OBJECTIVE:To determine the concentration of clindamycin in woman milk by RP-HPLC.METHODS:The concentration of clindamycin(with tinidazole served as internal standard) was determined by liquid-liquid extraction-HPLC.The chromatographic column was Luna C18 and the mobile phase consisted of acetonitrile-tetrahydrofuran-phosphate buffer (pH 5)(30∶1∶70) at a flow rate of 0.8 mL?min-1.The temperature of the column was kept at 40 ℃ and the detection wavelength was set at 205 nm.RESULTS:The linear range of clindamycin was 0.529～10.580 ?g?mL-1(r=0.996 7) with a lowest detectable limit of 0.2 ?g?mL-1.The average methodological recovery rate of clindamycin was 98.84%;both the inter-day RSD and intra-day RSD of clindamycin(at low,medium and high concentrations) were all less than 5%.CONCLUSION:The method is simple,rapid,sensitive,accurate and reproducible,and it is applicable for the determination of clindamycin concentration in woman milk.

Background and purpose: AMP-activated protein kinase (AMPK) plays an important role in the regulation of cell metabolism and energy balance and is associated with cell proliferation, survival and multiple signaling pathways. Recent reports found that AMPK is involved in tumor suppression and drug resistance. The aim of this study was to explore the effect of AMPK on the anti-tumor effect of adriamycin and underlying mechanism in breast cancer MCF-7/adr cells. Methods:The anti-proliferative effects of adriamycin was detected by methyl thiazolyl tetrazolium (MTT) assay in MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKαcells. The cell morphology in each group was stained with the lfuorescent dye Hoechst 33528, and the effects on apoptosis induction were examined by lfow cytometry (FCM). The intracellular concentration of adriamycin was detected by lfuorescence assay. The resis-tance-and apoptosis-related proteins were analyzed by Western blot. Results:The growth of breast cancer MCF-7/adr cells was inhibited by adriamycin in a dose-and time-dependent manner. The IC50 values at 24 and 48 h were (36.8±2.1) and (28.8±1.3) μg/mL, respectively. AMPKαover-expression enhanced the cytotoxic effect of adriamycin in MCF-7/adr-AMPKαcells in a dose-and time-dependent manner. Its IC50 values at 24 and 48 h were (16.0±0.7) and (4.2±0.2) μg/mL, respectively. Fluorescent morphological assay showed that AMPKαoverexpression contributed to adriamycin induced apoptosis in MCF-7/adr-AMPKαcells. After treatment with 1.0 μg/mL adriamycin for 48 h, the apoptosis rates of MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKα cells were (12.0±1.4)%, (12.7±1.6)% and (32.0±4.2)%, respectively, indicating that overexpression of AMPKα enhanced the adriamycin-induced apoptosis in MCF-7/adr cells. Fluorescence microplate assay showed that over expression of AMPKαsigniifcantly increased the intracellular accumulation of adriamycin, in a concentration dependent manner. Western blot analysis showed that, compared with MCF-7/adr and MCF-7/adr-vector cells, the expressions of Bax, caspase-3 and cleaved PARP proteins were increased. Meanwhile, Bcl-2 and P-gp protein expressions were decreased in MCF-7/adr-AMPKαcells. Furthermore, the release of cytochrome c from mitochondria into the cytosol was also observed in MCF-7/adr-AMPKαcells. Conclusion:AMP-Kαoverexpression can enhance the chemosensitivity of breast cancer MCF-7/adr cells to adriamycin through inhibiting the drug effux transporter and regulating the expression of apoptosis-related proteins.

Objective To investigate the application value for predicting human cytomegalovirus(HCMV) infection with viral load in urinary epithehal cell (EC).Methods Peripheral blood and urine specimens from 82 infants with HCMV latent infection and 84 infants with HCMV active infection were collected respectively.Plasma HCMV DNA load and the levels of HCMV lgM/IgG antibody were detected by real-time fluorescence quantitative polymernse chain reaction (FQ-PCR) and chemiluminescence immunsassay.HCMV pp65 antigen in peripheral blood polymorphonuclear leukocytes (PMNLs) was detected by indirect immunofluorescence assay.The urinary EC count and HCMV DNA load were detected by UF-100 automated urine sediment analyzer and FQ-PCR,respectively.HCMV DNA load in urinary EC was calculated accordingly.At the same time,the sensitivity and specificity for diagnosis of active HCMV infection were evaluated by receiver operating characteristic curve (ROC).Results The positivity of HCMV DNA in urinary EC was 94.58% (157/166),which was the highest among the urinary EC from 166 cases of HCMV infection.HCMV DNA load ranged from 5.67×102to 1.31×107 copies/103 EC There was no significantly statistical difference among urine specimens from different periods of time(P>0.05).HCMV DNA load in active infection group [5.13±0.99(copies/103EC,lg)]is significantly higher than that in latent infection group [3.92±0.82 (copies/103 EC,lg),t = 8.52,P < 0.01];According to ROC curve analysis,its sensitivity and specificity were 71.4% and 75.2% respectively when cut-off value was 4.55.The virus load was significantly decreased in urinary EC in post-treatment infants as compared with pre-trestment(t=5.44,P<0.01).Conclusion Detection of HCMV DNA load in the urinary EC is a cost-effective method and can be used to predict HCMV active infection in infants and monitor treatment of HCMV infection.

Objective To investigate the diagnosis,surgical therapy of congenital pyloric atresia in neonates.Method Six congenital pyloric atresia neonates in Children's Hospital of Nanjing Medical University were admitted,including 4 cases of complete atresia with pyloric diaphragm,1 case of incomplete atrsia with a foraminula in the pyloric diaphragm and 1 case of pyloric atresia with solid segment.Three cases were associated with epidermolysis bullosa,multiple intestinal atresia and annular pancreas respectively.Results The main presenting symptoms were nonbilious vomiting,and 5 cases of abdominal X-ray plain film showed a large single gastric air-bubble and no gas distally.Ultrasonography and upper gastrointestinal radiography showed complete gastric outlet obstruction,and in 1 case postbulbar obstruction.Neonates with pyloric diaphragm underwent diaphragm excision and pyloroplasty,and that with solid segment did an extended pyloroplasty.The one complicating intestinal atresia was abandened surgery.Five cases were followed up,and doing well with complete recovery.Conclusion Abdominal X-ray plain film,Doppler ultrasonography and upper gastrointestinal radiography help establish the diagnosis of neonatal congenital pyloric atresia.Surgery is the therapy of choice and the prognosis is very good.

Total 3 483 students in 4 to 9 grade from primary and middle school in Xiling District of Yichang city were selected by stratified cluster random sampling method between September and October 2016 to participate in screening for abnormal angle of trunk rotation(ATR),including 1 797 boys and 1 686 girls aged 9-15 years.The ATR was measured with Scoliometer and the ATR >5°was defined as abnormal.The overall abnormal ATR rate was 7.60%(265/3 483), that in boys was 4.73%(85/1797)and in girls was 10.68%(180/1 686)(χ2=43.75,P<0.05).The abnormal ATR rate in age group 13-15 was higher than that in age group 9-12[10.62%(159/1 496)vs.5.33%(106/1 987),χ2=36.92,P<0.01].Students with low BMI had higher abnormal ATR rate than that with normal or higher BMI(χ2=30.00,P<0.01);and students with left handedness had lower abnormal ATR rate than that with right handedness or double handedness(χ2=6.59,P=0.04).The abnormal ATR rate was not associated with dietary habits or sports in the daily life of students.The results show that gender,age,BMI and handedness affect abnormal rate of ATR.

Objective: To evaluate the efficacy and safety of intravascular ultrasound guidance drug-coated balloon (DCB) in percutaneous coronary intervention (PCI) of situ coronary artery lesions in patients with acute coronary syndrome (ACS), and its effect on thrombus precursor protein (TpP). Methods: Seventy-eight patients with ACS in Central Hospital of Changchun City from January 2015 to January 2019 were selected, including 46 cases with unstable angina pectoris and 32 cases with acute non-ST-segment elevation myocardial infarction. The patients were divided into DCB group (38 cases) and drug-eluting stent (DES) group (40 cases) by random digits table method. Intravascular ultrasound was used to guide PCI in both groups, and DCB and DES were used respectively. Coronary angiography was performed immediately and 6 months after PCI in both groups. Minimum lumen diameter (MLD) was measured by QCA system, and the lumen loss (LLL) was calculated at 6 months after PCI. Plasma TpP before PCI, 1 and 6 months after PCI was measured by enzyme-linked immunosorbent assay (ELISA). Major adverse cardiovascular events (MACE) including cardiac death, myocardial infarction and target lesion revascularization (TLR) were followed up 1, 3, 6 and 12 months after PCI. Results: Immediately after PCI, there was no statistical difference in MLD between DCB group and DES group: (1.87 ± 0.23) mm vs. (2.16 ± 0.15) mm, P>0.05; 6 months after PCI, the LLL in DCB group was significantly smaller than that in DES group: (0.31 ± 0.28) mm vs. (0.48 ± 0.19) mm, and there was statistical difference (P<0.05). There were no significant differences in the incidences of myocardial infarction, TLR and cardiac death in DCB group (P>0.05). Before PCI and 6 months after PCI, there was no statistical difference in TpP between 2 groups (P>0.05); 1 month after PCI, the TpP in DCB group was significantly lower than that in DES group: (15.31 ± 6.13) mg/L vs. (19.46 ± 8.24) mg/L, and there was statistical difference (P<0.05). Conclusions DCB is an accurate and effective treatment for ACS patients with situ coronary artery disease under the intravascular ultrasound guidance, and it can reduce the risk of thrombosis.

Primary female urethral adenocarcinoma is rare. This paper reports a case of primary urethral mucinous adenocarcinoma complicated with signet ring cell carcinoma. The patient underwent urethral tumor resection in another hospital. Postoperative examination indicated that the tumor remained, and the tumor was completely removed after urethral tumor resection. After 11 months of follow-up, there was no tumor residue or recurrence.

OBJECTIVETo evaluate the safety and efficacy of fast track surgery (FTS) combined with laparoscopy in the treatment of infant Hirschsprung disease.

METHODSClinical data of 72 infants with Hirschsprung disease undergoing elective pull-through surgery from June 2010 to June 2013 were retrospectively summarized. The patients were divided into two groups: fast track surgery combined with laparoscopy group (FTS group, n=33) and laparoscopic surgery with traditional management perioperatively (control group, n=39). Postoperative intestinal function recovery, hospital stay, cost of hospitalization, complications were compared and postoperative recovery was followed-up for four weeks.

RESULTSThere were no significant differences in intraoperative blood loss and operative time between FTS and control group (both P>0.05). The recovery of bowel movement was earlier in the FTS group but the difference was not statistically significant (P=0.078). The hospital stay was shorter [(10 ± 2) d vs. (14 ± 4) d] and cost of hospitalization was lower [(15 316 ± 2273) Yuan vs. (18 641 ± 3082) Yuan] in FTS group than those in control group(P<0.01). Postoperative complications and recovery conditions during 4 weeks follow-up were similar between the two groups.

CONCLUSIONFast track surgery combined with laparoscopy in the treatment of infant Hirschsprung disease is safe and effective.

Objective: To evaluate the effect of intracoronary injection of recombinant human urokinase on plasma P-selectin in AMI patients with no-reflow during acute PCI. Methods: Ninety-two patients with acute ST-segment elevation myocardial infarction or acute non-ST-segment elevation myocardial infarction admitted to Center Hospital of Changchun City in January 2017 and December 2018 were randomly divided into two groups: 47 patients with intracoronary injection of sodium nitroprusside as control group and 45 patients with intracoronary injection of recombinant human urokinase as treatment group. Among them, 58 were males and 36 were females. The onset time was less than 12 h. The basic data, serum P-selectin, myocardial perfusion index and major adverse cardiovascular events were compared between the two groups. Results: In the treatment group, the corrected TIMI frame number, instant TIMI grade 3 blood flow, myocardial chromogenic grade 3 blood flow, myocardial necrosis marker CTnI, serum P-selectin were significantly lower than those in the control group: 31.26 ± 4.58 vs. 35.15 ± 6.25, 71.1%(32/45) vs. 51.1%(24/47), 64.4%(29/45) vs. 55.3%(26/47), (28.46 ± 3.95) ng/ml vs. (30.18 ± 3.47) ng/ml, (13.26 ± 4.58) ng/ml vs. (15.04 ± 3.98) ng/ml, and EF function was better. In the control group. The incidence of major adverse cardiac events in the treatment group was lower than that in the control group within one month after operation, but there was no statistical significance. Conclusions There is no reflux in patients with AMI during PCI. Intracoronary injection of recombinant human urokinase can improve myocardial perfusion without reflux and has no effect on fibrinolytic system in vivo. It does not increase the risk of systemic hemorrhage and the incidence of serious adverse cardiovascular events.

AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.