Surgical Procedures ; adverse effects ; Female ; Humans ; Ileostomy ; adverse effects ; Intestinal Mucosa ; pathology ; Ischemia ; etiology ; Laparoscopy ; adverse effects ; Lymph Node Excision ; statistics & numerical data ; Male ; Margins of Excision ; Middle Aged ; Necrosis ; etiology ; Operative Time ; Postoperative Complications ; etiology ; therapy ; Rectal Neoplasms ; complications ; surgery ; Rectovaginal Fistula ; etiology ; therapy ; Surgical Stomas ; Treatment Outcome

OBJECTIVECryptosporidium spp. are prevalent globally and sheep are an important zoonotic reservoir. Little data regarding the rates of Cryptosporidium infections in ovines in China are available. This study assessed the prevalence of Cryptosporidium spp. in pre-weaned ovines from Aba Tibetan and Qiang Autonomous Prefecture in the Sichuan province of China.

METHODSA total of 213 fecal samples were collected from pre-weaned ovines and were examined microscopically (following modified acid fast staining). In addition, 18S rRNA genetic sequences were amplified from fecal samples by nested PCR and phylogenetically analyzed.

RESULTSThe prevalence of Cryptosporidium in the collected samples was at 14.6% (31/213) and four isolates identified by PCR belonged to the Cryptosporidium cervine genotype (Cryptosporidium ubiquitum) demonstrating that this species was the primary sheep species found in sheep in China.

CONCLUSIONThe present study suggested that the high incidence of Cryptosporidium in sheep poses a significant public health threat and that surveillance practices must be established to prevent zoonotic disease of humans.

Animals ; China ; Cryptosporidium ; genetics ; isolation & purification ; Feces ; parasitology ; Oocysts ; microbiology ; Polymerase Chain Reaction ; Sheep ; Weaning