Objective To discriminate whether chemical compounds are micronuclei-inducing by counting the ratio of bone marrow micronucleus polychromatic erythrocytes ( MNPCE) dyed with a single fluorescence reagent ( acriding organe, AO) by single-laser flow cytometry.Methods Treating male KM mice and SD rat with cyclophosphamide ( CP) respectively, and counting their frequencies of micronucleated polychromatic erythrocytes ( MNPCE) as well as frequencies of micronucleated normochromatic erythrocytes ( MNNCE ) in bone marrow by AO and a single_laser flow cytometer (FCM), comparing and analyzing the results from different methods.Results The results showed that, along with increasing dose of CP, the ratio of MNPCE also corresponding increase, suggesting significant quantative-efficiency correlation.MNPCE were also counted manually by fluorescence microscopy, and the results showed no significant difference with that by flow cytometry.Conclusions AO_FCM fully automated detection method for the detection of MNPCE and MNNCE in mice and rat bone marrow micronucleus rate is reliable.

OBJECTIVE:To evaluate inpatients' use of antibiotics in our hospital.METHODS:The inpatients' use of antibiotics in our hospital during 2004～2006 was analyzed retrospectively in respect of the consumption sum and DDDs etc.RES-ULTS:The proportion of antibiotics in the total western medicines were decreasing year on year in terms of consumption sum.Cephalosporin,quinolones,lincomycins and penicillins had been dominated the first 4 places during the 3 consecutive years.Leading the list in terms of consumption sum and DDDs was Clindamycin injection during the 3 consecutive years.CONCLUSION:The inpatients' use of antibiotics was characterized by high starting point,excessively depending on empirical treatment,which needs to be further monitored to achieve a rational use of antibiotics.

Objective To establish a HPLC method for the determination of psoralen and bergapten in Ficus hirta Vahl.Methods A Merck-lichrospher C18(4 mm?250 mm,5 ?m)column was adopted.The mobile phase was acetonitrile-water(35∶65) with the flow rate of 1.0 mL/min.The column temperature was 35 ℃,and the detection wavelength was set at 222 nm.Results Psoralen's linearity was obtained in the range of 0.12 ?g～1.20 ?g(r=0.999 7),and bergapten's linearity in the range of 0.03 ?g～0.30 ?g(r=0.999 5).Psoralen's average recovery was 100.9 %with RSD 2.9 %,and that of bergapten was 99.6 %with RSD 1.9 %.Conclusion This is the first report of simultaneous determination of psoralen and bergapten in Ficus hirta Vahl.by HPLC,and the results showed the method is accurate,reproducible,and can serve as quality evaluation method for Ficus hirta Vahl.

Objective To investigate an accurate and applicable localizing method of puncture point for neuroendoscopic third ventriculostomy in neurosurgery so as to ensure that the neuroendoscope is led to the anterior membrane of mammillary body on the base of third ventricle directly. Methods Based on the MR characteristics including multiple directions and angles and high soft tissue resolution, the coordinate of puncture point was measured and calculated for neuroendoscopic third ventriculostomy in MRI. Results~The position of puncture point for neuroendoscopic third ventriculostomy was (127.2?9.9)mm to nasal root, or (17.1?5.6)mm in front of coronal suture and (20.3?4.7)mm to sagittal suture. The angle between the puncturing line and the cerebral falx was (12.3?1.9)?, and the depth from scalp to anterior membrane of mammillary body was (89.3?10.4)mm. Conclusion MRI-guided localization for neuroendoscopic third ventriculostomy is an accurate, simple, safe, painless, and applicable method.

ObjectiveTo investigate the efficacy of combined therapy of mild hypothermia and hibernation to treat severe brain injury. Methods24 patients with severe brain injury were randomly divided into combined therapy group and normothermia group. Glasgow Coma Scale scores of all the patients were in the range of 3 to 8. No later than 10 hours after their injury, hypothermia patients were given half dosage of No.1 hibernation cocktail and had been cooled by cooling blankets to 32℃-34℃ (rectal temperature) for 5 days, then to 35℃ for 24 hours, and slowly increased to their normal level. 3 days and 7 days after their admission, intracranial pressure,creatine phosphate kinase,partial pressure of arterial O2 and CO2, platelet and Na+,K+ were measured.7 days after their admission, Glasgow Outcome Scale scores of each patient and mortality of each group were measured. ResultsThe mortality of combined therapy group(25.0%) was significantly lower than that of normothermia group (66.6%,P<0.05). The decreased values of intracranial pressure, creatine phosphate kinase and platelet number of combined therapy group were all significantly higher than that of normothermia group respectively (P<0.05). There were no significant difference in mean artery pressure, blood electrolyte, and partial pressure of arterial O2 and CO2 between these two groups(P>0.05). ConclusionThe combined therapy of mild hypothermia and hibernation can effectively reduce the mortality of patients with severe brain injury as it is much easier, less invasive and with less complications.

Objective To investigate the effects and value of enhance recovery after surgery (ERAS) application in patients with hepatocellular carcinoma underwent liver resection.Methods One hundred and seventy-two patients with hepatocellular carcinoma underwent liver resection in the first affiliated hospital of Harbin medical university from June 2013 to June 2015,ERAS group (92 cases) and control group (80 cases) were retrospectively studied.Laboratory indicators (ALT,AST,TBIL,ALB,PA,lymphocyte count),postoperative complications,postoperative hospitalization days and total hospitalization expenses were compared between two groups.Results For postoperative liver functional indicators (ALT,AST,TBIL),ERAS group Day 1 and Day 7 were respectively (216.3±141.7) U/Land (70.1 ±29.4) U/L,(184.0±155.8) U/Land (39.1 ±17.5) U/L,(22.4± 8.7) μmol/L and (20.0 ± 7.5) μmol/L,control group were respectively (304.5 ± 226.2) U/L and (83.9 ± 48.5) U/L,(294.1 273.0) U/L and (49.2 ±33.8) U/L,(26.9 ±15.6) μmol/L and (24.6 ± 10.8) μmol/L,the difference between two groups was statistically significant (F =9.33,9.84,9.26,P ＜ 0.05).For postoperative nutritional indicators (ALB,PA),ERAS group Day 7 were respectively (35.3 ± 4.4) g/Land (136.3 ±34.1) mg/L,control group were respectively (33.6 ±4.2) g/L and (108.0 ± 32.5) mg/L,the difference was statistically significant (F =4.97,4.54,P ＜ 0.05).For postoperative immune indicators (lymphocyte count),ERAS group Day 1 and Day 7 were respectively (0.9 ±0.3) × 109/L and (1.5 ± O.5) × 109/L,control group were respectively (0.7 ± 0.3) × 109/L and (1.3 ± 0.5) × 109/L,the difference was statistically significant (F =7.37,P ＜ 0.05).For postoperative complications (hemorrhage,bile fistula,hepatic dysfunction,infection) were no statistically significant differences (P ＞ 0.05),however,ascites had statistically significant difference (x2 =7.609,P ＜ 0.05).Off bed time,postoperative exhaust time,postoperative hospitalization time and total hospitalization expense of ERAS group were respectively (1.7 ± 0.5) days,(2.3 ± 0.6) days,(9.8 ± 2.3) days,(4.6 ± O.9) × 104 RMB,control group were respectively (3.0 ± 0.7) days,(3.4 ± 0.8) days,(17.6 ± 5.8) days,(6.3 ± 2.1) × 104 RMB,the difference was statistically significant (t =13.032,10.937,11.371,7.118,P ＜ 0.05).Conclusions Application of ERAS in patients with HCC underwent liver resection is safe and effective.ERAS effectively reduce stress reaction of patients,promote the recovery of liver function,improve the postoperative immune and nutrition status,shorter postoperative hospitalization time,and reduce the total cost of hospitalization.

Objective To explore the effective measurements of integrated therapy for the ear keloids in the Oriental. Methods Surgical excision was performed for keloid, and the cut then sutured directly or with flap transfer. At early stage of surgery (within 24 hours), X-ray or electron-ray radiotherapy was used for 3-5 times once daily; and steroids injected after cutting lines at 7th day every two weeks and gradually ended. Results A total of 48 patients were treated, and followed, in which 12 patients were lost of follow-up. The total effective rate was 94. 4 % based on the data of follow-up in 36 patients for 1-3 years. Therefore, the curation rate was 83. 3 % (30 cases) and effective rate 11. 1 % (4 cases). Conclusions Surgical cutting combined with early radiotherapy and steroids injection is the effective integrated therapy for the ear keloids in the Oriental.

Objective To study the delayed gastric emptying for patients underwent pylorus-preserving pancreaticoduodenectomy and standard Whipple procedure.Methods Clinical data of 401 consecutive patients who underwent standard Whipple procedure/pylorus-preserving pancreaticoduodenectomy between January 2012 and July 2016 in the First Affiliated Hospital of Harbin Medical University were retrospectively collected and analyzed.Using the independent-samples t test,x2 test or Fisher's exact test,Mann-Whitney test and other statistical methods to compare the postoperative complications between pylorus-preserving pancreaticoduodenectomy and standard Whipple procedure group.Results Compare with standard Whipple procedure group,delayed gastric emptying after pylorus-preserving pancreaticoduodenectomy occurred in 8 of 35 patients (22.9％),obviously higher than it after standard Whipple procedure occurred in 40 of 366 patients (10.9％) (P =0.038).Other postoperative complications were not significantly different (P ≥ 0.05).Each level of delayed gastric emptying after pyloruspreserving pancreaticoduodenectomy and standard Whipple procedure were not significantly different (P ≥ 0.05),but the average recovery time of delayed gastric emptying after pylorus-preserving pancreaticoduodenectomy (12.13 ± 3.09) d was obviously shorter than it after standard Whipple procedure (17.28 ± 9.63) d (P =0.009).Conclusions Pylorus-preserving pancreaticoduodenectomy increases the risk of delayed gastric emptying,but it does not increase severity delayed gastric emnptying by each level.The recovery time of delayed gastric after pyloruspreserving pancreaticoduodenectomy is shorter.

BACKGROUND:Rheumatoid arthritis is an autoimmune disease, and traditional treatment methods are difficult to effectively solve the patient's lack of immune tolerance mechanisms. With the development of stem cel s in regenerative medicine, stem cel therapy has become a hot spot in the treatment of autoimmune diseases.
Currently, studies on cel transplantation for the treatment of rheumatoid arthritis are rarely reported.
OBJECTIVE:To study the influence of umbilical cord mesenchymal stem cel therapy on the changes of Th1/Th2 and Treg in rheumatoid arthritis patients, thereby seeking new therapies for rheumatoid arthritis.
METHODS:We selected 180 cases of rheumatoid arthritis, including 27 patients as control group undergoing non-steroidal anti-inflammatory drugs and anti-rheumatic drugs and 153 patients as cel treatment group undergoing intravenous infusion of 40 mL umbilical cord mesenchymal stem cel s at a density of 4×107. Dosing regimen was same in the two groups. The 76 of 153 patients accepted second cel therapy at 3-4 months after the first cel therapy. After fol ow-up of 3 and 6 months, clinical effectiveness evaluation (DAS28, HAQ, ACR20), rheumatoid factor, anti-CCP antibodies, T cel subsets, Th cytokine were detected;for patients with second cel therapy, T cel subsets and Treg were detected at 8 months after treatment.
RESULTS AND CONCLUSION:(1) At 3 months after treatment, the DAS28, HAQ and ACR20 scores were significantly lower in the cel treatment group than the control group (P<0.01). (2) At 3 and 6 months after cel therapy, the DAS28 and HAQ scores were significantly decreased in the cel treatment group (P<0.01), and these scores were decreased continuously after second cel therapy (P<0.01). (3) Interferon-γlevel in cel s did not change obviously at 3-6 months after treatment, but the interleukin-4 level was gradual y increased at 6 months after treatment (P<0.05). (4) The number of Treg cel s was significantly increased at 3-6 months after treatment (P<0.01), which was closely related to ACR, especial y ACR70 percentage (P<0.05);the ratio of CD4+Treg was increased significantly at 3 months after treatment (P<0.05), and this increasing trend was also maintained at 6 and 8 months after treatment, but there was no significant difference (P>0.05). (5) B cel levels were significantly decreased at 6 months after treatment (P>0.05);the rheumatoid factor value was significantly decreased at 3-6 months after treatment (P<0.05). (6) There was no change in anti-CCP antibody and interleukin-17 levels at 3-6 months after treatment. These findings indicate that after treatment with umbilical cord mesenchymal stem cel s, the Th1/Th2 tends to balance and Treg level is elevated in rheumatoid arthritis patients, which are directly related to clinical trials and symptomatic relief. Therefore, standard rheumatism medication combined with umbilical cord mesenchymal stem cel transplantation can improve immune network effects, adjust the immune tolerance and prevent il ness progress in rheumatoid arthritis patients.

To study the umbilical cord mesenchymal stem cells impact of inflammatory factors in rheumatoid arthritis patients.Methods:94 cases of patients with RA hospitalized in our department from April 2011 to December 2012 were treated with umbilical cord mesenchymal stem cells (MSCs) UC,during which the cell therapy scholastic ethics was committed approvally and patients′informed consents were separately signed .94 patients were directly intravenous infusion with 40 ml UC-MSCs ( 4×107 cells/ml),including 57 cases with two UC-MSCs therapy.We used multifunctional flow lattice Luminex 200 analysis to detect contents of 13 kinds of factors in serum such as TNF alpha,IFN-gamma,IL-1β,IL-4,IL-6,IL-8,IL-10,ie,and detected CPR,ESR,assessment DAS28,HAQ,and ARA.Follow-up treatments were performed after 1 week,3 months,6 months ( two cells treatment of 60 cases).Results:①DAS28,HAQ grading standards,were decreased (P<0.01) in 3 months and 6 months before the patients treatment ,2 times than one continue treatment decreased ( P<0.01 );the ESR and CRP in 1 week dropped significantly after treatment ( P<0.01),3 months,6 months before the treatment also decreased (P<0.05).②IL-6,TNF alpha were in falling levels after 1 week,3 months and 6 months treatment ,( P<0.05 ).Conclusion: We proved the inflammatory factors directly related with clinical indicators and symptoms of RA patients .94 cases of patients with other inflammatory factor (IL-17,IL-4,IL-10,etc.) also had some change,we still needed further observation.According to drug rheumatism guide at the same time , collaborative using with UC-MSCs could make RA patients improve local and systemic inflammatory response ,prevent disease progression.