OBJECTIVESince glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.

METHODSAccording to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.

RESULTSIn the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).

CONCLUSIONSRT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.

Adolescent ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Glucosephosphate Dehydrogenase Deficiency ; diagnosis ; genetics ; Humans ; Infant ; Male ; Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To explore the endoplasmic reticulum stress (ERS) apoptosis pathway in unilateral ureteral obstruction (UUO) mechanism of renal interstitial fibrosis in rats.Methods The rats were randomly divided into con-trol group and model group,UUO group line ligation of the left ureter,three days 3,7 and 14 days HE and Masson staining were used to observe the renal pathological changes;Take blood retinal venous plexus,separat and determina-tion of serum blood urea nitrogen and creatinine;Western blot was used to find protein 78 (GRP78) glucose regula-tion,endoplasmic reticulum source sex transcription factor (CHOP) and apoptosis related proteins cysteine aspartic acid proteinase 3(caspase-3) and caspase-12 protein expression. Results Compared with control group,the visible UUO model group 1) expansion of renal tubule and renal interstitial fibrosis degree with the extension of ureteral ob-struction time and progressive increase;2) GRP78,CHOP,caspase-3 and caspase-12 protein expression in postoper-ative 3 d have to rise,as the obstruction prolonged,the protein expression more significantly(P<0.01).Conclusions Endoplasmic reticulum stress related trademark protein in UUO rat renal interstitial fibrosis is the increase in expres-sion may promote the early renal interstitial fibrosis and continuous progress.

Objective To explore the endoplasmic reticulum stress (ERS) apoptosis pathway in unilateral ureteral obstruction (UUO) mechanism of renal interstitial fibrosis in rats.Methods The rats were randomly divided into con-trol group and model group,UUO group line ligation of the left ureter,three days 3,7 and 14 days HE and Masson staining were used to observe the renal pathological changes;Take blood retinal venous plexus,separat and determina-tion of serum blood urea nitrogen and creatinine;Western blot was used to find protein 78 (GRP78) glucose regula-tion,endoplasmic reticulum source sex transcription factor (CHOP) and apoptosis related proteins cysteine aspartic acid proteinase 3(caspase-3) and caspase-12 protein expression. Results Compared with control group,the visible UUO model group 1) expansion of renal tubule and renal interstitial fibrosis degree with the extension of ureteral ob-struction time and progressive increase;2) GRP78,CHOP,caspase-3 and caspase-12 protein expression in postoper-ative 3 d have to rise,as the obstruction prolonged,the protein expression more significantly(P<0.01).Conclusions Endoplasmic reticulum stress related trademark protein in UUO rat renal interstitial fibrosis is the increase in expres-sion may promote the early renal interstitial fibrosis and continuous progress.

Objective To investigate the characteristics of muscle edema and fatty infiltration in thighs and relationship with clinical symptoms in Chinese patients with different phenotypes of dysferlinopathy.Methods A total of 32 patients were enrolled , including 13 limb-girdle muscular dystrophy 2B (LGMD2B), 13 Miyoshi myopathy (MM), 4 proximodistal myopathy and 2 hyper-creatine-kinase-emia.Clinical symptoms were evaluated using modified Gardner-Medwin and Walton ( GM-W) score.Muscle MRI was performed in thighs to observe fatty infiltration and edema.We then compared the age of onset , disease duration, GM-W score, muscle edema and muscle fatty infiltration between LGMD 2B and MM groups,and the relationship of muscle edema score and fatty infiltration score with disease duration and GM-W score in all patients.Results The median GM-W score was 4.00 (2.00,5.00) in all patients, 4.00 (3.00,4.50)in LGMD2B and 4.00(2.00,5.00)in MM, respectively.Muscle fatty infiltration appeared in 30 cases (93.75%), with the same pattern in LGMD2B and MM.The mean fatty infiltration score was 28.20 ±12.86 in all patients, 28.50 ±13.03 in LGMD2B and 29.00 ±12.63 in MM, respectively.Muscle edema appeared in 26 cases (81.25%) with same pattern in LGMB2B and MM.The mean edema score was 18.36 ±13.60 in all patients, 22.88 ±11.59 in LGMD2B and 16.77 ±13.80 in MM.The age of onset , disease duration, GM-W score, muscle fatty infiltration and edema score were not significantly different between LGMD2B and MM patients.Muscle fatty infiltration score significantly correlated with GM-W score (rs=0.737,P=0.000) and disease duration (rs=0.637,P=0.000).Conclusions Fatty infiltration and edema in thigh muscles are very common in patients with dysferlinopathy , with similar radiological changes in main subtypes.The muscle fatty infiltration can be used as a predictor of disease progression.

BACKGROUNDUnstable pelvic fractures are complex and serious injuries. Selection of a fixation method for these fractures remains a challenging problem for orthopedic surgeons. This study aimed to compare the stability of Tile C pelvic fractures fixed with two iliosacral (IS) screws and minimally invasive adjustable plate (MIAP) combined with one IS screw.

METHODSThis study was a biomechanical experiment. Six embalmed specimens of the adult pelvis were used. The soft tissue was removed from the specimens, and the spines from the fourth lumbar vertebra to the proximal one-third of both femurs were retained. The pubic symphysis, bilateral sacroiliac joints and ligaments, bilateral hip joints, bilateral sacrotuberous ligaments, and bilateral sacrospinous ligaments were intact. Tile C pelvic fractures were made on the specimens. The symphysis pubis was fixed with a plate, and the fracture on the posterior pelvic ring was fixed with two kinds of internal fixation in turn. The specimens were placed in a biomechanical machine at a standing neutral posture. A cyclic vertical load of up to 500 N was applied, and displacement was recorded. Shifts in the fracture gap were measured by a grating displacement sensor.

STATISTICAL ANALYSIS USEDPaired-samples t-test.

RESULTSUnder the vertical load of 100, 200, 300, 400, and 500 N, the average displacement of the specimens fixed with MIAP combined with one IS screw was 0.46, 0.735, 1.377, 1.823, and 2.215 mm, respectively, which was significantly lower than that of specimens fixed with two IS screws under corresponding load (P < 0.05). Under the vertical load of 500 N, the shift in the fracture gap of specimens fixed with MIAP combined with one IS screw was 0.261 ± 0.095 mm, and that of specimens fixed with two IS screws was 0.809 ± 0.170 mm. The difference was significant (P < 0.05).

CONCLUSIONThe stability of Tile C pelvic fractures fixed with MIAP combined with one IS screw was better than that fixed with two IS screws.

Adult ; Bone Screws ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; surgery

This study was aimed to investigate the effects of different stimulatory factors on proliferation and function of cytokine induced killer (CIK) cells. Peripheral blood mononuclear cells (PBMNC) were separated by Ficoll-Hypacue gradient. According to supplement of different stimulatory factors (CD28 mAb, IL-15 and IL-21), the experiment was divided into five groups:control group (CIK), CB28+IL-15+IL-21 group, IL-15+IL-21 group, CD28+IL-15 group and CD28+IL-21 group. Effects of different stimulatory factors on the proliferation of CIK cells were assayed by an automated hematology analyzer. Changes of granzyme B,perforin and CD107a were detected by flow cytometry. IL-10, IL-12, INF-γ and TNF-α were quantified by ELISA. Cytotoxicities on lung cancer cell line A549, breast adenocarcinoma cell line MFC-7 and human melanoma cell line HME1 were examined by lactate dehydrogenase release method. The results showed that there were significant differences among different groups. The highest proliferation index on days 10 was observed in group CD28mAb, IL-15 and IL-21(255.3 ± 6.3), which was higher than control group, IL-21+IL-15 group and CD28 mAb+IL-21 group (166.6 ± 13.5, 199.4 ± 15.0 and 228.8 ± 16.6) (P < 0.05). The expression of perforin in CD28 mAb+IL-15 group was higher than the other groups. The expression of perforin,GranB and CD107a of costimulatory groups was higher than control group. The cytotoxicities of CD28 mAb+IL-15 group on A549, MFC-7 and HME1 cells (82.2%, 59.3% and 70.6%) were much higher than that of control group (60.9%, 49.6% and 48.4%) (P < 0.05). The highest IFN-γsecretion was found in CD28 mAb, IL-15 and IL-21 groups. It is concluded that there are significant difference of proliferative capacity, cytokine secretion and cytotoxicity after being activated by different stimulatory factors. Adding corresponding stimulatory factors into the culture system displays a great value for target cells culture.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cytokine-Induced Killer Cells ; cytology ; drug effects ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-15 ; pharmacology ; Interleukins ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism

Objective:To test the validity and reliability of the Uighur version of Chinese Soldier Personality Questionnaire (CSPQ) in Uygur ethnic group of recruited youtt.Methods:Using the two-way checklist,all the items were judged by professors as their items belonging to test the content validity.Totally 101 Uygur population of permanent residents and totally 102 patients with schizophrenia in remission in sample 1 were tested for discrimination validity analysis.Totally 460 Uygur youths were recruited to complete the Uygur form of CSPQ for subscale normal distribution analysis and reliability analysis in sample 2.Totally 118 students of Urumqi College of Land Army from sample 3 were selected and retested for test-retest reliability with three weeks interval.Results:Uygur form of CSPQ had 283 items and 8 dimensions.Classification and recognition rate judged by professors ranged from 74.6％ to 91.5％.Patients with schizophrenia scored higher than normal people in all scales.Reliability coefficients of the 8 dimensions ranged from 0.69 to 0.91,and the test-retest reliability ranged from 0.85 to 0.92.Conclusion:It suggests that Uighur version of Chinese Soldier Personality Questionnaire is of good validity and reliability.

To observe metabolic abnormalities, histology changes and eNOS expression of aorta in type 2 diabetes rat.And to observe intervention effect of rosiglitazone.Methods 80 male Wistar rats were randomized to control group, high diet group, diabetes group, and rosiglitazone treatment group (diabetes plus rosiglitazone treatment).Type 2 diabetes models were developed and rosiglization group was treated with rosiglitazone .Six weeks and twelve weeks after treatment with rosiglitazone, blood glucose, endothlin and nitric oxide were tested.Histology changes of aorta in different groups were observed under microscopy .Meanwhile the protein and mRNA expression of eNOS in aorta were examined.Results 1)Compared with control group, ET in high fat diet group, diabetes group and rosiglitazone group increased significantly , and the level of NO decreased significantly at 6 week and 12 week.At 12 week, ET in diabetes group increased, and NO decreased significantly than that of high fat diet group and rosiglitazone group.2)Histology changes were observed in high fat diet group , diabetes group and rosiglitazone group at 12 week.3)Compared with control group, protein and mRNA expression in high fat diet group , diabetes group and rosiglitazone group were down regulated at 6 week and 12 week.And protein expression in diabetes group was down regulated than that in rosiglitazone group .Conclusions Rosiglization can ease the endothelial dysfunction in type 2 diabetes rat.

Objective:To investigate the value of CT features in predicting visceral pleural invasion (VPI) in clinical stage ⅠA peripheral lung adenocarcinoma under the pleura.Methods:The CT signs of 274 patients with clinical stage ⅠA peripheral lung adenocarcinoma under the pleura diagnosed in Changzheng Hospital of Naval Medical University from January 2015 to November 2021 were retrospectively analyzed. According to the ratio of 6∶4, 164 patients collected from January 2015 to August 2019 were used as the training group, and 110 patients collected from August 2019 to November 2021 were used as the validation group. The maximum diameter of the tumor (T), the maximum diameter of the consolidation part (C), and the minimum distance between the lesion and the pleura (DLP) were quantitatively measured, and the proportion of the consolidation part was calculated (C/T ratio, CTR). The CT signs of the tumor were analyzed, such as the relationship between the tumor and the pleura classification, the presence of a bridge tag sign, the location of the lesion, density type, shape, margin, boundary and so on. Variables with significant difference in the univariate analysis were entered into multivariate logistic regression analysis to explore predictors for VPI, and a binary logistic regression model was established. The predictive performance of the model was analyzed by receiver operating characteristic curve in the training and validation group.Results:There were 121 cases with VPI and 153 cases without VPI among the 274 patients with lung adenocarcinoma. There were 79 cases with VPI and 85 cases without VPI in the training group. Univariate analysis found that the maximum diameter of the consolidation part, CTR, density type, spiculation sign, vascular cluster sign, relationship of tumor and pleura and bridge tag sign between patients with VPI and those without VPI were significantly different in the training group( P<0.05). Multivariate logistic regression analysis found the relationship between tumor and pleura [taking type Ⅰ as reference, type Ⅱ (OR=6.662, 95%CI 2.364-18.571, P<0.001), type Ⅲ (OR=34.488, 95%CI 8.923-133.294, P<0.001)] and vascular cluster sign (OR=4.257, 95%CI 1.334-13.581, P=0.014) were independent risk factors for VPI in the training group. The sensitivity, specifcity, and area under curve (AUC) for the logistic model in the training group were 62.03%, 89.41% and 0.826, respectively, using the optimal cutoff value of 0.504. The validation group obtained an sensitivity, specifcity, and AUC of 92.86%, 47.06%, and 0.713, respectively, using the optimal cutoff value of 0.449. Conclusion:The relationship between the tumor and the pleura and the vascular cluster sign in the CT features can help to predict visceral pleural invasion in the clinical stage ⅠA peripheral lung adenocarcinoma under the pleura.

Objective:To understand the health self-assessment and related influencing factors in the migrant workers who received of physical examination in Shanghai， so as to provide reference for formulating health service policies for migrant workers. Methods:Taking Shanghai "Intelligent Blue Collar" Health Science Popularization Service Station as the investigation site， 8，100 urban workers were surveyed by questionnaire. The questionnaire includes basic information， health self-assessment， basic medical insurance coverage and satisfaction， subjective well-being and so on. Results:There were significant differences between migrant workers and local workers in health self-assessment， two-week disease prevalence rate， and chronic disease prevalence rate（χ²=32.981，P<0.05；χ²=4.554，P<0.05；χ²=86.695，P<0.05）. The differences between migrant workers and local workers were also significant in the basic medical insurance coverage rate and the level of satisfaction （χ²=221.592，P<0.05；χ²=30.950，P<0.05）. There was no significant difference in subjective well-being between these two groups（χ²=8.787，P>0.05）. The health self-assessment of older migrant workers was significantly better than that of new generation migrant workers（β=0.062，P<0.05）， and was significantly worse in female migrant workers than that in male migrant workers（β=-0.120，P<0.05）. The health self-assessment was significantly better in migrant workers with agricultural household registration than that in non-agricultural household registration（β=0.060，P<0.05）. The higher the satisfaction of basic medical insurance，the higher the subjective well-being and the better the health self-assessment（β=0.148，P<0.05；β=0.422，P<0.05）. Conclusion:The health status of migrant workers in Shanghai who received physical examination is better than that of local workers， and the participation rate of basic medical insurance is lower than that of local workers. Age， gender and household registration are the important factors influencing the health self-assessment. Satisfaction with basic medical insurance level and subjective well-being is a significant factor to predict health self-assessment positively . We should further improve the fairness of medical insurance benefits for migrant workers， improve subjective well-being， and ensure the health fairness of the group.