[Endometrioid adenocarcinoma with proliferated stromal cells, hyalinization and cord-like formations: A case report].

Corded and hyalinized endometrioid carcinoma (CHEC) is a morphologic variant of endo-metrioid adenocarcinoma. The tumor exhibits a biphasic appearance with areas of traditional low-grade adenocarcinoma merging directly with areas of diffuse growth composed of epithelioid or spindled tumor cells forming cords, small clusters, or dispersed single cells. It is crucial to distinguish CHEC from its morphological mimics, such as malignant mixed mullerian tumor (MMMT), because CHECs are usually low stage, and are associated with a good post-hysterectomy prognosis in most cases while the latter portends a poor prognosis. The patient reported in this article was a 54-year-old woman who presented with postmenopausal vaginal bleeding for 2 months. The ultrasound image showed a thickened uneven echo endometrium of approximately 12.2 mm and a detectable blood flow signal. Magnetic resonance imaging revealed an abnormal endometrial signal, considered endometrial carcinoma (Stage Ⅰ B). On hysterectomy specimen, there was an exophytic mass in the uterine cavity with myometrium infiltrating. Microscopically, most component of the tumor was well to moderately differentiated endometrioid carcinoma. Some oval and spindle stromal cells proliferated on the superficial surface of the tumor with a bundle or sheet like growth pattern. In the endometrial curettage specimen, the proliferation of these stromal cells was more obvious, and some of the surrounding stroma was hyalinized and chondromyxoid, which made the stromal cells form a cord-like arrangement. Immunostains were done and both the endometrioid carcinoma and the proliferating stroma cells showed loss of expression of DNA mismatch repair protein MLH1/PMS2 and wild-type p53 protein. Molecular testing demonstrated that this patient had a microsatellite unstable (MSI) endometrial carcinoma. The patient was followed up for 6 months, and there was no recurrence. We diagnosed this case as CHEC, a variant of endometrioid carcinoma, although this case did not show specific ß-catenin nuclear expression that was reported in previous researches. The striking low-grade biphasic appearance without TP53 mutation confirmed by immunohistochemistry and molecular testing supported the diagnosis of CHEC. This special morphology, which is usually distributed in the superficial part of the tumor, may result in differences between curettage and surgical specimens. Recent studies have documented an aggressive clinical course in a significant proportion of cases. More cases are needed to establish the clinical behaviors, pathologic features, and molecular profiles of CHECs. Recognition of the relevant characteristics is the prerequisite for pathologists to make correct diagnoses and acquire comprehensive interpretation.

[Effect of hyperandrogenism on pregnancy outcomes in women with polycystic ovary syndrome undergoing in vitro fertilization/intracytoplasmic sperm injection-embryo transfer].

Objective: To explore the effects of hyperandrogenism (HA) on pregnancy outcomes in women with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET). Methods: A retrospective study was conducted on infertile women with PCOS undergoing IVF/ICSI-ET from January 2017 to June 2021 in our center. Patients were divided into HA group and NON-HA group according to the levels of testosterone. Propensity score matching (PSM) was used to balance the influence of female age and IVF/ICSI-ET for patients with gonadotropin-releasing hormone (GnRH)antagonist protocol and GnRH agonist protocol, separately. After the PSM procedure, 191 cases in HA group and 382 cases in NON-HA group, were included. Hormone levels and pregnancy outcomes were compared in the two groups. Results: The female age was comparable in two groups [HA: (29.6±3.7) vs NON-HA: (29.5±3.6), P=0.665]. The basal luteinizing hormone [(10.82±6.73) vs (7.76±5.30) IU/L], testosterone [(3.27±0.97) vs (1.60±0.59) nmol/L], free androgen index (7.13 vs 2.77), anti-mullerian hormone [(11.37±5.74) vs (9.67±4.67) ng/ml], fasting glucose [(5.18±0.49) vs (5.06±0.42) mmol/L], 1h glucose [(9.34±2.42) vs (7.99±2.21) nmol/L], 2 h glucose [(7.66±2.17) vs (6.64±1.84) nmol/L], 2 h insulin [(129.81±145.49) vs (97.51±86.92) mU/L], total cholesterol [(5.35±0.89) vs (4.92±0.92) mmol/L], triglycerides [(1.55±1.28) vs (1.33±0.77) mmol/L], and low density lipoprotein cholesterol levels [(3.38±0.66) vs (3.14±0.71) mmol/L] were significantly higher in HA group, compared with NON-HA group (P<0.05). The initiated gonadotropin dose was higher in HA group than that in NON-HA group [(126.96±33.65) vs (137.60±38.12) U, P=0.001], but moderate-severe ovarian hyperstimulation syndrome (OHSS) rate was similar in two groups (P>0.05). The rates of implantation, clinical pregnancy, miscarriage, and live birth were comparable between the two groups (P>0.05). Also, in the subgroups, the rates of implantation, clinical pregnancy, live birth, and miscarriage were similar in HA group and NON-HA group. Conclusions: The risks of hormonal abnormality and glucose-lipid metabolic disorder were higher in PCOS women with HA, whereas satisfactory pregnancy outcomes could be achieved under proper ovarian stimulation undergoing IVF/ICSI-ET.

[Comparative study on imaging and clinical results of patellofemoral joint with kinematic alignment and mechanical alignment in total knee arthroplasty].

Objective: To investigate the changes of imaging parameters of patellofemoral joint after kinematic alignment total knee arthroplasty (KA-TKA) and mechanical alignment total knee arthroplasty (MA-TKA) and the effects on clinical outcomes. Methods: A retrospective analysis was performed on 227 patients diagnosed with knee osteoarthritis, 105 males and 122 females, with the age of (68.6±7.2) years (range: 52 to 86 years). The patients underwent unilateral TKA at the Department of Orthopaedic Surgery, Beijing Chaoyang Hospital, Capital Medical University, from January 2020 to July 2021. The patients were divided into the KA-TKA group (n=102) and the MA-TKA group (n=125) according to the alignment method. The Blackburne-Peel index, lateral patellofemoral angle, patellofemoral tilt angle and patellofemoral index of the two groups were recorded before and 6 months after operation. The frequency of lateral patellar retinaculum release, preoperative and postoperative knee range of motion (ROM), and Oxford knee score(OKS) were also recorded. The differences between preoperative and postoperative measurement data of each group were calculated. If the difference was in line with normal distribution, the independent sample t-test or t'-test was used for comparison between the two groups. If it did not conform to normal distribution, Mann-Whitney U test was used. Paired sample t-test was used for comparison before and after treatment. Chi-square test was used for comparation of categorical data between the two groups. Results: After 6 months of operation, the patellar tilt angles of the KA-TKA group and the MA-TKA group were (14.22±3.26)° and (13.35±2.27)°, and the lateral patellar angles were (9.73±4.86)° and (11.91±3.89)°, respectively. The change of lateral patellofemoral angle in the KA-TKA group was significantly less than that in the MA-TKA group ((1.68±4.86)° vs. (4.15±4.88)°, t=3.805, P<0.01). OKS and ROM were improved in the both groups at 6 months after operation (all P<0.05), but there were no statistic significance in preoperative and postoperative difference between the two groups (all P>0.05). The intraoperative lateral patellar retinacular release rate was 14.4% (18/125) in the MA-TKA group and 6.9% (7/102) in the KA-TKA group, with no statistical difference (χ2=3.256,P=0.071). Conclusions: There are greater patella lateral tilt at 6 months postoperatively in the KA-TKA group compared with the MA-TKA group, but this radiographic difference could not show differences of clinical outcomes and postoperative ROM, the frequency of intraoperative lateral patellar retinacular release wouldn't increase. Therefore, KA-TKA does not increase the difficulty of postoperative patellofemoral joint complications and intraoperative lateral patellar release.

[Clinical characteristics of 21 infertile women with non-classic 21-hydroxylase deficiency].

Objective: To investigate the clinical features of infertile women with non-classic 21-hydroxylase deficiency (21-OHD). Methods: The study enrolled 21 infertile women with non-classic 21-OHD in Sun Yat-sen Memorial Hospital of Sun Yat-sen University from January 2009 to December 2018. The clinical presentation, endocrine hormone, glucolipid metabolism and treatment outcome were retrospectively analyzed. The diagnosis of non-classic 21-OHD was comprehensively based on clinical and hormonal characteristics. Results: Among 21 cases, the age was (29.9±2.9) years, the mean age at menarche was (13.6±2.0) years, body mass index was (22.1±2.9) kg/m2, and 38% (8/21) had oligomenorrhea. Hirsutism was diagnosed in 3 cases (14%, 3/21). Clitoromegaly was seen in 14% (3/21) and polycystic ovarian morphology was found in 33% (7/21) of the patients. The mean serum level of basal progesterone was (11.3±21.0) nmol/L, with 48% (10/21) having high basal progesterone level; after therapy by glucocorticoid, the level of progesterone was (1.9±2.0) nmol/L. Serum 17-hydroxyprogesterone concentration was (66.4±123.6) nmol/L; after therapy by glucocorticoid, it was (2.4±1.8) nmol/L. In the study increased testosterone, androstenedione and dehydroepiandrosterone sulfate were present in 62% (13/21), 52% (11/21) and 43% (9/21), respectively; and 52% (11/21) of patients manifested androgen excess and basal progesterone elevation; androgen levels decreased after therapy by glucocorticoid. The pregnancy rate was 76% (16/21). Out of 19 pregnancies, 6/19 ended in spontaneous miscarriages. Conclusions: Infertile women with non-classic 21-OHD are characterized by hyperandrogenism and basal progesterone elevation, whereas gonad axis disorder is not apparent. After no response to conventional therapy, adult infertile women with non-classic 21-OHD could achieve a desirable pregnancy outcome with proper treatment of glucocorticoid.

Mitochondrial DNA Heteroplasmy of Hair Shaft Using HID Ion GeneStudioTM S5 Sequencing System.

ABSTRACT: Objective To study the heteroplasmy of the whole mitochondrial genome genotyping result of hair shaft samples using HID Ion GeneStudioTM S5 Sequencing System. Methods The buccal swabs and blood of 8 unrelated individuals, and hair shaft samples from different parts of the same individual were collected. Amplification of whole mitochondrial genome was performed using Precision ID mtDNA Whole Genome Panel. Analysis and detection of whole mitochondrial genome were carried out using the HID Ion GeneStudioTM S5 Sequencing System. Results The mitochondrial DNA sequences in temporal hair shaft samples from 2 individuals showed heteroplasmy, while whole mitochondrial genome genotyping results of buccal swabs, blood, and hair samples from the other 6 unrelated individuals were consistent. A total of 119 base variations were observed from the 8 unrelated individuals. The numbers of variable sites of the individuals were 29, 40, 38, 35, 13, 36, 40 and 35, respectively. Conclusion Sequence polymorphism can be fully understood using HID Ion GeneStudioTM S5 Sequencing system.

[Clinical analysis of infective endocarditis with septic pulmonary embolism].

Objective: To analyze the clinical features of infective endocarditis with septic pulmonary embolism, and to improve the diagnosis and treatment of the disease. Methods: Retrospective analysis of clinical manifestations, blood culture, echocardiography, chest CT and diagnosis and treatment of 16 patients with infective endocarditis and septic pulmonary embolism in the First Affiliated Hospital of Zhengzhou University from January 2010 to June 2017 was carried out, and the clinical features of the disease were summarized. Results: Out of the 16 patients, 4 were males and 12 were females, aged (29.5±9.7) years old, 12 patients were with congenital heart disease. The main clinical manifestations were fever, cough, dyspnea, hemoptysis. There were 7 cases with positive blood culture, and 6 of them were Streptococcus. Echocardiography revealed cardiac vegetations, and all were from right heart. Chest CT was characterized by multiple pulmonary lesions, mainly characterized by multiple nodules and small patch shadows. All 16 patients were initially misdiagnosed as pneumonia. All patients were treated with antibiotics intravenously, 13 patients underwent cardiac surgery, 6 patients were cured, 8 patients improved, and 2 patients were discharged automatically. Conclusions: Infective endocarditis with septic pulmonary embolism is easily misdiagnosed. In patients with fever and multiple shadows in the lungs, the possibility of right heart infective endocarditis should be considered. The diagnosis can be confirmed once cardiac vegetations are found by echocardiography, and repeated echocardiographic examinations can be performed if necessary. The prognosis of patients treated with surgery as early as possible on the basis of anti-infection is good.

[Clinical significance of anti-interferon -α2 and anti-interferon -ω antibody in myasthenia gravis].

Objective: To measure the levels of anti-interferon (IFN) -α2 antibody (IFN-α2-Ab) and anti-IFN-ω antibody(IFN-ω-Ab) and investigate their potential roles in patients with myasthenia gravis (MG). Methods: Radioimmunoprecipitationassay (RIPA) based on (125)I-labelled IFN-α2, -ω, acetylcholine receptor (AChR) and muscle-specific kinase (MuSK) were used to detect anti-IFN-α2-Ab, anti-IFN-ω-Ab, anti-AChR-Ab and anti-MuSK-Ab respectively in MG patients from Tianjin Medical University General Hospital, during the year of 2013. Relationship between IFN-α2-Ab and clinical characteristics in patients with MG were analyzed. Results: With the RIPA, anti-IFN-α2-Ab was positive in 11 (22%) patients with MG, among whom, 8 were thymoma associated MG (TAMG) and 3 were late onset MG (LOMG). The double positive one was among the LOMG who was a 76-year-old ocular MG patient. Serum anti-IFN-α2-Ab levels in different subgroups of MG had no significant difference (P>0.05). MG patients with anti-IFN-α2-Ab complicated with less other autoimmune disease. Serum anti-IFN-α2-Ab levels negatively correlated with QMG score in TAMG. Conclusions: MG patients with anti-IFN-α2-Ab complicate with less other autoimmune disease. The higher serum IFN-α2-Ab levels in TAMG, the slighter symptoms present.

[Clinical characteristics of lung NK/T cell lymphoma misdiagnosed as pneumonia].

Objective: To investigate the clinical features of lung natural killer (NK)/T cell lymphoma. Methods: The clinical data of patients with lung NK/T-cell lymphoma confirmed by pathology who were hospitalized due to lung shadow and initially treated as pneumonia from the First Affiliated Hospital of Zhengzhou University was collected from June 2013 to January 2019. The clinical manifestations, laboratory tests, chest CT findings, treatment procedures, outcomes, and misdiagnosis were retrospectively analyzed. Results: Among the 15 enrolled patients with lung NK/T-cell lymphoma, 5 were primary and the other 10 were secondary. There were 8 males and 7 females, aged 20-76 years. Fever presented in 14 cases, dyspnea was observed in 4 cases and 2 cases had cough. A total of 12 cases had leukopenia, 10 cases had mononuclear cell increase, 10 cases had liver dysfunction, 13 cases had elevated serum lactate dehydrogenase (LDH), and 2 cases of primary lung NK/T-cell lymphoma had increased pleural LDH and adenosine deaminase. Five cases of primary lung NK/T-cell lymphoma had multiple lung lesions, with diffuse multiple solid shadows, ground glass and patchy shadows, some with nodules; 10 cases of secondary lung NK/T-cell lymphoma showed nodular masses, some were accompanied by patchy shadows, and most patients had multiple lesions scattering in the lung. All the fifteen patients were misdiagnosed as pneumonia in the early stage, and then were diagnosed by percutaneous lung biopsy or transbronchial biopsy. Six patients underwent chemotherapy, of which 1 achieved complete response, 4 were in follow-up and 1 died of respiratory failure; 5 patients died of respiratory failure after diagnosis and 4 patients discharged and died within half a year. Conclusions: The lung NK/T-cell lymphoma are rare. The clinical manifestations and pulmonary imaging findings are indistinguishable from pulmonary inflammatory lesions. They are easily misdiagnosed as pneumonia. Early CT-guided percutaneous lung biopsy or transbronchial biopsy can improve the diagnostic accuracy.

[Application of RapidHIT™ 200 System in Forensic Medicine].

OBJECTIVES: To validate the analysis capability of RapidHIT™ 200 system for four kinds of routine forensic samples and the recyclable capability of template, template DNA and PCR products in the process of twice duplicate detection. METHODS: The buccal swabs underwent the test twice by RapidHIT™ 200 system, and the template DNA and PCR products that arose in the system were also tested for two times. After four kinds of routine forensic samples were detected by RapidHIT™ 200 system, the follow-up tests of the template, template DNA and PCR products that arose in the system were performed. RESULTS: The STR loci could be detected in the buccal swabs by the system for the first time. However, part of the STR loci lost during the second test. And the peak value obtained in the second test was significantly reduced than the one in the first time. The average STR loci detection rates of the template DNA and PCR products were both less than 50% in the second test, which were significantly reduced than that in the first test. In addition, the analysis capability of the system for the tissues and buccal swabs was better than that for the blood and cigarette butts. Compared with the first test, the STR loci detection rate of the tested items, template DNA and PCR products decreased with the numbers of tests. CONCLUSIONS: RapidHIT™ 200 system is more effective in retesting buccal swabs than other samples, whereas the items, DNA template, PCR products obtained in the first and second time cannot be directly used for the further application and study of forensic medicine.

Dietary luteolin activates browning and thermogenesis in mice through an AMPK/PGC1α pathway-mediated mechanism.

BACKGROUND: Two brown-like adipocytes, including classical brown adipocytes from brown adipose tissues and beige cells from white adipose tissues, regulate thermogenesis. The developmental and functional induction of brown-like cells provides a defense against obesity and associated metabolic diseases. Our previous study suggests dietary luteolin can improve diet-induced obesity and insulin resistance in mice. Here we further elucidated the action of the natural flavonoid on energy expenditure and adaptive thermogenesis. METHODS: Five-week-old male C57BL/6 mice were fed low-fat diet (LFD), high-fat diet (HFD) and HFD supplemented with 0.01% luteolin. After 12 weeks, their energy expenditure were detected using a combined indirect calorimetry system. Moreover, thermogenic program and associated molecular regulators were assessed in adipose tissues. In another independent study, even-aged mice were fed LFD and luteolin-containing LFD for 12 weeks, and their energy expenditure and thermogenic program were also investigated. Finally, differentiated primary brown and subcutaneous adipocytes were used to identify the critical participation of AMPK/PGC1α signaling in luteolin-regulated browning and thermogenesis. RESULTS: In mice fed either HFD or LFD, dietary luteolin supplement increased oxygen consumption, carbon dioxide production and respiratory exchange ratio. The enhancement in energy expenditure was accompanied by the upregulation of thermogenic genes in brown and subcutaneous adipose tissues. Meanwhile, several important AMPK/PGC1α signaling molecules were activated by dietary luteolin in the tissues. Further, luteolin treatment directly elevated thermogenic gene expressions and activated AMPK/PGC1α signaling in differentiated primary brown and subcutaneous adipocytes, whereas AMPK inhibitor Compound C reversed the efficiencies. CONCLUSIONS: Dietary luteolin activated browning and thermogenesis through an AMPK/PGC1α pathway-mediated mechanism.

Association between interleukin-21 gene polymorphisms (rs12508721) and HBV-related hepatocellular carcinoma.

Interleukin-21 (IL-21), as a multifunctional cytokine, plays an important role in many diseases, such as cancer, inflammatory and autoimmune diseases. We aimed to investigate the relationship between polymorphisms of IL-21 gene and susceptibility of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) in a Chinese population. Studied subjects were divided into three groups: 100 patients with HBV-related HCC, 115 patients with chronic HBV infection and 127 healthy controls. Genomic DNA was isolated from peripheral blood, and the polymerase chain reaction-ligase detection reaction (PCR-LDR) method was used to genotype the SNPs (rs2221903, rs907715 and rs12508721) within IL-21 gene. Our results showed that IL-21 polymorphisms were associated with the risk of HCC and chronic HBV infection when compared with healthy controls. The rs2221903A/G AG genotype was associated with a higher risk of chronic HBV infection when compared with healthy controls [AG versus AA + GG, P = 0.036, OR = 1.898, 95%CI = 1.038-3.471]. The rs12508721C/T TT genotype was related with a lower risk of chronic HBV infection and HBV-related HCC than in healthy controls [TT versus CT + CC, P = 0.026, OR = 0.451, 95%CI = 0.221-0.920; P = 0.049, OR = 0.482, 95%CI = 0.231-1.005]. No significant difference in the genotype and allele distrubutions of rs907715G/A SNP was observed in the HBV-related HCC group, chronic HBV-infected group and the healthy control group when compared to each other. Our findings suggest that the rs12508721T/C and rs2221903A/G polymorphisms of IL-21 gene are associated with the susceptibility of HBV-related HCC and chronic HBV infection. The genetic variant may in fact cause protection against the HBV-related HCC. However, the function in these SNPs of IL-21 gene needs to clarify the mechanisms involved in the pathogenesis of HBV-related HCC further.

[Predictive value of deubiquitination enzymes USP37 in the prognosis of breast cancer].

OBJECTIVE: To explore the expression of Ubiquitin-specific processing enzyme 37(USP 37)in breast cancer and its association with the prognosis of breast cancer. METHODS: In this study, the method of Western blot was utilized to examine the expression of USP37 protein in breast cancer fresh tissue. Immunohistochemistry (IHC) was used to determine the expression of USP37 in 533 cases of breast cancer. Receive operating characteristic (ROC) curve analysis was employed to determine a cut-off score for USP37 expression. For validation, the ROC-derived cut-off score was subjected to analysis the association of USP37 expression with cancer patient outcome and clinical characteristics. RESULTS: USP37 protein was mainly located in cytoplasm of the cell, occasionally in nucleus by immunohistochemistry. In the normal breast tissue, USP37 was not expressed or with low expression level, while in the 533 cases of breast cancer, the high USP37 expression was detected in 50.7% samples (270/533) with corresponding low or negative expression rate 49.3% (263/533). We found that USP37 was highly expressed in primary breast cancer tissues compared to paired adjacent non-cancerous tissues. High expression of USP37 was associated with breast cancer node classification, molecular classification and proliferation biomarker Ki67. Both univariate and multivariate analyses further revealed that USP37 expression was an independent poor prognostic parameter for overall survival (OS), recurrence-free survival (RFS) and metastasis-free survival (MFS) in breast cancer. Furthermore, USP37 expressions divided the luminal and triple negative breast cancer into different prognosis subgroups. CONCLUSION: Our findings first provide evidences that high expression of USP37 served as an independent molecular biomarker for poor prognosis in breast cancer.

[Clinical analysis of fourteen cases with neuromyelitis optica spectrum disorders].

Objective: To analysis the clinical manifestation, radiological data, laboratory findings of neuromyelitis optica spectrum disorders (NMOSD) to help physicians have a deeper understanding of the disease. Methods: The clinical manifestations, radiological data, laboratory findings of NMOSD patients from People's Hospital of Zhengzhou University between June 2015 and June 2016 were retrospectively analyzed. Results: Fourteen patients with NMOSD were enrolled, consisting of 3 males and 11 females. The onset age ranged from 15 to 62 years, in average 39.64. Among them, there was 1 case with history of hyperthyroidism, 3 cases with Sjögren syndrome (SS). Major clinical manifestations were vision loss in 4 cases, nausea and vomiting in 4 cases, limbs numbness and weakness in 6 cases. All the 14 patients were received the cerebrospinal fluidcell surface antigen antibody detection, and the results were strong positive 6 cases (6/14), positive 7 cases (7/14), weak positive 1 case (1/14). Abnormal signals in the thirdventricle, midbrain aqueduct, oblongata, pons and cervical, thoracic spinal cord were found by Magnetic Resonance Imaging (MRI). Fourteen patients were treated with glucocorticoid, among them 8 received immunosuppressive therapy. All the 14 patients were followed up, and 9 patients had different degrees of sequelae, including vision decline in 4 cases, numbness and weakness in 5 cases. Conclusions: Neuromyelitis optica spectrum disorders frequently occur in young female. Clinicians need to consider the possibility of NMOSD for acute or subacute blurred vision, nausea, vomiting, numbness and weakness of limbs or trunk and psychiatric symptoms. NMOSD can not be easily excluded for those special patients with atypical clinical manifestations, and relevant inspections should be performed as far as possible in order to make the correct diagnosis.

[Association between interleukin-18 gene polymorphisms and hepatocellular carcinoma caused by hepatitis B virus].

OBJECTIVE: To investigate the association between the single nucleotide polymorphisms (SNPs) IL-18-137G/C (rs187238) and IL-18-607A/C (rs1946518) in interleukin-18 (IL-18) gene and hepatocellular carcinoma (HCC) caused by the hepatitis B virus (HBV). METHODS: The subjects were divided into HBV-related HCC group (109 patients), chronic HBV infection group (113 patients), and healthy control group (127 patients). The polymerase chain reaction-ligase detection reaction (PCR-LDR) was used to determine the alleles and genotypes of the two SNPs IL-18-137G/C and IL-18-607A/C. The t-test and chi-square test were used for baseline data. The chi-square test was used to investigate the differences in genotype and allele frequencies across the three groups. Non-conditional logistic regression analysis was used to compare the odds ratios (ORs) and 95% confidence intervals (CIs) for different genotypes/alleles in predicting the risk of HBV-related HCC. RESULTS: The HBV-related HCC group showed significantly higher AA genotype and A allele frequencies of the SNP IL-18-607A/C than the healthy control group (AA genotype frequency: 29.4% vs 18.1%, χ (2) = 4.152, P < 0.05; A allele frequency: 54.6% vs 44.1%, 5.169, P < 0.05), which were positively correlated with the risk of HBV-related HCC (AA genotype frequency: OR = 1.879, 95% CI: 1.020-3.464; A allele frequency: OR = 1.524, 95% CI: 1.059-2.193). The chronic HBV infection group had a significantly higher A allele frequency of the SNP IL-18-607A/C than the healthy control group (54.0% vs 44.1%, χ (2) = 4.680, P < 0.05), which was positively correlated with the risk of chronic HBV infection (OR = 1.487, 95% CI: 1.037-2.132). The genotype and allele frequencies of the SNP IL-18-607A/C showed no significant differences between the HBV-related HCC group and the chronic HBV infection group (P > 0.05). The genotype and allele frequencies of the SNP IL-18-137G/C showed no significant differences between any two groups of the three groups (P > 0.05). CONCLUSION: The AA genotype and A allele frequencies of the SNP IL-18-607A/C are positively correlated with the morbidity of HBV-related HCC, and the A allele frequency of the SNP IL-18-607A/C is positively correlated with the morbidity of chronic HBV infection.

Cloning and characterization of a benzoic acid/salicylic acid carboxyl methyltransferase gene involved in floral scent production from lily (Lilium 'Yelloween').

In lily flowers, the volatile ester methyl benzoate is one of the major and abundant floral scent compounds; however, knowledge regarding the biosynthesis of methyl benzoate remains unknown for Lilium. In this study, we isolated a benzoic acid/salicylic acid carboxyl methyltransferase (BSMT) gene, LiBSMT, from petals of Lilium 'Yelloween'. The gene has an open reading frame of 1083 base pairs (bp) and encodes a protein of 41.05 kDa. Sequence alignment and phylogenetic analyses of LiBSMT revealed 40-50% similarity with other known benzenoid carboxyl methyltransferases in other plant species, and revealed homology to BSMT of Oryza sativa. Heterologous expression of this gene in Escherichia coli yielded an enzyme responsible for catalyzing benzoic acid and salicylic acid to methyl benzoate and methyl salicylate, respectively. Quantitative real-time polymerase chain reaction analysis showed that LiBSMT was preferentially expressed in petals. Moreover, the expression of LiBSMT in petals was developmentally regulated. These expression patterns correlate well with the emission of methyl benzoate. Our results indicate that LiBSMT plays an important role in floral scent methyl benzoate production and emission in lily flowers.

Development of expressed sequence tag-simple sequence repeat markers for Chrysanthemum morifolium and closely related species.

With the development of chrysanthemum breeding in recent years, an increasing number of wild species in genera related to Chrysanthemum were introduced to extend the genetic resources and facilitate the genetic improvement of chrysanthemums via hybridization. However, few simple sequence repeat (SSR) markers are available for marker-assisted breeding and population genetic studies of chrysanthemum and closely related species. Expressed sequence tags (ESTs) in public databases and cross-species transferable markers are considered to be a cost-effective means for developing sequence-based markers. In this study, 25 EST-SSRs were successfully developed from Chrysanthemum EST sequences for Chrysanthemum morifolium and closely related species. In total, 4164 unigene sequences were assembled from 7180 ESTs of chrysanthemum in GenBank, which were subsequently used to screen for the presence of microsatellites with the SSRIT software. The screening criteria were 8, 5, 4, and 3 repeating units for di-, tri-, tetra-, and penta- and higher-order nucleotides, respectively. Moreover, 310 SSR loci from 296 sequences were identified, and 198 primer pairs for SSR amplification were designed with the Primer Premier 5.0 software, of which 25 SSR loci showed polymorphic amplification in 52 species and varieties belonging to Chrysanthemum, Ajania, and Opisthopappus. The application of EST-SSR markers to the identification of intergeneric hybrids between Chrysanthemum and Ajania was demonstrated. Therefore, EST-SSRs can be developed for species that lack gene sequences or ESTs by utilizing ESTs of closely related species.

Association between PAI-1 polymorphisms and plasma PAI-1 level with sepsis in severely burned patients.

We investigated the association between plasminogen activator inhibitor-1 (PAI-1) polymorphisms and plasma PAI-1 level with sepsis in severely burned patients. A total of 182 patients with burn areas lager than 30% of the body surface area were enrolled in this study. Peripheral blood samples were obtained from 103 patients with sepsis (sepsis group) and 79 patients without sepsis (control group). An allele-specific polymerase chain reaction assay was used to determine PAI-1 polymorphism 4G/5G distribution. Plasma PAI-1 levels were detected using an enzyme-linked immunosorbent assay. The frequency of the 4G/4G genotype and the 4G allele frequency in the sepsis group were 42.7 and 62.1% respectively, which were significantly higher than those in the control group (P < 0.05). Sepsis patients had a significantly higher plasma PAI-1 level than the control group (P < 0.05). Compared with the 5G/5G genotype, PAI-1 concentrations were significantly higher in the 4G/4G genotype (P < 0.05). The study indicates that the 4G/5G promoter polymorphism of PAI-1 gene may be related to the susceptibility to burn sepsis and that the 4G/4G genotype may be an important genetic risk factor of burn sepsis. Additionally, PAI-1 concentrations in the serum are increased in patients with burn sepsis.

Safety of recombinant human granulocyte-macrophage colony-stimulating factor in healing pediatric severe burns.

We explored the safety of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) for healing burns in children. Subjects were randomly assigned to two groups: the experimental group received external rhGM-CSF gel, and the control group received rhGM-CSF gel matrix components, applied to the burn surface. Neither group was given any other drugs that promote wound healing. Each day we recorded the pulse, body temperature, and respiration status in the two groups. We detected the blood routine, urine routine, and hepatic and renal function before the patients received drug treatment and after 72 h. The wound scab and healing states in the two groups were recorded every 4 days to evaluate wound healing rate and time taken for complete healing. Adverse reactions and their rate of occurrence were also recorded. The median time of healing was 15 days in the experimental group and 19 days in the control group (log-rank χ(2) = 5.139, P < 0.05). After 10 days, the experimental group healing rate was consistently higher than that of the control group (significantly different using intuitive analysis), suggesting the experimental group method was more effective. There were no obvious adverse reactions. There was no significant difference between the blood routine, urine routine, and liver and kidney function in the two groups before the treatment and after 3 days (P > 0.05). Compared with saline treatment of severe burns, rhGM-CSF can effectively shorten the healing time without significant adverse reactions, and is an effective and safe treatment for burns in children.

Screening of molecular markers linked to dwarf trait in crape myrtle by bulked segregant analysis.

Plant height is one of the most important traits of plant architecture as it modulates both economic and ornamental values. Crape myrtle (Lagerstroemia indica L.) is a popular ornamental woody plant because of its long-lasting mid-summer bloom, rich colors, and diversified plant architecture. These traits also make it an ideal model of woody species for genetic analysis of many ornamental traits. To understand the inheritance of plant height and screen for genes modulating plant height in Lagerstroemia, segregation of the plant height trait was analyzed using the F1 population of L. fauriei (standard) x L. indica 'Pocomoke' (dwarf) with 96 seedlings, while dwarf genes were screened using the bulked segregant analysis method, combined with 28 amplified fragment length polymorphism primers and 41 simple sequence repeat primers. The results showed that the dwarf trait of crape myrtle was controlled by a major gene and modified by minor genes. An amplified fragment length polymorphism marker, M53E39-92, which was 23.33 cM from the loci controlling the dwarf trait, was screened. These results provide basic information for marker-assisted selection in Lagerstromia and cloning of dwarf genes in future studies.

Study of Hgp44 from Porphyromonas gingivalis on inducing HUVECs to secrete IL-6 and IL-8.

The aim of this study was to clone, express the gene of Hgp44 in adhesin domains of gingipains from Porphyromonas gingivalis and purify the protein. Furthermore, the effect of Hgp44 from P. gingivalis on inducing HUVECs to secrete IL-6 and IL-8 was evaluated. The Hgp44 gene fragment was amplified by polymerase chain reaction, and then inserted into the cloning vector pMD18-T and linked with a prokaryotic expression vector pET22b to construct the recombinant expression plasmid pET22b-Hgp44. Fusion protein expression was induced by IPTG, and it was purified by immobilized metal-chelating affinity chromatography (IMAC) using an Ni(2+) matrix column. HUVECs were cultured in vitro and different concentrations of Hgp44 were added to confluent HUVEC monolayers and incubated for 2, 8 and 24 h. We extracted the supernatants and then used ELISA kits to test the changes in IL-6 and IL-8 levels. Finally, a 1100-bp fragment was successfully amplified, and the expression of the fusion protein was examined by SDS-PAGE and Western blot analysis, and the data showed that the protein was 44 kDa in size and expressed mostly in the form of inclusion bodies. The purification of the fusion protein was achieved using Ni(2+) affinity chromatography. About 3.5 mg/L fusion protein was obtained. Hgp44 could induce HUVECs to secrete IL-6 and IL-8 levels, which were remarkably increased. In a word, Hgp44 was successfully expressed in a prokaryotic expression system and purified by IMAC using the Ni(2+) matrix column. The effect of Hgp44 in inducing HUVECs to secrete IL-6 and IL-8 was demonstrated.