BACKGROUND:Recent studies have demonstrated that surgical trauma leads to lipid peroxidation in erythrocytes.However,injured erythrocytes play an important role in thrombosis following replacement.OBJECTIVE:To evaluate the influence of artificial total knee replacement on the lipid peroxdation in erythrocytes,and the prophylactic treatment of vitamin E and fructose 1,6 diphosphate(FDP)on it.DESIGN,TIME AND SETTING:Contrast clinical study,which was carried out in the Department of Otthopaedics,Affiliated Hospital of Nantong University from January 2003 to June 2006.PARTICIPANTS:Sixty patients with knee osteoarthritis who underwent artificial total knee replacement by anesthesia of epidural block were divided into four groups,including control group,vitamin E group,FDP group and vitamin E+FDP group,with 15 cases in each group.METHODS:Vitamin E was orally taken in the vitamin E group three days before replacement,three times a day,100 mg for each.The administration was performed until the surgical morning.Thirty minutes after the operation,FDP(10 g)was intravenously dripped in the FDP group.Additionally.vitamin E was also orally taken in the vitamin E+FDP group three days before replacement,three times a day,100 mg for each;on the surgical morning,FDP(10 g)was intravenously dripped on the first 30minutes.Blood samples were taken for biochemical determination before and after the operation at 1,3,5,and 7 days.MAIN OUTCOME M[EASURES:Corltents of malonaldehyde(MDA)and cuprum/zincum/superoxide dismutase (Cu-Zn-SOD)in the erythrocytes.RESULTS:MDA level in the vitamin E group and FDP group was significantly higher than that in the vitamin E+FDP group before and 5 and 7 days after replacement(P<0.05);while,Cu-Zn-SOD level was significantly lower(P<0.05).Otherwise,there were no significant differences in vitamin E+FDP group before and after replacement(P>0.05).CONCLUSION:The artificial total knee replacement can enhance lipid peroxidation and decrease antioxygen capability.However,the combination of vitamin E and FDP can prevent and relieve lipid peroxidation and antioxygen capability after replacement.

Objective To study the profiling and authentication of human cell lines in cell bank of our department using short tandem repeat (STR) loci and to analyze the situation of cell contamination and misjudgment. Methods Sixty-one human cells including cells collected and preserved by our cell bank and cells from the other departments were detected by the 16 STR loci-method. To analyze the cross contamina-tion between human cell lines, the results were aligned with profiles published by international cell banks. Isoenzyme detection was employed to authenticate the cell species when the STR signal can not be detected. Results Among the 61 cells, specific profiles were produced by 41 ceils and there was no cross contamina-tion. Thirty-six cells had the completely same STR profiles in 9 STR loci with the same cells preserved by ATCC or JCRB, while 5 cells have different profiling just in the vWA loci. The cells referred above can be recognized as correct cells; Eleven cells (18.0%) were the false cells. Among them, cancer cells of tongue named Tca8113 and cancer cell of liver named HHCC(changed to FHCC98 now) had the same profile with HeLa and HeLa S3 respectively; Two ceils both named HUT-102 have the completely different profiles with ATCC; The signal of 4 cells was not be detected, and all of them were determined as hamster cell lines by u-sing isoenzyme detection. Also 2 cells were identified to be mixed cells. Conclusion The phenomenon of cell misjudgment and cross contamination between cells is serious. Authentication of cell lines correctly, es-pecially for the re-authenticatian of domestic self-established cells, is very important for the guarantee of the reliability and reproducibility for scientific researches.

Objective To study the prevalence and genotype of human parvovirus B19 virus among blood products and plasma pools in China. Methods B19 DNA derived from 16 lots of factor Ⅷ concentrate produced by 4 manufactures and 10 lots of plasma pools were detected by nested PCR. Phylogenetic comparison of the partial B19 sequences obtained from positive samples were performed by direct sequencing. Results Twelve of sixteen lots of factor Ⅷ concentrate and all of ten lots of plasma pools were contaminated by B19 DNA. By comparing the partial B19 sequences,all the isolated viruses were genotype Ⅰ and their nucleotides were high conserved with homology of 98. 3%-100%. Conclusion B19 genotype Ⅰ DNA has been detected in high prevalence in factor Ⅷ concentrate and plasma pools. The genetic diversities were shown to be very low.

Objective To study the application of short tandem repeat (STR) profiling in quality control of human cell lines used for biological production. Methods The methods detecting 9 and 16 STR loci to identify human cell lines by PCR-capiilary electrophoresis were established respectively. Human cell lines, which were derived from many corporations and including diploid cell strains used for virus-vaccine production and 293 cell lines used for gene therapy products, were analyzed and compared by these two methods. Results The STR profiling methods used for authentication of human cell lines were established. Most of human diploid cell strains(20/21 ) used for virus-vaccine production from 13 corporations were iden-tiffed as the intended cells and no cross-contamination was found. However, one MRC-5 cells was identified as a false cell line and one MRC-5 had 3 alleles in D13S317 locus. For 12 strains of 293 cell lines, there were significant differences in STR profiling from different manufactures, which was likely be explained that the sources and gene modifications of these 293 cell lines are not well known and their genes are unstable during passage. Conclusion The STR profiling method has the advantages of high sensitivity and specifici-ty, and can be used for authentication of each of human cell lines for biological production.

Objective To optimize the extraction process of anti-hepatic fibrosis water-soluble component from Carthamus tinctorius L. via the orthogonal method. Methods The influences of type and content of solvent,extraction time,and extraction frequency on processing were investigated by assessing the content of hydroxysafflor yellow A,yield of extraction and anti-hepatic fibrosis potency via the high performance liquid chromatography and enzyme-linked immunosorbent assay by L9(3)4 orthogonal test. Results The optimum extracting condition for the anti-hepatic fibrosis water-soluble component from carthamus tinctorius L. was as follows: extracting with 10 times the volume of 10% ethanol for twice, 0. 5 h in each. Conclusion The process is scientific and feasible, which serves as a guideline for the production of the anti-hepatic fibrosis water-soluble components.

Objective To observe the effect of external use of Curcuma oil on the hyperplasia of mammary glands in rats and explore the mechanism. Methods Sixty female Wistar rats were randomly divided into following groups:normal con-trol group, model control group, Sanjierubi plaster group, low-, medium- and high-dose curcuma oil emulsion groups ( n=10 each).The models of hyperplasia of mammary glands were established by intramuscular injection of estradiol benzoate and proges-terone into the medial part of the rat hind limb.Different doses of medicines were given for 4 consecutive weeks.Enzyme-linked im-munosorbent assay, HE staining and immunohistochemistry were used to investigate the action mechanism of curcuma oil emulsion against mammary gland hyperplasia. Results Curcuma oil emulsion had preventive and therapeutic effects on the hyperplasia of mammary glands.The diameter of breasts was significantly reduced, the body weight restored, serum estradiol, follicle-stimulating hormone and prolactin levels profoundly decreased, progesterone, testosterone and luteinizing hormone levels markedly increased and the number and diameter of lobular acini obviously reduced in high-dose curcuma oil emulsion group when compared with those in model control group (P<0.05 or P<0.01). Conclusion Curcuma oil emulsion can remarkably improve the disturb-ance of serum hormones and inhibit the occurrence of hyperplasia of mammary glands.

Objective: To observe the clinical efficacy of self-made one-fourth tubular hook plate used in the fixation of comminuted fractures of acetabular posterior wall. Methods: The clinical data of 21 patients with comminuted fracture of acetabular posterior wall were retrospectively analyzed who had been treated by fixation with our self-made one-fourth tubular hook plate from June 2015 to September 2017 at Deparment I of Orhthopaedics, The Second People's Hospital of Anhui Province. They were 12 males and 9 females, aged from 20 to 59 years (mean, 38.5 years). The time from injury to surgery ranged from 3 to 6 days (average, 4.5 days). All the fractures were fixated with our self-made one-fourth tubular hook plate via the Kocher-langenbeck approach. Depending on the fracture fragments, the hook plate was cut out obliquely through the 3rd or 4th hole and the sharp end of the plate was bent into a small hook to hold the fragments. Only the hook plate was used in 6 cases and an additional construction plate in the other 15 cases. The postoperative reduction, functional recovery of the affected hip and complications were recorded. Results: The follow-up averaged 13 months (from 8 to 32 months). Intermuscular venous thrombosis occurred in one patient and sciatic nerve contusion was observed intraoperatively in one patient with sciatic nerve injury. According to the improved Matta criteria, the fracture reduction was rated as excellent in 16 cases, as good in 4 and as poor in one, giving a good to excellent rate of 95.2%. The fractures united after an average of 7.5 months (from 6 to 12 months). According to the improved Merle d'Aubigné and Postel scoring system, the hip function at the final follow-up was excellent in 14 cases, good in 3, fair in 2 and poor in 2, giving a good to excellent rate of 81.0%. Follow-ups revealed traumatic arthritis in 3 cases, femoral head necrosis in 2 cases and ectopic ossification in one. Conclusion Our self-made one-fourth tubular hook plate is simple, economical and effective because it can provide easy and rigid fixation of the comminuted fractures of acetabular posterior wall, leading to fine therapeutic effects.

The purpose was to optimize the vitrification for porcine embryos cryopreservation. Blastocyst/Morula (5-6th day-embryos) were collected from superovulated Bama mini-pigs (sows/gilts). We compared different cryopreservation methods, cryopreservation tools, thining of zona pellucida (ZP) and recipient breeds on the efficiency of porcine embryo cryopreservation. The results showed that: in embryo survival rate and blastocyst cell number, there were no significant differences between cryopreservation method I [embryos were vitrified by two step method with open pulled straw (OPS) and glass micropipette (GMP) in solution 1 (TCM199 + 20% FBS + 10% EG + 10% DMSO) for 3 min, and solution 2 (TCM199 + 20% FBS + 20% EG + 20% DMSO + 0.4 mol/L SUC) for 1 min, stored in liquid nitrogen] and method II[Blastocysts were cultured for 25 min in NCSU23 + 7.5 microg/mL cytochalasin B, centrifuged at approximately 13 000 xg for 12-13 min, and recovered back into pNCSU23. They were then equilibrated for 5 min in 2 mol/L ethylene glycol in pNCSU23, washed quickly in the vitrification medium, 8 mol/L ethylene glycol, 7% polyvinylpyrrolidone (PVP) in pNCSU23, loaded into OPS/GMP, and plunged into liquid nitrogen]. GMP vitrification method was more suitable and efficient than OPS method (P < 0.05) in embryo survival rate (83.8% vs 77.6%) and blastocyst cell number (53.1 vs 47.5) after thawing. Thining of ZP did not increase the survival rate, but significantly improved blastocyst cell number in the survival blastcysts (60.1 and 46, P < 0.01). Local pig breeds (Fengjing sows) were more suitable as recipients for embryo transfer of vitrified/warmed blastcysts, which can improve pregnant rate and embryo efficiency.
Animals ; Blastomeres ; cytology ; Cryopreservation ; methods ; veterinary ; Embryo Transfer ; veterinary ; Embryo, Mammalian ; Swine ; Swine, Miniature ; Vitrification ; Zona Pellucida ; physiology

Objective:To investigate the human milk oligosaccharides (HMOs) levels in breast milk of mothers delivering preterm infants and their effects on the early growth and development of infants.Methods:In this prospective cohort study, full-term and preterm newborns whose parents decided to breastfeed were recruited from Peking University Third Hospital between December 1, 2017 and November 30, 2018. The preterm infants were divided based on their gestational ages into extremely preterm (<28 weeks), very preterm (28-31 +6 weeks) and moderate to late preterm (32-36 +6 weeks) groups. Breast milk was collected from mothers at 7, 14, 28 and 120d postpartum. 368 breast milk samples were collected from 125 mothers in this study, including 54 mothers of full-term infants, 23 mothers of moderate to late preterm infants, 39 mothers of very preterm infants, and 9 mothers of extremely preterm infants. Ultra-performance liquid chromatography-mass spectrometer (UPLC-MS/MS) was used to determine the concentration of 2′-fucosyllactose (2′FL), 3-fucosyllactose (3FL), 3′-sialyllactose (3′SL), A-tetrasaccharide (P1), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-fucopentaose Ⅱ (LNFP-Ⅱ) and lacto-N-fucopentaose Ⅴ (LNFP-Ⅴ). Secretor status of mothers was defined as 2′-fucosyllactose (2′FL) concentration in colostrum and transitional milk greater than 200 μg/mL. Weight gain and the occurrence of allergic diseases of infants were collected at 120 d(4 months) postpartum. The chi-square test or the Fisher′s exact test was used for the comparison of categorical data between groups; Kruskal-Wallis test and Wilcoxon rank sum test were used for comparison of continuous data between groups. Nemenyi test was used for multiple comparison. Results:79.2% (99/125) of the mothers were secretor. There were no statistical differences between groups in the secretor status of mothers (χ2=1.31, P>0.05). The total concentration of HMOs peaked at 1-2 weeks postpartum. Compared to the preterm milk, the HMOs from the term milk was trending downwards at an earlier time. In the breast milk of secretor mothers on 28 d, total concentration of HMOs significant differed among the three groups of preterm milk and the term milk, with the median value of 4 587.09,4 615.25,5 277.44,5 476.03 μg/mL, respectively (Kruskal-Wallis χ2=8.1234, P=0.044). When analyzed by the median weight gain of the infants (low vs high weight gain) at 4 months postpartum, 2′FL was significantly lower in the high weight gain group at 7 d (1 818.04 μg/mL vs 2 181.67 μg/mL, W=1 386, P=0.018), while LNT & LNnT were significantly higher (1 182.36 μg/mL vs 1 053.62 μg/mL, W=816, P=0.044). The level of 3FL at 120 d was significantly affected by presence of allergic disease in infants, breast milk from mothers of infants with allergic disease had lower 3FL than those from mothers of infants without allergic disease (256.17 μg/mL vs 286.18 μg/mL, W=564, P=0.026). Conclusions:The overall profiles of HMOs in breast milk of mothers delivering preterm infants was basically the same as that of mothers delivering term infants; individual HMOs play a role in weight gain and the development of allergic diseases in preterm infants, but the mechanism is unclear and needs further study.

Objective:To investigate the human milk oligosaccharides (HMOs) levels in breast milk of mothers delivering preterm infants and their effects on the early growth and development of infants.Methods:In this prospective cohort study, full-term and preterm newborns whose parents decided to breastfeed were recruited from Peking University Third Hospital between December 1, 2017 and November 30, 2018. The preterm infants were divided based on their gestational ages into extremely preterm (<28 weeks), very preterm (28-31 +6 weeks) and moderate to late preterm (32-36 +6 weeks) groups. Breast milk was collected from mothers at 7, 14, 28 and 120d postpartum. 368 breast milk samples were collected from 125 mothers in this study, including 54 mothers of full-term infants, 23 mothers of moderate to late preterm infants, 39 mothers of very preterm infants, and 9 mothers of extremely preterm infants. Ultra-performance liquid chromatography-mass spectrometer (UPLC-MS/MS) was used to determine the concentration of 2′-fucosyllactose (2′FL), 3-fucosyllactose (3FL), 3′-sialyllactose (3′SL), A-tetrasaccharide (P1), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), lacto-N-fucopentaose Ⅱ (LNFP-Ⅱ) and lacto-N-fucopentaose Ⅴ (LNFP-Ⅴ). Secretor status of mothers was defined as 2′-fucosyllactose (2′FL) concentration in colostrum and transitional milk greater than 200 μg/mL. Weight gain and the occurrence of allergic diseases of infants were collected at 120 d(4 months) postpartum. The chi-square test or the Fisher′s exact test was used for the comparison of categorical data between groups; Kruskal-Wallis test and Wilcoxon rank sum test were used for comparison of continuous data between groups. Nemenyi test was used for multiple comparison. Results:79.2% (99/125) of the mothers were secretor. There were no statistical differences between groups in the secretor status of mothers (χ2=1.31, P>0.05). The total concentration of HMOs peaked at 1-2 weeks postpartum. Compared to the preterm milk, the HMOs from the term milk was trending downwards at an earlier time. In the breast milk of secretor mothers on 28 d, total concentration of HMOs significant differed among the three groups of preterm milk and the term milk, with the median value of 4 587.09,4 615.25,5 277.44,5 476.03 μg/mL, respectively (Kruskal-Wallis χ2=8.1234, P=0.044). When analyzed by the median weight gain of the infants (low vs high weight gain) at 4 months postpartum, 2′FL was significantly lower in the high weight gain group at 7 d (1 818.04 μg/mL vs 2 181.67 μg/mL, W=1 386, P=0.018), while LNT & LNnT were significantly higher (1 182.36 μg/mL vs 1 053.62 μg/mL, W=816, P=0.044). The level of 3FL at 120 d was significantly affected by presence of allergic disease in infants, breast milk from mothers of infants with allergic disease had lower 3FL than those from mothers of infants without allergic disease (256.17 μg/mL vs 286.18 μg/mL, W=564, P=0.026). Conclusions:The overall profiles of HMOs in breast milk of mothers delivering preterm infants was basically the same as that of mothers delivering term infants; individual HMOs play a role in weight gain and the development of allergic diseases in preterm infants, but the mechanism is unclear and needs further study.