[Differentiation of temporal lobe epilepsy and temporal plus epilepsy using radiomics nomogram based on MPRAGE images].

A total of 82 patients with temporal lobe epilepsy (TLE) and temporal plus epilepsy (TPE)admitted in Xuanwu Hospital from January 1, 2019, to January 1, 2021 were restrospectively analyzed, including 41 males and 41 females, aged 2 to 52 (24±10) years. The patients were randomly divided into the training set (58 cases) and test set (24 cases) by Python. FreeSurfer software was used to segment the cortex of the affected hemisphere, defining 33 regions of interest (ROIs), and radiomics features were extracted by Python. After selecting features using the filter-based feature selection method, a radiomics model was constructed with a logistic regression classifier, and radiomics scores were calculated. Combining clinical characteristics with radiomics scores, a nomogram model was constructed using R software, the predictive accuracy of the model was assessed with the concordance index (C-index), and the model's goodness-of-fit was tested with the Hosmer-Lemeshow method. The results showed statistically significant differences between TLE and TPE patients in disease duration, intracranial electrode implantation, and hippocampal sclerosis (both P<0.05). The accuracy of the radiomics model in the training set and the test set was 91.4% and 87.5%, respectively. The nomogram model uses C-index to predict accuracy. Hosmer-Lemeshow method was used to test the goodness of fit, with AUCs of 0.95 (95%CI: 0.853-0.991) in the training set and 0.84 (95%CI: 0.676-0.999) in the test set. The study indicates that the radiomics nomogram model based on MPRAGE sequences can effectively differentiate TLE from TPE, providing reference for the development of personalized treatment plans in clinical practice.

[Alterations in erythrocytic oligomeric alpha-synuclein in patients with Parkinson's disease and multiple system atrophy].

Objective: To analyze the content of α-synuclein oligomer(O-α-Syn) in erythrocytes in patients with Parkinson's disease (PD) and multiple system atrophy (MSA) and the correlation with clinical symptoms. Methods: Two hundred and ninety-six PD patients and 85 MSA patients were recruited from the Department of Functional Neurosurgery and Neurology of Xuanwu Hospital, Capital Medical University from July 2020 to October 2021. Four hundred and three healthy controls (HC) were recruited from the Beijing Longitudinal Study of Aging community cohort during the same period. The levels of RBC-O-α-Syn were measured by enzyme-linked immunosorbent assay (ELISA). Univariate linear regression model was used to analyze the correlation between the content of RBD-O-α-Syn and various motor and non-motor functional scores, such as Unified Parkinson Disease Rating Scale (UPDRS) Ⅲ, Unified Multiple System Atrophy Rating Scale (UMSARS) Ⅲ, Mini-Mental State Examination (MMSE), rapid eye movement sleep disorder questionnaire-HongKong(RBDQ-HK) and Montreal Cognitive Assessment (MoCA). Receiver operating characteristic (ROC) curves was used to evaluate the specificity, sensitivity, and the area under the curve (AUC) of RBC-O-α-Syn in distinguishing PD and MSA patients from HC subjects. Results: The average age of HC subjects was (70±8) years old, the average age of PD patients was (64±9) years old, including 115 (38.9%) cases with tremor dominant PD (TD-PD), 132 cases (44.6%) of postural instability disorder predominant PD (PIGD-PD), and 142 cases (48.0%) of patients with H-Y stage 2. UPDRS Ⅲ score was 31.2±17.8. The mean age of MSA patients was (64±9) years, with the mean UMSARS Ⅱ score of 18.9±10.3. The non-motor symptoms of PD and MSA patients were significantly different from those of HC subjects (P<0.001). The levels of RBC-O-α-Syn in PD [(50±17) ng/mg] and MSA [(52±19) ng/mg] were significantly higher than those in HC subjects [(21±10) ng/mg] (P<0.001). The sensitivity and specificity of RBC-O-α-Syn in distinguishing PD patients and HC subjects were 87.16% (95%CI: 82.87%-90.50%) and 86.10% (95%CI: 82.38%-89.14%), with an AUC of 0.933 (95%CI: 0.914-0.951), and the sensitivity and specificity in distinguishing MSA patients and HC subjects were 85.88% (95%CI: 76.93%-91.74%) and 81.39% (95%CI: 77.30%-84.89%), with an AUC of 0.921 (95%CI: 0.884-0.957). The levels of RBC-O-α-Syn in PD patients with rapid eye movement sleep behavior disorder (RBD) were higher than that in PD patients without RBD [(53±16) ng/mg vs (48±17) ng/mg, P=0.029].The content of RBC-O-α-Syn in female PD patients and HC subjects was higher than that in male, but there was no significant difference between subjects of different ages and disease duration (P>0.05). In addition, RBC-O-α-Syn content was positively correlated with UPDRS Ⅲ (r=0.18, P=0.002) and the score of rapid eye movement sleep behavior disorder questionnaire(Hong Kong) (RBDQ-HK)(r=0.19, P<0.001). But there was no correlation with H-Y stage, non-motor symptoms scale (NMSS), MMSE, Moca, Hamilton Depression Scale (HAMD), Hamilton Anxiety Scale (HAMA) scores (all P>0.05). There was no correlation between RBC-O-α-Syn content and UMSARS Ⅱ, NMSS, MMSE, MoCA, HAMD, HAMA in patients with MSA (all P>0.05). Conclusions: Levels of RBC-O-α-Syn are significantly increased in PD and MSA patients. There are positive correlations between levels of RBC-O-α-Syn and scores of UPDRS Ⅲ and RBDQ-HK.

[The surveillance analysis of the adverse events following immunization of the domestic 13-valent pneumococcal polysaccharide conjugate vaccine in Zhejiang Province].

To evaluate the safety of the domestic 13-valent pneumococcal polysaccharide conjugate vaccine-tetanus toxoid protein (PCV13-TT) after its licensure. The adverse event following immunization (AEFI) and the vaccination data of PCV13-TT in Zhejiang province from July 2020 to October 2021 were collected from national adverse event following immunization surveillance system and Zhejiang provincial immunization information system. Descriptive epidemiological method was used for this analysis. From July 2020 to October 2021, 302 317 doses of PCV13-TT were administered in children under 6 years old in Zhejiang Province and 636 AEFI case reports were received, with a reporting rate of 21.04 per 10 000 doses. Of these AEFI cases, 97.17% were mild vaccine product-related reaction (20.54 per 10 000 doses) and 95.44% occurred in the 0-1 d after vaccination (20.08 per 10 000 doses). The most common clinical diagnoses of AEFI included fever (224 cases), redness (204 cases), and induration (190 cases), while allergic rash (11 cases) was the most common diagnosis among the abnormal reactions. In conclusion,the present results bolstered that the domestic PCV13-TT was generally well tolerated in children under 6 years old in Zhejiang Province.

[Examination and discriminant analysis of corneal biomechanics with CorVis ST in keratoconus and subclinical keratoconus].

OBJECTIVE: To compare the corneal biomechanical properties among keratoconus, subclinical keratoconus and normal corneas by using CorVis ST, and to estimate the effect of these biomechanical indices in discriminating keratoconus and subclinical keratoconus from normal. METHODS: A total of 76 eyes of 67 subjects were enrolled and divided into three groups. Keratoconus group included 24 eyes from 17 patients, subclinical keratoconus group included 12 eyes from 12 patients and normal group included 40 normal eyes from 40 subjects．All the eyes were assessed with CorVis ST and ten biomechanical parameters, intraocular pressure (IOP) and central corneal thickness (CCT) were obtained from this machine. The discrimination of biomechanical characteristic of the three groups based on the all indices was reflected by discriminant analysis and the Fisher discriminant function was established. RESULTS: The values of corneal biomechanics of keratoconus, subclinical keratoconus, normal eyes were increased in sequence, except for three indices: the second applamation time (A2T), time taken to reach highest concavity (HCT) and maximum corneal velocity during the first applanation (Vin). Three sets of data were among a statistically significant difference (P<0.05). There were statistically significant differences (P<0.05) between any two groups by comparing with such two indices: radius value of central concave curvature at highest concavity (HCR) and CCT. The grades of the three groups were obvious, evaluated by the discriminant function. The accuracy of reevaluation was 85% by validation method. The biggest contribution of indices in discriminant function was given by such four indices in sequence: CCT, HCR, maximum deformation amplitude of highest concavity (HCDA) and maximum corneal velocity during the second applanation (Vout). CONCLUSION: The corneal biomechanical properties of keratoconus and subclinical keratoconus were decreased compared with normal eyes. The biomechanical parameters based on CorVis ST showed a good performance for discriminating among keratoconus, subclinical keratoconus and normal corneas.

[Effect of different genotypes of PNPLA3 I148M on hepatocyte proliferation].

Objective: To explore the influence of patatin-like phospholipase domain containing-3 (PNPLA3) wild type 148I/I and mutant type 148M/M on HepG2 cell proliferation and the relative mechanisms. Methods: HepG2 cell line stably overexpressing PNPLA3 148I/I, 148M/M and negative control (NC) were set up. Cell counting kit-8 (CCK8) assay was used to measure cell viability. Edu assay was used to determine the ability of cell proliferation. Western blot was used to detect the protein levels in the phosphatidylinositol 3-kinases (PI3K) pathway. Enzyme-linked immunosorbent assay (ELISA) was used to detect proliferation-related PNPLA3 metabolites[arachidonic acid (AA) and lysophosphatidic acid (LPA)]. Quantitative real-time PCR was used to detect the expression level of prostaglandin G/H synthase 2 (PTGS2) and proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) associated with PNPLA3. Results: The cell viability of overexpression of PNPLA3 148M/M group was about 1/3 times higher than that of overexpression of PNPLA3 148I/I group, and the difference was statistically significant[(98.02±1.29)% vs (71.51±2.89)%, P<0.001]. There was no significant difference between overexpression of PNPLA3 148M/M group and negative control group[(98.02±1.29)% vs (100±2.61)%, P=0.181]. The proliferative activity of overexpression of PNPLA3 148M/M group was about 1/3 times higher than that of overexpression of PNPLA3 148I/I group, and the difference was statistically significant(46.46±1.83 vs 35.96±2.65, P=0.001). There was no significant difference between overexpression of PNPLA3 148M/M group and negative control group(46.46±1.83 vs 46.64±7.33, P=0.965). The PGC1α mRNA expression, total PI3K, PThr-308AKT, PSer2448-mammalian target of rapamycin (PSer2448-mTOR) and PGC1α protein expression levels in the overexpression of PNPLA3 148M/M group were higher than those in the overexpression of PNPLA3 148I/I group, but there were no significant differences in AA and LPA levels, as well as PTGS2 mRNA expression levels. Conclusion: PNPLA3 148M/M cell proliferation was stronger than PNPLA3 148I/I.

[Research progress of intense pulsed light treatment on meibomian gland dysfunction and relevant dry eye diseases].

Intense pulsed light (IPL) is a broad spectrum incoherent light which is produced by high-output xenon lamp. Since the invention of the first-generation IPL in 1994, IPL technology has been developing rapidly and extensively utilized in multiple fields relevant to dermatology across the world. In 2004, the fourth-generation IPL system was introduced with the optimal pulse technology (OPT) and has soon been used for cosmetic purposes all over the world. In 2002, Dr. Toyos found that the meibomian gland dysfunction (MGD) and dye eye disease (DED)symptoms of the rosacea patients who received IPL treatment have been improving significantly, therefore he started to explore the application of IPL system to treatment of dry eye disease. Several recent clinical studies have demonstrated the therapeutic potential of IPL for improving the symptoms and signs of MGD and DED. However, the published data of IPL treatment for MGD and DED is limited, the mechanism of IPL treatment for MGD and DED remained unclear and more relevant researches needed to be done in the future. This article discusses the clinical application history and general mechanism of IPL, and introduces the treatment of IPL for MGD and DED. (Chin J Ophthalmol, 2018, 54: 140-143).

[The association between corneal biomechanical parameters and visual field progression in patients with normal tension glaucoma].

Objective: To evaluate the association between corneal biomechanical parameters and visual field (VF) progression in normal tension glaucoma (NTG) using the Corvis-ST device, and to evaluate the ability of corneal biomechanical parameters to predict the VF progression. Methods: Corneal biomechanical parameters of newly diagnosed NTG patients were obtained using Corvis-ST in the baseline follow-up visit. The VF progression was defined as a 4-point increase in the Advanced Glaucoma Intervention Study (AGIS) score compared to the baseline in three consecutive follow-up visits (per 3-6 months). Corneal biomechanical parameters were compared between progressive and nonprogressive VF loss eyes using the independent-sample t test and Mann-Whitney U test. Spearman correlation analysis was used to explore the relationship between the corneal biomechanical parameters and the VF progression. Receiver operating characteristic curves were studied for the parameters and the sensitivity and specificity for distinguishing between progressive and nonprogressive glaucomatous eyes. The areas under the receiver operating characteristic curves (AUC) were also evaluated. Results: Sixty patients with NTG were enrolled in this study. Among them, 12 were lost to follow-up. A total of 48 patients completed all follow-up visits on schedule. Eleven of them were excluded due to one or more uncontrolled intraocular pressure (IOP) during the follow-up (less than 30% IOP reduction from the baseline). Thirty-seven eyes of 37 diagnosed NTG patients were enrolled. Ten eyes reached a progression endpoint. There was no significant difference in age, central corneal thickness, axial length, baseline IOP or baseline VF between the two groups. There was significant difference in Time A1 [(7.10±0.17) ms vs. (7.37±0.28) ms, t=-3.357, P=0.002], Length A1 [1.74(1.61, 1.77) mm vs. 1.78(1.77, 1.79) mm, Z=-3.036, P=0.002], Velocity A1 [0.16(0.14, 0.16) m/s vs. 0.15(0.14, 0.15) m/s, Z=-2.627, P=0.009] and DefAmpl HC [(1.22±0.13) mm vs. (1.12±0.11) mm, t=2.601, P=0.013] between progressive and nonprogressive glaucomatous eyes. Correlation analysis showed that Time A1, Length A1, Velocity A1 and DefAmpl HC were correlated with VF progression (r=-0.521, -0.463, 0.401, 0.349, P<0.05) . Time A1 demonstrated the highest AUC (0.817, P=0.001), followed by Length A1 (0.780, P=0.003), Velocity A1 (0.734, P=0.012) and DefAmpl HC (0.713, P=0.022). The cut-off set of Time A1 was 7.2 ms, the sensitivity was 80.0%, and the specificity was 82.8%. Conclusions: There were differences in corneal biomechanical parameters between eyes with progressive and nonprogressive VF loss in patients with NTG. There were lower Time A1 and Length A1 values and higher Velocity A1 and DefAmpl HC values in progressive glaucomatous eyes. This indicates a quicker response to reach first degree applanation and a larger degree of corneal deformability in progressive eyes. It is predicted that the easier deforming of the cornea, the smaller tolerance of the sclera and lamina cribros on IOP, making the optic disc more vulnerably. This may be one of the causes of glaucomatous optic nerve damage. Time A1 was the best parameter to predict the progression of VF among the corneal biomechanical parameters obtained by Corvis-ST. (Chin J Ophthalmol, 2018, 54: 171-176).

[Association between SORCS1 rs1416406 and therapeutic effect of exenatide].

Objective: To study the relationship between SORCS1 gene rs1416406 and efficiency of exenatide. Methods: Between August 2010 and August 2012, a hundred and one newly diagnosed patients with type 2 diabetes mellitus (T2DM) were from CONFIDENCE study covering 25 university-affiliated hospitals in 13 provinces of China. All patients received exenatide treatment for 48 weeks. Hemoglobin A1c (HbA1c), fasting plasma glucose (FPG), body mass index (BMI), oral glucose tolerance test (OGTT) glucose and insulin levels were measured before and after therapy. ß-cell function was assessed by fasting proinsulin/insulin (PI/I), disposition index (DI) and acute insulin response (AIR). SORCS1 gene rs1416406 was genotyped by improved multiple ligase detection reaction. The relationship between rs1416406 and the glucose-lowering effect as well as ß-cell function improvement of exenatide was analyzed by multiple linear regression. Results: There were statistically significant differences of HbA1c, FPG, 2 h plasma glucose (2 h PG), ß-cell function (PI/I, DI and AIR) and changes of PI/I in three genotypes (GG, GA, AA) of rs1416406 between baseline and 48-week therapy of exenatide (all P<0.05). No statistically significant difference was found in changes of HbA1c, FPG, 2 h PG, DI, AIR except for PI/I, after stratifying by genotypes of rs1416406. Multiple linear regression analysis showed rs1416406 was significantly associated with the PI/I change (P<0.05) after adjustment of age, sex, baseline BMI, HbA1c and PI/I. Conclusion: SORCS1 gene rs1416406 was associated with the PI/I improvement induced by exenatide. Patients carrying GG genotype had greater reduction in PI/I after exenatide treatment as compared with those carrying allele A. The results suggests that the newly diagnosed T2DM patients with GG genotype might obtain more benefit from the early treatment of exenatide .

[Evaluation of short-term effect of intense pulsed light combined with meibomian gland expression in the treatment of meibomian gland dysfunction].

Objective: To explore short-term effect of intense pulsed light (IPL) combined with meibomian gland expression in treating meibomian gland dysfunction (MGD). Methods: This study was a prospective, randomized, double-masked, controlled study. Forty-four MGD patients were enrolled in the study and received three consecutive IPL treatments with an interval of 4 weeks. One eye of each patient was randomly assigned as the study eye receiving the IPL therapy with an energy of 14-16 J/cm(2), and the fellow eye was as the control eye receiving a placebo therapy with 0 J/cm(2). Meibomian gland expression was immediately performed after the IPL treatment in both eyes. Efficacy was evaluated through assessment of the meibomian gland yielding secretion score (MGYSS) , SPEED questionnaire, tear film break-up time (TBUT), cornea fluorescein staining and infrared meibography. Safety was evaluated through best spectacle corrected visual acuity, intraocular pressure, slit lamp examination and fundus examination. These examinations were performed before and after each treatment. Results: Significant improvements were observed in the MGYSS and TBUT after IPL treatments (P<0.05). The improvements compared to the baseline of MGYSS at the upper eyelid in the treatment eyes were significantly higher than those in the control eyes after the first treatment (Z=-2.036, P=0.003). The improvements compared to baseline of MGYSS at the lower eyelid and the TBUT in the treatment eyes were significantly higher than those in the control eyes after the second treatment (Z=-2.999 and -2.036, respectively P=0.007 and 0.042, respectively). SPEED and cornea fluorescein staining were decreased in both eyes after IPL treatments, but there was no statistical difference between the two eyes. No obvious complication was observed in the study. Conclusions: IPL treatment combined with meibomian gland expression is an efficient and safe therapy, and can increase meibomian gland yielding secretion, increase the TBUT, relieve the symptoms and repair the corneal epithelium defects for MGD eyes. (Chin J Ophthalmol, 2017, 53: 675-681).

Effect of MSTN propeptide protein on the growth and development of Altay lamb muscle.

Prokaryotic expression technology was used to express maltose-binding protein binding myostatin (MSTN) propeptide fusion protein. Six disease-free Altay lambs were used in this study. The right leg gastrocnemii were injected with MSTN recombinant propeptide protein. The left leg gastrocnemii (the control group) were injected with the same dose of phosphate based saline. The lambs were fed during four months under the same conditions and then slaughtered. Gastrocnemius samples were hematoxylin-eosin stained and the size of the muscle fibers was measured. A real-time polymerase chain reaction (RT-PCR) showed that single gastrocnemius cells in the experimental group had an average area of 1163.01 µm(2), while it was 845.09 µm(2) in the control group (P < 0.05). This indicates that the MSTN propeptide biological agents had an inhibitory effect on MSTN. In order to reveal its mechanism, RT-PCR was conducted to detect the expression of the differentiation-associated genes MyoD, Myf5, Myogenin, p21, and Smad3. The results showed that, in the MSTN propeptide biological agent injected group, expression levels of MSTN, Smad3, and p21 were lower than the control group, while Myf5, MyoD, and Myogenin were higher compared to the control group. This indicates that, when expression of the MSTN gene was inhibited, muscle cell differentiation and growth can be promoted by Smad3 up-regulated expression of Myf5, MyoD, and Myogenin.

[The effect of long-term contact lens wear on the morphology of meibomian glands].

OBJECTIVE: To evaluate the effect of long-term contact lens (CL) wear on the morphology of meibomian glands (MGs) using meiboscore and digital analysis. METHODS: Retrospective study. Sixty right eyes of sixty patients were involved in this study, and the data were analyzed retrospectively. According to the duration of CL wear, all patients were divided into three groups, nonwear group (n=21), short-term group (duration of CL wear ≤3 years, n=19) and long-term group (duration of CL wear>3 years, n=20). Digital images of MGs obtained by meibography were analyzed using Image J software, providing the area percentage of MGs loss. The meiboscores were also examined, and the data were analyzed using one-way ANOVA and Kruskal-Wallis test. RESULTS: Ten out of 21 nonwearers were scored 0 point, and 11 were scored 1 point in the upper lid meiboscores, while 7 were scored 0 point, 9 were scored 1 point, and 5 were scored 2 points in the total meiboscores. Seven out of 19 short-term wearers were scored 0 point, 10 were scored 1 point, and 2 were scored 2 points in the upper lid meiboscores, while 5 were scored 0 point, 6 were scored 1 point, 6 were scored 2 points, and 2 were scored 3 points in the total meiboscores. Four out of 20 long-term wearers were scored 0 point, 7 were scored 1 point, and 9 were scored 2 points in the upper lid meiboscores, while 3 were scored 0 point, 4 were scored 1 point, 4 were scored 2 points, 4 were scored 3 points, 4 were scored 4 points, and 1 was scored 5 points in the total meiboscores. The meiboscores of the upper eyelid and total meiboscores among the three groups were significantly different (Hc=9.967, P=0.007; Hc=9.725, P=0.008). The meiboscores of the upper eyelid and total meiboscores were significantly higher in the long-term group compared to the nonwear group (Z=102.500, P=0.003, Z=100.500, P=0.003) and the short-term group (Z=120.500, P=0.050, Z=117.500, P=0.041). No significant difference was found between the short-term group and the nonwear group. The median of the MGs loss area percentage in the upper eyelid of the nonwear, short-term and long-term groups was 9.2%, 13.3% and 16.7%, respectively. The median of the total MGs loss area percentage in the nonwear, short-term and long-term groups were 6.6%, 8.8% and 13.0%, respectively. The above medians were significantly different among the three groups (Hc=6.390, P=0.041; Hc=7.019, P=0.030). They were significantly larger in the long-term wearers than the nonwearers (Z=120.500, P=0.019, Z=120.500, P=0.009). No significant difference was found between the short-term group and the nonwear group, or between the short-term group and the long-term group. No significant differences in the meiboscores or MGs loss area percentage in the lower eyelid were noticed among the three groups. The area under the curve of total area percentage of MGs loss in receiver operating characteristic analysis was 0.981 (P<0.001). CONCLUSIONS: Long-term (more than 3 years) CL wear can cause MGs loss. Digital analysis is helpful in the morphologic evaluation of MGs. (Chin J Ophthalmol, 2016, 52: 604-609).

Genotype I of hepatitis B virus was found in east Xishuangbanna, China and molecular dynamics of HBV/I.

There is a dearth of data about the prevalence of hepatitis B virus (HBV) infection in Mengla, China; and no detailed analysis of the molecular evolution of genotype I in Asia. In this study, 909 serum samples from ethnic minority people in China were obtained. Serological assay and HBV S-gene amplification were carried out, and phylogenetic and evolutionary dynamics analysis of 62 HBV/I S-gene was performed. On this survey, 153 individuals were tested HBsAg-positive. Genotypes of S-gene were classified into three groups: C, B and I. Under the strict model and the relax model, the estimated evolutionary rates for HBV/I were 3.74 × 10(-4) and 6.93 × 10(-4) substitution/site/year, respectively. However, when the geographic origin was taken into account, the mean substitution rates were increased. Estimated time to most recent ancestor of genotype I varied from ~30 to ~70 years ago. The Bayesian sky plot showed a rapid spread of HBV/I at the end of 1980s. Peculiar nucleotides distributed were observed in the subgenotype I1/I2. In conclusion, higher prevalence of HBV infection was observed in Mengla county. Multifactors like timescale and spatial locations should be integrated to provide a better interpretation of the HBV/I evolutionary history in the region.

Expression of recombinant myostatin propeptide pPIC9K-Msp plasmid in Pichia pastoris.

Myostatin propeptide can inhibit the biological activity of myostatin protein and promote muscle growth. To express myostatin propeptide in vitro with a higher biological activity, we performed codon optimization on the sheep myostatin propeptide gene sequence, and mutated aspartic acid-76 to alanine based on the codon usage bias of Pichia pastoris and the enhanced biological activity of myostatin propeptide mutant. Modified myostatin propeptide gene was cloned into the pPIC9K plasmid to form the recombinant plasmid pPIC9K-Msp. Recombinant plasmid pPIC9K-Msp was transformed into Pichia pastoris GS115 by electrotransformation. Transformed cells were screened, and methanol was used to induce expression. SDS-PAGE and western blotting were used to verify the successful expression of myostatin propeptide with biological activity in Pichia pastoris, providing the basis for characterization of this protein.

[Prediction of superantigen active sites and clonal expression of staphylococcal enterotoxin-like W].

Objective: The docking and superantigen activity sites of staphylococcal enterotoxin-like W (SElW) and T cell receptor (TCR) were predicted, and its SElW was cloned, expressed and purified. Methods: AlphaFold was used to predict the 3D structure of SElW protein monomers, and the protein models were evaluated with the help of the SAVES online server from ERRAT, Ramachandran plot, and Verify_3D. The ZDOCK server simulates the docking conformation of SElW and TCR, and the amino acid sequences of SElW and other serotype enterotoxins were aligned. The primers were designed to amplify selw, and the fragment was recombined into the pMD18-T vector and sequenced. Then recombinant plasmid pMD18-T was digested with BamHⅠand Hind Ⅲ. The target fragment was recombined into the expression plasmid pET-28a(+). After identification of the recombinant plasmid, the protein expression was induced by isopropyl-beta-D- thiogalactopyranoside. The SElW expressed in the supernatant was purified by affinity chromatography and quantified by the BCA method. Results: The predicted three-dimensional structure showed that the SElW protein was composed of two domains, the amino-terminal and the carboxy-terminal. The amino-terminal domain was composed of 3 α-helices and 6 ß-sheets, and the carboxy-terminal domain included 2 α-helices and 7 antiparallel ß-sheets composition. The overall quality factor score of the SElW protein model was 98.08, with 93.24% of the amino acids having a Verify_3D score ≥0.2 and no amino acids located in disallowed regions. The docking conformation with the highest score (1 521.328) was selected as the analysis object, and the 19 hydrogen bonds between the corresponding amino acid residues of SElW and TCR were analyzed by PyMOL. Combined with sequence alignment and the published data, this study predicted and found five important superantigen active sites, namely Y18, N19, W55, C88, and C98. The highly purified soluble recombinant protein SElW was obtained with cloning, expression, and protein purification. Conclusions: The study found five superantigen active sites in SElW protein that need special attention and successfully constructed and expressed the SElW protein, which laid the foundation for further exploration of the immune recognition mechanism of SElW.

[Population structure of food-borne Staphylococcus aureus in China].

Objective: To understand the population structure of food-borne Staphylococcus (S.) aureus in China. Methods: Whole genome sequencing was used to analyze 763 food-borne S. aureus strains from 16 provinces in China from 2006 to 2020. Multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and staphylococcal chromosome cassettemec (SCCmec) typing were conducted, and minimum spanning tree based on ST types (STs) was constructed by BioNumerics 7.5 software. Thirty-one S. aureus strains isolated from imported food products were also included in constructing the genome phylogenetic tree. Results: A total of 90 STs (20 novel types) and 160 spa types were detected in the 763 S. aureus isolates. The 72 STs (72/90, 80.0%) were related to 22 clone complexes. The predominant clone complexes were CC7, CC1, CC5, CC398, CC188, CC59, CC6, CC88, CC15, and CC25, accounting for 82.44% (629/763) of the total. The STs and spa types in the predominant clone complexes changed over the years. The methicillin-resistant S. aureus (MRSA) detection rate was 7.60%, and 7 SCCmec types were identified. The ST59-t437-Ⅳa (17.24%, 10/58), ST239-t030-Ⅲ (12.07%, 7/58), ST59-t437-Ⅴb (8.62%, 5/58), ST338-t437-Ⅴb (6.90%, 4/58) and ST338-t441-Ⅴb (6.90%, 4/58) were the main types in MRSA strains. The genome phylogenetic tree had two clades, and the strains with the same CC, ST, and spa types clustered together. All CC7 methicillin sensitive S. aureus strains were included in Clade1, while 21 clone complexes and all MRSA strains were in Clade2. The MRSA strains clustered according to the SCCmec and STs. The strains from imported food products in CC398, CC7, CC30, CC12, and CC188 had far distances from Chinese strains in the tree. Conclusions: In this study, the predominant clone complexes of food-borne strains were CC7, CC1, CC5, CC398, CC188, CC59, CC6, CC88, CC15, and CC25, which overlapped with the previously reported clone complexes of hospital and community-associated strains in China, suggesting that close attention needs to be paid to food, a vehicle of pathogen transmission in community and food poisoning.

[Cloning and expression of recombinant truncated SElX protein and evaluation on the related emetic activities].

Objective: To analyze the amino acid polymorphism of truncated Staphylococcal enterotoxin-like toxin X (tSElX), and to evaluate its related emetic activities. Methods: Sequence of tselx was compared with both the genome sequence of 145 CC398 strains completed in our research group and the NCBI database. Primers were designed to amplify the target gene of tselx, and the fragment was recombined into pMD18-T vector and sequenced. PCR product was digested with BamHⅠ and EcoRⅠ, and constructed into plasmid pGEX-6P-1 and pET-28a (+). After recombinant plasmid was identified, the protein expression was induced by IPTG. Proteins expressed in the form of inclusion bodies were denatured and renatured, then purified by affinity chromatography and ultrafiltration. Purified tSElX protein was then fed to common marmosets with the dose of 250 µg/kg to observe the vomiting reaction. Results: tselx gene was present in 145 strains of CC398 strains from the different origins (patients, healthy people and animals) in China. Homology of the amino acid sequence of the protein from the Chinese strains appeared 100.0%, while the homology with the four American strains were 97.8%(1) and 98.9%(3), respectively. Through two sets of expression systems and different induction conditions, tSElX was expressed in the form of inclusion bodies. The high purity soluble recombinant tSElX was thus obtained by denaturated and renaturated processes. At the dose of 250 µg/kg, tSElX protein did not cause vomiting in common marmosets. Conclusions: Results of this study showed that the amino acid sequence of tSElX was highly conserved and was universally present in a particular clone group. We obtained soluble recombinant tSElX protein with high purity. We also noticed that tSElX did not have the animal emetic activity at a dose of 250 µg/kg.

[Expression, characterization of recombinant human soluble BAFF secreted from CHO cell].

B cell-activating factor of the TNF family (BAFF) is critical for B cell maturation and survival. Here, we constructed a stable CHO cell line, in which the expression level of soluble form of BAFF (sBAFF) was raised from 0.13 microg/ml to 0.55 microg/ml. Purified recombinant sBAFF from these CHO cells not only bound to its receptors but also co-stimulated the proliferation of human peripheral blood B lymphocyte in vitro. These results provided us with a useful basis for further studies about sBAFF-related research.

Adsorption and desorption of atrazine on carbon nanotubes.

The potential impact of carbon nanotubes (CNTs) on human health and the environment is receiving more and more attention. The high surface area of CNTs tends to adsorb a large variety of toxic chemicals, which may enhance the toxicity of CNTs and/or toxic chemicals. In this study, adsorption and desorption of atrazine on carbon nanotubes from aqueous solution were studied through batch reactors. The adsorption equilibrium isotherms were nonlinear and were fitted by Freundlich, Langmuir, and Polanyi-Manes models. It was found that the Polanyi-Manes model described the adsorption process better than other two isotherm models. Together with the "characteristic curve," the Polanyi adsorption potential theory is applicable to describe the adsorption process of atrazine on CNTs. Thermodynamic calculations indicated that the adsorption reaction of atrazine on CNTs is spontaneous and exothermic. The desorption data showed that no significant desorption hysteresis occurred. High adsorption capacity and adsorption reversibility of atrazine on CNTs suggest that CNTs have high health and environmental risks, whereas they have potential applications for atrazine removal from water.

Relationship between Hashimoto's thyroiditis and papillary thyroid carcinoma in children and adolescents.

OBJECTIVE: We investigated the relationship between Hashimoto's thyroiditis (HT) and papillary thyroid carcinoma (PTC) in children and adolescents. PATIENTS AND METHODS: We carried out a retrospective study of thyroidectomies performed from 2004 to 2017 at The First People's Hospital and the Tumor Hospital of Yunnan Province (Kunming, China). The occurrence and features of PTC and benign thyroid disease (BTD) in children and adolescents (age ≤ 20 years) were compared. RESULTS: We evaluated 258 consecutive thyroidectomies. Among children and adolescents with PTC, 23 cases were histopathologically confirmed as HT. Mean tumor diameter was smaller in children and adolescents with PTC than in those with BTD. Thyroid-stimulating hormone (TSH) level was abnormally elevated in a greater proportion of children and adolescents with PTC as compared to those with BTD or youths with PTC. The proportion of thyroglobulin antibody (TGAb)- and thyroid peroxidase antibody (TpoAb)-positive children and adolescents was higher in the PTC than in the BTD group. Among children and adolescents with PTC, 23 had HT as compared to two in the BTD group. The proportion of children/adolescents with abnormally elevated TSH levels was higher for the PTC combined with HT group than for the PTC without HT group. A multivariate conditional logistic regression analysis showed that elevated TGAb was an independent risk factor for PTC in children and adolescents. CONCLUSIONS: HT is associated with an increased occurrence of PTC in children and adolescents.