Objective To study the repairment of pancreas mesenchymal stem cells(MSCs) on injured pancreas of rats,and find a new source of cells for treatment of diabetes. Methods ① Pancreases were taken out of three-day-old Wistar rats under bioclean condition,and cells were obtained by V-collagenase digestion. Generations were passed by conventional culture,cells were purified by adherence screening method,cell morphous was observed by Giemsa staining. The expressions of surface marker CD34 and CD44 were determined by FCM,and compared with bone marrow MSCs(BM-MSCs),the differences in character and function were observed. ②The experimental rats were divided into two groups randomly. Pancreas ischemic necrosis model was made by deligation,then the purified pancreas MSCs were marked with DAPI and then were transplantated partially. After two weeks,the survival rate was measured and histopathological detection was performed. Results ① The cells had concordant morphous gradually with vigorous generation. There was no significant difference in morphology and surface antigen compared with BM-MSCs.② The survival rate in experimental group was 75% ,the necrotic tissue had basi-rebounded. Blue fluorescent was observed in repaired pancreas tissues. The survival rate in control group was 20%. The survival rate in experimental group was higher than that in control group (P

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are conveniently cultured and separated in vitro because theirimmunogenicity is low. Therefore, BMSCs are suitable for cell transplantation. Research has shown that BMSCs are potential to repair neurological defect. OBJECTIVE: To determine whether in vitro cultured BMSCs can be transplanted to repair peripheral nerve injury or not, and to investigate its mechanisms. DESIGN, TIME AND SETTING: Randomized controlled animal study This study was performed in Department of Toxicology, Public Health College of Jilin University from March 2006 to March 2007.MATERIALS: Fifty healthy female Wistar rats aging 2 months and six 1-week-old female Wistar rats were used for extraction of BMSCs. Rabbit-anti-nerve growth factor (NGF) monoclonal antibody was provided by Santa Cruz Company. METHODS:BMSCs were separated and cultured with adherent method. In the 3rd generation, BMSCs were preiabeled with bromodeoxyuridine (BrdU) 48 hours before transplantation. Fifty healthy Wistar rats were selected to prepare sciatic nerve crush injury models with clamping method.Subsequently, rats were randomly divided into transplantation group and control group, with 25 rats in each group. Rats in the transplantation group underwent transplantation of BrdU-labeied BMSCs at nerve injured sites; while, the same volume DMEM was injected into rats in the control group. MAIN OUTCOME MEASURES: Injured nerve in the transplantation group suffered from anti-BrdU staining 1, 2, 4, and 6 weeks after surgery. Distal injured nerve in both groups suffered from NGF immunohistochemical staining 1, 2, 4, and 6 weeks after surgery. Image analysis system was adopted to analyze integrated absorbance of positive expression. Gait analysis was performed every week after surgery to measure sciatic nerve function index, and it was also adopted to measure regenerated nerve conduction velocity 6 weeks after surgery. Subsequently, amount and inner diameter of medullated nerve fibers were calculated after luxol fast blue staining, while wet weight of experimental-lateral gastrocnemius muscle and cross section area of muscle fiber were measured at the same time. RESULTS: Fifty rats were included in the final analysis. BrdU-labeled positive cells could be found at injured nerve in the transplantation group 1, 2, and 4 weeks after surgery. Integrated absorbance of NGF protein expression in the transplantation group was significantly higher than that in the control group 1 and 2 weeks after surgery (P < 0.01), but there were no significant differences between the two groups 4 and 6 weeks after surgery (P > 0.05). Sciatic nerve function index in the transplantation group superiorly recovered to that in the control group 3-6 weeks after surgery. Furthermore, 6 weeks after surgery, nerve conduction velocity, amount and diameter of medullated nerve fibers, wet weight and cross section area of gastrocnemius muscle in the transplantation group were significantly higher than those in the control group (P < 0.05-0.01). CONCLUSION: BMSCs can be transplantated into injuried nerve tissue, and promote the recovery of nerve function in the micro-enviroment, improve NGF expression in an early phase may be one of its mechanisms.

BACKGROUND: Studies are very few regarding the specific reaction of bone marrow mesenchymal stem cells (BMSCs) to activated microglia. Moreover, it remains unclear how MSCs maintain dopaminergic neuronal survival under specific microenvironment.OBJECTIVE: To explore the effect of BMSCs stimulated by activated microglia on dopaminergic neuron survival.METHODS: BMSCs were isolated from Wistar rats by attachment method, and in vitro cultured; microglia was activated, and dopaminergic neurons were cultured by enzyme digestion method. The experiment included 5 groups: BMSCs, microglia, lipopolysaccharide (LPS)+microglia; BMSCs+LPS+microglia groups, in which the dopaminergic neurons were cultured with corresponding culture medium; the dopaminergic neurons alone group was cultured with 10% fetal bovine serum+ DMEM/F12. The effect of different microenvironment on dopaminergic neuron survival and gliocyte-derived neurotrophic factor released from BMSCs were detected by immunofluorescence technique.RESULTS AND CONCLUSION: The release of gliocyte-derived neurotrophic factor in groups involving BMSCs was greater than corresponding control group. Tyrosine hydroxylase immunofluorescence showed that neuronal survival of dopaminergic neurons alone group was 15%, microglia group was 10%, LPS+microglia was 5%, but BMSCs+LPS+microglia group was 28%, significantly greater than the other groups (P < 0.05). In addition, survival of in vitro cultured dopaminergic neurons was decreased with increasing culture duration, but the survival of dopaminergic neurons in group involving BMSCs was significantly greater than corresponding control group. This indicates that microglia activation stimulated BMSCs to upregulate gliocyte-derived neurotrophic factor to prevent dopaminergic neurons from toxic injury, and inhibit delayed death of dopaminergic neurons.

Objective: To explore the influence of plateau environment in the development and morphology of the main organs of the offsprings of migrated rats, and to observe the pathological changes of heart, brain, and lung tissues of the offsprings of migrated rats. Methods: The 8-week Wistar rats who lived in the plain area were migrated to the plateau area. After 1 week, a total of 56 female and male rats were fertilized with a ratio of 3:1, and all the pregnant rats were natural childbirth. The offsprings rat pups were divided into three groups: 1 month, 3 months, and 6 months. Ten offsprings (5 female and 5 male) were randomly selected in each group to collect the heart, brain, lung, liver, kidney and other major organs to measure the weights. In each group, 45 offsprings were randomly selected to conduct water maze test, open field test and test of captive reaction. The brain, heart, and lung tissues from 5 offsprings in each group were collected for tissue section and the pathological changes of above organ tissues were detected with HE staining. Results: The pregnant rats moved from the plain to the plateau had normal feeding behavior, without preterm birth or death. On average, the pregnant rats had 8 to 10 babies per litter, with a total of 345 offsprings. The weight gain of offspring was about 1. 0-1. 5 g per day. Some of the offsprings had low intake and difficulty in foraging, and 15 offsprings died 3-5 d after birth, with a mortality rate of 4. 3%. The weight and the ratio of liver weight to body weight of the 6-month-old offsprings were increased compared with the 1-month-old offsprings (F<0. 05). In Morris water maze test there were no drowning and death in 45 offsprings at different time points; the time to find the water platform of 7 offsprings in 3-month-old group was lower than the others (P<0. 05), but there were no significant differences between various groups at other time points (P>0. 05). In open field test, there were 6 offsprings in open fieled test in 1-month old group showed a longer stay time in the central region than other rats (P<0. 05); there were no significant differences between various groups at the other time points (P>0. 05). In spite of all the time points in the captive reaction experiment, all the animals behaved in a similar way. The HE staining results of myocardium tissue showed the myocardial inflammatory cell infiltration, venous congestion, nucleus disappearance, and visible cellular outline in the 1-3 month old offsprings and the myocardial vascular congestion and myocardial space enlargement in the 6-month-old offsprings. The HE staining results of brain tissue showed the glass body formation and nucleus disappearance in the 1-month-old offsprings and neuronal cell body deformation, blood vessel congestion and vacuolar degeneration in the 3-6-month-old offsprings. The HE staining results of lung tissue showed the thicker alveolar walls, lymphocyte and plasma cell infiltration, pulmonary capillary expansion and congestion in the 1-3-month-old offsprings and the thicker alveolar walls, lymphocyte and plasma cell infiltration, pulmonary capillary expansion and congestion, pulmonary interstitial edema and red blood cell liquefaction in the blood vessels in the 6-month-old offsprings. Conclusion: Tibetan plateau environment has an influence in the development and morphology of heart, brain and lung of the migrated rats, and the reason may be related to low pressure and hypoxia of plateau.

Objective To optimize preparation process of Zushima Gel Cream. Methods The comprehensive evaluation set sensory evaluation, initial adhesive force, viscous force, and peeling strength score as indexes. The mixing time, refining temperature, mixing speed, and powder adding sequence were investigation factors. Orthogonal experiment was used to optimize forming process. Results Conditions of optimized preparation process were as following: add Zushima powder in Viscomate NP-700 and glycerol; mixing time was 5 min; refining temperature was 40 ℃; mixing speed was 100 r/min. Conclusion The preparation process is good and optimized Zushima Gel Cream has a good adhesive force, good glossiness and excipients. The preparation process is good.

Objective:To explore the effect of different doses of dexmedetomidine (Dex) on levels of tight-junction protein claudin-1 and diamine oxidase (DAO) in patients undergoing laparoscopic radical resection of gynecological malignant tumors.Methods:A total of 60 patients with gynecological malignant tumors who were scheduled to undergo laparoscopic radical resection under general anesthesia from January 2019 to January 2020 in the Second Hospital of Shanxi Medical University were selected, including 43 cases of cervical cancer (stageⅠ-Ⅱ A), 9 cases of ovarian cancer (stageⅠ A-Ⅲ C), and 8 cases of endometrial carcinoma (stageⅠ). Accroding to the random number table method, the patients were divided into control group (group C), low-dose Dex group (group D 1) and high-dose Dex group (group D 2), with 20 cases in each group. Patients in group D 1 were given Dex 0.5 μg·kg -1·h -1 by constant rate intravenous infusion pump after induction until 30 min before the end of operation. Patients in group D 2 were given Dex 1.0 μg·kg -1·h -1 by constant rate intravenous infusion pump after induction until 30 min before the end of operation. Group C adopted the same calculation method and received the same amount of 0.9% sodium chloride solution by infusion pump. At 10 min before induction (T 1), 1 hour after pneumoperitoneum (T 2) and 12 hours after pneumoperitoneum (T 3), 5 ml of brachial vein blood was collected from the patients, and the levels of claudin-1 protein, DAO and blood glucose were measured. Results:At T 1, T 2 and T 3, the expression levels of claudin-1 in group C were (77.05±17.61) pg/ml, (66.76±12.97) pg/ml and (55.93±12.71) pg/ml, and the difference was statistically significant ( F = 10.449, P<0.05); the expression levels of DAO in group C were (4.83±0.93) ng/ml, (5.62±1.01) ng/ml and (5.98±1.21) ng/ml, and the difference was statistically significant ( F = 6.139, P < 0.05); the levels of blood glucose in group C were (4.82±0.66) mmol/L, (7.55±0.94) mmol/L and (6.51±0.54) mmol/L, and the difference was statistically significant ( F = 70.197, P < 0.05). At T 2, the expression level of claudin-1 in group D 1 was (69.12±13.02) pg/ml, which was not significantly different from group C ( t = -0.575, P > 0.05); the expression level of claudin-1 in group D 2 was (76.36±14.89) pg/ml, which was higher than that in group C, and the difference was statistically significant ( t = -2.175, P < 0.05). At T 3, the expression levels of claudin-1 in group D 1 and group D 2 were (66.14±14.36) pg/ml and (73.37±16.93) pg/ml, which were higher than that in group C, and the differences were statistically significant ( t values were -2.380 and -3.682, both P < 0.05). The expression levels of DAO in group D 1 and group D 2 were (5.02±0.84) ng/ml and (4.91±0.93) ng/ml at T 2, and (5.29±0.86) ng/ml and (5.20±0.98) ng/ml at T 3, which were lower than those in group C, and the differences were statistically significant ( t values were 2.051, 2.295, 2.079 and 2.285, all P < 0.05). The levels of blood glucose in group D 1 and group D 2 were (7.10±0.66) mmol/L and (6.77±0.97) mmol/L at T 2, and (5.95±0.94) mmol/L and (5.93±0.74) mmol/L at T 3, which were lower than those in group C, and the differences were statistically significant ( t values were 2.565, 5.374, 2.293 and 2.765, all P < 0.05). Conclusion:Continuous infusion of Dex can inhibit the stress response caused by long-term CO 2 pneumoperitoneum in laparoscopic radical resection of gynecological malignant tumors, and adjust the changes of expression levels of claudin-1 protein and DAO, reduce the damage of intestinal mucosal cells, facilitate the recovery of intestinal function, and the effect of high-dose Dex is better than low-dose Dex.

In order to improve the yield of bacterial cellulose (BC), the fermentation medium of BC-producing strain J2 (Gluconobacter) was optimized, and BC ultra-micro-structure was observed. Initially, Plackett-Burman design was employed to evaluate eight variables which were relevant to BC production. Three statistically significant parameters including yeast extract, ZnSO4, ethanol were selected and other 5 variables were not significant (P > 0.05). The optimized levels of three variables were defined by Box-Behnken design and response surface methodology (RSM). BC ultra-micro-structure was observed by scanning electron microscope (SEM) with cotton cellulose as comparison. The results indicated that the BC yield under the optimum fermentation medium was 11.52 g/100 mL, which was as 1.35 times as that under the original fermentation medium. The SEM photos manifested that bacterial cellulose ribbon, with a diameter less than 0.1 microm, was less than cotton cellulose ribbon. The bacteria inside the cellulose net were eliminated after the NaOH treatment.
Cell Culture Techniques ; Cellulose ; biosynthesis ; ultrastructure ; Culture Media ; chemistry ; Fermentation ; Gluconobacter ; cytology ; metabolism

Objective To investigate the antibiotic resistance in clinical isolates in the Second Hospital of Hebei Medical University from 2015 to 2016.Methods A total of 16 292 strains of non-duplicate bacterial strains were collected.The isolates were subjected to identification and antimicrobial susceptibility testing on VITEK 2-Compact system.The data were processed and analyzed using WHONET 5.6 software.Results Specifically,7 961 and 8 331 strains of pathogens were collected in 2015,2016,respectively.Gram-negative bacteria accounted for 62.0％ in 2015 and 66.9％ in 2016,respectively.The top five pathogens isolated in these two years were still Klebsiella pneumoniae,Escherichia coli,Acinetobacter baumannii,Pseudomonas aeruginosa,and Staphylococcus aureus.The proportion of K.pneumoniae increased to the first place in 2016 which accounted for 16.1％.Coagulase negative Staphylococcus in blood samples decreased from 42.6％ in 2015 to 30.0％ in 2016.Vancomycin resistant strains were not found in Staphylococcus.In 2015 and 2016,the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) was 56.2％ and 51.3％,respectively.The prevalence of methicillinresistant coagulase negative Staphylococcus (MRCNS)was 79.3％ and 63.1％,respectively.Enterococcus faecalis and Enterococcus faecium accounted for 25.2％ and 73.2％ respectively in the 911 strains of Enterococcus.In 2015 and 2016,3.1％ and 2.9％ of the E.faecium strains were resistant to vancomycin,respectively.In 2016,the prevalence of carbapenemresistant strains increased in K.pneumoniae,E.coli and E.cloacae.K.pneumoniae showed increasing resistance rate to all the antimicrobial agents tested except for gentamicin and amikacin.The percentage of the K.pneumoniae strains resistant to imipenem and meropenem increased from 19.3％,18.5％ in 2015 to 24.2％,23.1％,respectively.A.baumannii isolates were still highly resistant to the commonly used antibiotics in these two years,but relatively susceptible to polymyxin B,tigecycline,cefoperazone-sulbactam and minocycline (＜30％ resistant).P.aeruginosa isolates showed lower resistance rate to amikacin (11.7％),cefoperazonesulbactam (15.5％),piperacillin-tazobactam (18.7％),ceftazidime (20.1％),cefepime (21.9％).P.aeruginosa presented a trend of declining resistance to all the antimicrobial agents tested from 2015 to 2016,except aztreonam,to which the resistant P.aeruginosa strains increased from 27.0％ in 2015 to 34.7％ in 2016.Conclusions The antimicrobial susceptibility profile of clinical bacterial isolates has been changing constantly.We need to adopt effective infection prevention and control measures in hospital and further standardize and control the use of antibacterial agents.

OBJECTIVE To establish a quality standard for rice-fired Glehnia littoralis . METHODS Appearance observation ， powder microscopic identification and thin-layer chromatography （TLC）identification were performed for the samples of rice-fired G. littoralis decoction piece. According to the relevant methods stated in 2020 edition of Chinese Pharmacopoeia （part Ⅳ），the contents of moisture ，total ash ，acid-insoluble ash ，water-soluble extract and acid-soluble extract were determined. The contents of psoralen，zanthoxylin，bergapten，imperatorin and isoimperatorin were determined by high performance liquid chromatography （HPLC）. RESULTS The rice-fired G. littoralis decoction pieces were round-like small segments ，slightly rough ，yellow（peeled） or dark yellowish brown （with peel ），special gas and slightly sweet taste. The powder was yellowish white. Under the microscope ， secretions and secretory cells ，ducts，gelatinized starch granules ，ray cells ，parenchyma cells ，etc. could be seen. TLC showed the spots developed clearly. In the chromatogram of the test sample ，there was the same blue fluorescent spot at the corresponding position of the chromatogram of isoimperatorin control sample. The moisture ，total ash ，acid-insoluble ash ，water-soluble extract and ethanol-soluble extract from 9 batches of samples were 5.82％-6.27％，3.19％-3.59％，0.21％-0.27％，24.91％-30.30％ and 20.66％ -25.83％ ，respectively. The linear range of psoralen ，zanthotoxin，bergapten，imperatorin and isoimperatorin were 0.240-2.400，0.320-3.200，0.224-2.240，0.292-2.920，0.208-2.080 µg/mL（all r＞0.999 0）. Limits of quantitation were 0.032 0， 0.030 0，0325 0，0.032 0，0.045 0 µg，respectively. Limits of detection were 0.100 8，0.089 6，0.071 5，0.090 0，0.132 0 µg， respectively. RSDs of prescision ，stability（24 h）and reprodu- cibility tests were less than 3％. Average recoveries were 100.56％ （RSD＝1.36％ ，n＝6），100.73％（RSD＝2.25％ ，n＝6）， 100.36％（RSD＝0.98％，n＝6），98.24％（RSD＝0.40％，n＝6） E-mail：853063968@qq.com and 99.40％（RSD＝0.35％，n＝6），respectively. The contents of above five components were 5.85-13.31，8.63-33.38，6.23- E-mail：shixiaofeng2005@sina.com 15.25，6.12-12.98，5.52-10.77 µg/g，respectively. The total contents were 34.20-83.47 µg/g. CONCLUSIONS It is preliminarily proposed that the moisture ，total ash and acid-insoluble ash should not exceed 7.30％，4.10％，0.30％. The water-soluble extract and ethanol-soluble extract are no less than 21.00％ and 18.00％，respectively. The total content of coumarin should not be less than 52.03 µg/g（with peel ）and 26.34 μg/g（peeled）. Established quality standard can be used for the quality control of rice-fired G. littoralis .

Objective:To investigate the therapeutic effect of dorsal skin-tightening technique on the correction of mild penile curvature in children with hypospadias.Methods:The clinical characteristics of hypospadias patients (95 cases) with mild penile curvature (<30°) after degloving penis during operation in our hospital from Jan 2017 to Sep 2020 were analyzed retrospectively. Group A: A new technique, reconstructing penile pubic angle at 12 o'clock position of penile dorsal side after degloving and suturing forskin outer and inner plate with tension at 12 o'clock position, was performed in dorsal skin-tightening group (43 cases). Gtoup B: while in dorsal midline tunica albuginea plication group (52 cases), Buck fascia was incised on dorsal midline area, following by tunica albuginea plication with one or two stitches. The patients in Group A were 0.4 to 1.5 years old, and the median age was 1.1 years, urethral orifice were located on distal shaft in 36 cases, proximal in 7 cases.The patients in Group B were 0.5 to 2.6 years old, and the median age was 1.5 years, urethral orifice were located on distal shaft in 41 cases, proximal in 11 cases. The penile ventral curvature degree was recorded during regular follow-up (postoperative 6 and 12 months), as well as postoperative complications.Results:Artificial erection test showed penile curvature was corrected during surgery by measuring with protractor. There was no chordee by measuring with the side photos in all patients during an average of 1.6 years follow-ups. There were 4 case of urethral fistula in Group B and 3 cases in Group A. No cases of urethrostenosis, diverticulum or concealed penis was recorded. The difference of postoperative complications had no statistical significance.Conclusion:Hypospadias with mild penile curvature could be effectively corrected by dorsal skin-tightening technique, which showed a good result of early follow-up.