Objective For bioequivalence test of the consistency evaluation of generic drug products,providing a reference of varieties of biowaiver.Methods Based on Human bioequivalence test waiver guidelines (draft),on condition that first drug of the consistency evaluation,to introduce and conclude briefly the standards of biowaiver and varieties of biowaiver in FDA,WHO and EMA.Results Contrast to FDA,there are 59 varieties applied for the waiver and 19 varieties not applied for the waiver in the 289 varietie;compared to WHO,10 drugs are exempted and 1 grug is exempted in EMA.Conclusion At present,the specific list of drugs are not published of biowaiver in our country,the pharmaceutical companies should compare and consult revelant standards and specific drugs in China and abroad,to speed up the progress of the consistency evaluation.

Sugarcane molasses containing large amounts of sucrose is an economical substrate for succinic acid production. However, Escherichia coli AFP111 cannot metabolize sucrose although it is a promising candidate for succinic acid production. To achieve sucrose utilizing ability, we cloned and expressed cscBKA genes encoding sucrose permease, fructokinase and invertase of non-PTS sucrose-utilization system from E. coli W in E. coli AFP111 to generate a recombinant strain AFP111/pMD19T-cscBKA. After 72 h of anaerobic fermentation of the recombinant in serum bottles, 20 g/L sucrose was consumed and 12 g/L succinic acid was produced. During dual-phase fermentation comprised of initial aerobic growth phase followed by anaerobic fermentation phase, the concentration of succinic acid from sucrose and sugarcane molasses was 34 g/L and 30 g/L, respectively, at 30 h of anaerobic phase in a 3 L fermentor. The results show that the introduction of non-PTS sucrose-utilization system has sucrose-metabolizing capability for cell growth and succinic acid production, and can use cheap sugarcane molasses to produce succinic acid.
Bioreactors ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fermentation ; Membrane Transport Proteins ; genetics ; Metabolic Engineering ; Molasses ; Saccharum ; chemistry ; Succinic Acid ; chemistry ; Sucrose ; chemistry

PURPOSE: To produce animal models of Acanthamoeba keratitis and to evaluate the advantages and adaptation range of each of the three methods employed. MATERIALS AND METHODS: Mice and Wistar rats in three groups of 15 rats and 15 mice each were used to establish the models. Right corneas in group A were scratched and challenged with Acanthamoeba. Those in group B were scratched and covered with contact lenses incubated with Acanthamoeba. Those in group C received an intrastromal injection of Acanthamoeba. Five rats and 5 mice in each group were used for histopathological investigations and the other 10 in each group were used for clinical evaluation. The models were evaluated by slit lamp examination, microscopic examination and culture of corneal scrapings, HE staining of corneal sections, and pathological scoring of the infections. RESULTS: Four rats and 6 mice in group A, 7 rats and 8 mice in group B, and 10 rats and 10 mice in group C developed typical Acanthamoeba keratitis. CONCLUSION: Corneal scratching alone has the lowest infection rate, while scratching and then covering with contaminated contact lenses has a moderate rate of infection and most closely mimics what happens in most human infections. Intrastromal injection of Acanthamoeba gives a much higher infection rate and more severe Acanthamoeba keratitis.
Acanthamoeba/growth & development ; Acanthamoeba Keratitis/*parasitology/pathology ; Animals ; Contact Lenses/adverse effects/parasitology ; Cornea/parasitology/pathology ; Disease Models, Animal ; Female ; Male ; Mice ; Microscopy ; Rats ; Rats, Wistar

Objective To investigate the phenotypic and genetic characteristics of the lysostaphin‐resistant Staphylococcus aureus variants induced by recombinant lysostaphin in vitro .Methods Three clinical isolates of S . aureus ,including two resistant to methicillin (MRSA ) and one susceptible to methicillin (MSSA ) were induced by treatment with sub‐MIC of recombinant lysostaphin via one‐step selection in vitro .Susceptibility of the variants to antibiotics were determined and compared with their parental strains .The full length of femABX genes was amplified by polymerase chain reaction and sequenced to identify the potential mutation sites in these genes .The growth‐curve in liquid medium and virulence in a mouse systemic infection model of both parental and variant strains were observed . Results The frequency of lysostaphin resistance in S . aureus was between 10-4 to 10-8 following induction by lysostaphin . Resistance to lysostaphin was associated with a significant decrease in growth rate in vitro and virulence in vivo ,as well as increased susceptibility toβ‐lactams evidenced by the M IC of β‐lactams against the variants as low as 1/4 000 to 1/2 of the M IC against their parental strains . Sequencing of f emA BX genes showed mutation in femA gene in both variants ,which resulted in a premature termination codon .Conclusions Resistance of S . aureus to lysostaphin may develop following induction by recombinant lysostaphin in vitro . The lysostaphin‐resistant S . aureus variants are characteristic of lower growth rate , decreased virulence ,and higher susceptibility to β‐lactams .

Objective: To investigate bone mineral density of college students with masked obesity, and to provide theoretical basis for bone density improvement and osteoporosis prevention in college students. Methods: Participants were enrolled from universities and colleges. A total of 518 low-weight or normal-weight students were included and were classified according to the criteria of masked obesity, dual-energy X-ray test was used to detect the percentage of body fat and bone mineral density of college students in each group. The differences in bone mineral density between masked obesity and non-masked obesity groups of different genders were compared, and the correlation between body mass index, body fat percentage and bone mineral density was analyzed. Results: Among the low-weight male college students, the whole body bone mineral density, t-value and Z-value of masked obesity group were lower than those of the nonmasked obesity group [(1.82±0.04,2.01±0.22)g/cm 2; (-1.10±0.24,-0.02±0.15); (-0.94±0.64,-0.01±0.43)(P<0.01)]. Among low-weight male college students, bone mineral density of the upper limbs, thighs, ribs, and pelvis of the masked obesity group was lower than that of the non-masked obesity group. Among low-weight female college students, body bone mineral density of masked obesity group was lower than that of the non-masked obesity group [(1.13±0.48,1.31±0.29)g/cm 2; (P<0.05)]. Among low-weight female college students, bone mineral density of head, upper limbs, thighs, and trunk of masked obesity group was lower than that of the nonmasked obesity group. In male college students, body mass index was positively correlated with bone mineral density, T-value and Z-value (r=0.69, 0.68, 0.61, P＜0.01) while body fat percentage was negatively correlated with bone mineral density, T-value and Z-value (r=-0.52, -0.51, -0.49, P＜0.01). In female college students, body mass index was positively correlated with T-value and Z-value (r=0.46, 0.26, P＜0.01), and body fat percentage was negatively correlated with T-value and Z-value (r=-0.22, -0.23, P＜0.01). Conclusion Compared with normal-weight college students, depletion of bone mineral density among masked obesity students is observed. It is of great significance to pay attention to bone mineral density of college students with masked obesity and implement intervention to prevent osteoporosis timely.

Objective To test whether T helper 17 (Th17) cells was involved in the pathogenesis of multiple sclerosis (MS) by conducting a meta-analysis of the researches documenting the levels of Th17 cells and Th17-related cytokines [interleukin (IL)-17,IL-23] in patients with MS.Methods We identified articles reporting proportion of Th17 cells and the serum levels of IL-17,IL-23 in MS patients by searching CNKI,Wanfang med online,Embase,PubMed,Cochrane,WebofKnowledge,Food and Drug Administration (FDA).gov,and ClinicalTrials.gov.We registered this study at the International Prospective Register of Systematic Reviews (PROSPERO) (number CRD42017059113).The evidence level of selected studies was identified by guidelines from the Oxford Center for Evidence-Based Medicine 2011.We employed the Newcastle-Ottawa Quality Assessment Scale (Case Control Studies) to perform the included resear.Stata 12.0 was adopted for processing Meta-analysis of the data by using a random effects model.Results A total 16 researches were included in our Meta-analysis from 587 identified studies.The proportion of Th17 cells [1.89％(1.10％,2.69％),P＜0.01] and the levels of serum IL-17 [2.77(1.77,3.77) pg/ml,P＜0.01] and IL-23 [2.96(1.51,4.40)pg/ml,P＜0.01] increased dramatically in MS individuals compared with control subjects.Conclusion The results of this Meta-analysis has demonstrated that MS patients have a higher proportion of Th 17 cells in higher levels of serum IL-17 and IL-23 compared to control subjects,which has demonstrated that Th17 cells may play an vital role in the pathogenesis process of MS patients.

Objective To explore the efficacy and clinical value of texture analysis of conventional MRI image for the preoperative classification of meningiomas. Methods The MR images of 101 meningioma cases were retrospectively analyzed, all of them underwent routine MRI before operation, and confirmed by surgery and pathology,of whom 81 cases in WHO Ⅰ,19 cases in Ⅱand 1 case in Ⅲ.The maximum diameter of the tumor in MRI images was outlined as ROI by the Image J software.The histograms and grey-level co-occurrence matrix(GLCM)were used to measure the texture parameters,such as maximum value,minimum value,standard deviation(SD),skewness,kurtosis,angular second moment,contrast,inverse different moment, entropy, correlation and so on, and compared with postoperative pathological grading results.The independent sample t test or the Mann-Whitney U test was used to compare the difference of the histogram and the gray level co-occurrence matrix parameter of the MRI images between low-grade and high-grade meningiomas. ROC curves for statistically significant parameters were used to confirm their efficacy in predicting the preoperative grade of meningiomas.Results (1)In histogram parameters,the SDs of T2WI,FLAIR,ADC(SDT2WI,SDFLAIR,SDADC)of low grade meningioma were 14.764(10.928,17.932),9.753 (7.385,13.618) and 12.171 (9.138,19.691), the high grade meningiomas were 19.252 (13.580,25.715), 13.568 (8.936,17.108) and 16.636 (13.166,21.498), respectively. The difference between the two groups was statistically significant(Z values were-2.863,-2.250,-2.247,P respectively 0.004,0.024,0.025).The skewness values (SkeADC, SkeFLAIR) of ADC sequence and FLAIR sequence of low-grade meningiomas were 1.377 ± 1.172 and-1.327 ± 0.930, respectively, and the high-grade meningiomas were 2.503 ± 1.613 and-0.827±0.834,respectively.The differences between the two groups were statistically significant (t values were-2.196,-3.129, P values were 0.002,0.030, respectively). (2) In the GLCM parameters, entropy (EntT1WI, EntT2WI, EntFLAIR, EntADC) of the enhanced T1WI, T2WI, FLAIR and ADC sequences of low grade meningiomas were 6.881(6.174,7.305),6.534±0.598,6.019±0.588,6.040±0.588,high-grade meningiomas were 7.079 (6.742,7.739), 7.014 ± 0.514,6.370 ± 0.703,6.576 ± 0.450, respectively. And the difference between the two groups was statistically significant (Z=-2.007, t values were-3.294,-2.327,-3.245, P values were 0.045,0.001,0.022,0.002,respectively).(3)The ROC curves of EntADCand SkeADCwere plotted, and the area under the curve(AUC)was 0.768 and 0.710,respectively.Combined EntADC,SkeADCand EntT2WI were the best for the classification of meningiomas,and AUC was 0.799.Conclusion Texture analysis can provide more quantification information, which can be more accurately distinguishing high grade meningiomas and low grade meningiomas before surgery.

Objective To investigate the levels of T helper cell 17 (Th17), Th17-related cytokines in-terleukin 17 (IL-17) and interleukin 23 (IL-23) and regulatory T cell (Treg) in relapsing remitting multiple sclerosis (RRMS). Methods In a case-control study, plasma was collected from RRMS patients (n=20) and healthy subjects as control group (n=20). The percentages of Th17 and Treg cells and the levels of IL-17 and IL-23 were tested. The levels of Th17, Treg, IL-17 and IL-23 of the two groups were compared. Patients were treated with methylprednisolone. The levels of Th17, Treg, IL-17 and IL-23 of multiple sclerosis (MS) patients b efore and after treatment were compared. Expanded disability status scale (EDSS) score and the number of Gd-enhancing lesions were evaluated in the case group. Statistical analysis was made by body mass index (IBM) statistical program for social sciences (SPSS) 17.0 software. Independent sample t test was conducted to compare the measurement data of the case group and the healthy control group, and enumeration data were compared by χ2 test; paired sample t test was performed to compare the data of the case group before and after treatment; Pearson correlation analysis was made forthe variables of the MS group before treatment. Results In the RRMS group, the percentage of Th17 cells in peripheral blood was significantly higher than the control group [(2.10±0.45)％vs (1.09±0.20)％](t=9.130, P<0.01), the levels of Th17-related cytokines IL-17 and IL-23 were remarkably higher than the control group (IL-17:t=19.843, P<0.01;IL-23:t=22.747, P<0.01), and the percentage of Treg cells was significantly lower than the control group [(1.33 ±0.30)％vs (2.52±0.30)％], (t=12.422, P<0.01). The levels of Th17 and IL-17 were positively associated with EDSS score (Th17: r=0.458, P<0.05; IL-17: r=0.480, P<0.05), there was no significant-correlation between the level of IL-23 and EDSS score (r=0.368, P>0.05), and Th17, IL-17 and IL-23 were positively correlated with the number of Gd-enhancing lesions (Th17: r=0.446, P<0.05; IL-17: r=0.544, P<0.05; IL-23: r=0.461, P<0.05). The levels of Th17, IL-17 and IL-23 in the RRMS group after the treatment with methylprednisolone were obviously decreased than before treatment (Th17: t=5.747, P<0.01; IL-17: t=9.967, P<0.01; IL-23: t=14.697, P<0.01), while that of Treg was apparently increased (t=10.050, P<0.01). Compared with the control group, the levels of Th17, IL-17 and IL-23 in the RRMS group after treatment were higher (Th17: t=6.889, P<0.01;IL-17:t=7.185, P<0.01;IL-23:t=13.284, P<0.01), and the percentage of Treg was lower (t=7.622, P<0.01). EDSS score of the RRMS group after treatment was remarkably decreased than before treatment(t=6.190, P<0.01), but the number of Gd-enhanced lesions after treatment was no significantiy changed (t=1.453, P>0.05). Conclusion Th17/Treg expression imbalance and Th17-related cytokines IL-17, IL-23 may participate in the pathological process of MS, and they might be therapeutic target for MS.

[摘 要] 嵌合抗原受体基因修饰T（CAR-T）细胞免疫治疗被认为是最有前景的肿瘤治疗方法之一，效应CAR-T细胞的数量是决定CAR-T细胞疗法治疗效果的关键因素。CAR-T细胞的体外扩增耗时耗力，回输体内后，CAR-T细胞大量耗竭且难以浸润实体瘤，导致能有效抑制实体瘤的CAR-T细胞数量大幅下降。目前，CAR-T细胞的扩增方法在提高扩增特异性和治疗安全性等方面均存在问题，为CAR-T细胞疗法的临床转化造成困难。近年来，新型免疫激动剂及其下游信号的发现为CAR-T细胞扩增方案提供了更多选择，免疫激动剂给药方式的更新迭代进一步提高了其在体内扩增CAR-T细胞的安全性。本文分析了目前扩增CAR-T细胞面临的挑战，系统阐述了近年来在体内外扩增CAR-T细胞的新策略，为CAR-T细胞疗法的疗效和产能优化提供了新思路。

Objective: To explore the practicability and reproducibility of judgment method and assessment indexes for the end point of double eyelid surgery using 4+ 1 photography in supine position. Methods: From 2017 October to 2018 October, 158 patients were included and randomly divided into 2 groups. Photos were taken by 4 + 1 photography in supine position of 79 patients, while other 79 patients in control group were evaluated by traditional observation. By 4+ 1 photography in supine position, the surgeon stood at the head side of the patient, taking photos with eyes movement: looking straight forward, looking up, looking downward and eye-closed. It was to observe the upper eyelid creases, upper and lower tissues of double eyelid creases, and upper and lower eyelid margo palpebrae. In addition, the surgeon looked from patient′s feet to observe the indexes such as upper margo palpebrae, to make a decision whether the surgery could finish. In control group, the surgeon observed the upper eyelid creases, upper and lower tissues of double eyelid creases and upper and lower eyelid margo palpebrae. The patient has to sit if necessary. Results: All 158 patients were performed double eyelid surgery successfully. Average times of valuating end point was 1.20 by 4+ 1 Photography in supine position, and 1.53 in control group. The operation time of 4+ 1 photography group is (151.65±21.58) s, and control group were (241.53±33.53) s. The satisfaction level was increased to 89.87% in 4+ 1 photography group, compared to 78.48% in control group. Conclusions The 4+ 1 Photography method is simple and easy to practice. The observation indexes are accurate and comprehensive. It is useful in determining the end point of double eyelid plasty in supine position.