Volume 94  ·   Issue 14  ·  2014

Prostate cancer is the malignant tumor with the highest incidence in men in Europe and America, and ranks second in male cancer mortality in the United States. The incidence of prostate cancer is positively correlated with age^[1]。 With the extension of population life expectancy, diet structure and lifestyle changes, the incidence of prostate cancer in China is constantly increasing. Currently, the only marker clinically used for screening of prostate cancer and monitoring of relapse after treatment is prostate specific antigen (PSA). Although PSA-based screening allows more early patients to be diagnosed and treated, 23% to 42% of patients found by screening are occult prostate cancer with slow progression, low aggression, and no clinical symptoms. Two prospective studies of whether PSA reduces the risk of death from prostate cancer in the United States and Europe both acknowledge that PSA screening leads to overtreatment in some cases, adding unnecessary pain, injury and financial burden to patients^[2,3]。 Although screening also reduces disease staging, it does not substantially improve the overall fatality rate. Many occult subclinical tumors are diagnosed and treated, shifting the focus of research from previous risk gene susceptibility studies to early diagnosis of aggressive prostate cancer and molecular signature studies of fatal prostate cancer. For patients with biochemical recurrence suggested by elevated PSA after surgery or radiotherapy, there are limited tools to understand the risk of disease progression and guide the course of treatment. The study of high-risk markers of recurrence is of great significance to understand the mechanism of occurrence and exacerbation of prostate cancer, which is conducive to clinicians to better grasp the surgical indications. Positive measures such as strengthening follow-up, adjuvant endocrine therapy or radiotherapy for patients with high-risk of recurrence can benefit patients. Many studies have attempted to identify high-risk markers of prostate cancer progression, but most of them have not achieved the intended purpose, and no markers that can be routinely used in clinical practice have been found. Some views on the research of markers of recurrence and progression of prostate cancer are discussed.

To explore the magnetic resonance imaging (MRI) manifestations of early prostatic carcinoma (PCa) in central zone(CZ).

A total of 284 cases of PCa were scanned by MRI while 24 cases of carcinoma in 28 PCa focus were in central zone. Their T1WI, T2WI, DWI and ADC value characteristics were analyzed respectively and the completeness of annular low signal between central and peripheral zones were observed. At the same time, 45 cases of PCa were located in peripheral zone(PZ). Twenty cases of benign prostatic hyperplasia (BPH) were comparatively analyzed. All cases received a histological diagnosis.

Most cases of early PCa in CZ demonstrated ill-defined margin, amorphous shape and equal signal intensity on T1WI, low signal intensity on T2WI and high signal intensity on DWI. The mean ADC value for PCa focus was (0.867±0.074)×10^-3 mm²/s and PCa foci were enhanced slightly. In these cases, most annular low signals between CZ and PZ were destroyed. The difference of ADC values between PCa in CZ and PZ groups((0.914±0.066) ×10^-3 mm²/s)had no statistic significance(P>0.05), but the ADC values between PCa and BPH in CZ groups((1.517±0.315) ×10^-3 mm²/s)had statistically significant difference(P<0.05).

Some reliable diagnostic characteristics on MRI may be used for an early diagnosis of PCa in CZ.

To explore the clinical significance of detecting serum mRNA of telomerase in the diagnosis of prostate cancer.

Serum samples were collected from 29 patients with prostate cancer and 29 age-matched patients with cardiovascular or metabolic disease as non-tumor controls. Sera samples from 15 healthy age-matched subjects were used as healthy control. Detection of serum telomerase mRNA was performed with real-time reverse transcriptase quantitative polymerase chain reaction (PCR).

Serum telomerase mRNA was detectable in 89.7%(26/29) patients with prostate cancer, but almost undetectable in non-tumor (6.9%, 2/29) and healthy control groups(1/14).

Using real-time PCR for detecting serum telomerase mRNA may be an auxiliary method for diagnosing and monitoring of prostate cancer.

To retrospectively analyze the effects of bladder neck preservation (BNP) and reconstruction of rhabdosphincter (RS) on urinary continence recovery for patients after laparoscopic radical prostatectomy (LRP).

Bladder neck preservation plus reconstruction of rhabdosphincter(BNP+ RS)was employed in 18 prostate cancer patients after LRP from January 2011 to December 2012. Eighteen contemporary cases of routine LRP were selected for control group. The operative duration, blood loss volume, postoperative surgical margins, urine leakage and continence rate were compared between two groups.

LRP was successfully performed through extraperitoneal space in all 36 cases. There was no conversion into open surgery. The median operative duration, blood loss volume, positive surgical margins and urine leakage rate in BNP+ RS and control groups were 164(125-245)versus 142(95-210)min, 195(80-550)versus 230(120-400)ml, 3 versus 2 cases and 0 versus 2 cases respectively. At 3, 6, 12 months, the continence rates in BNP+ RS and control groups were 12 cases (66.7%) versus 5 cases(13.9%, P=0.044), 15 cases(83.3%) versus 11 cases(30.6%, P=0.264)and 16 cases (88.9%) versus 13 cases(36.1%, P=0.402)respectively. A significant higher continence rate in the first 3 months postoperatively was found in BNP+ RS group than control group.

BNP plus RS reconstruction after LRP can significantly improve continence in the first 3 months after operation.

To screen the plasma microRNA(miRNA)profile of immune thrombocytopenia (ITP) patients.

Agilent 19.0 miRNA microarray was used to detect the expression profile of miRNA in plasma from 25 ITP patients and 20 healthy controls from June 2012 to September 2013. The software programs of TargetScan and miRanda were used for predicting target genes associated with differential miRNA. Then gene ontology (GO) and pathway analysis were performed to explore the genes and pathways involved in the pathogenesis of ITP. And differential miRNA was validated by real-time quantitative polymerase chain reaction (PCR).

A genome-wide miRNA array revealed 29 differential miRNAs in the plasma samples of ITP patients including 15 up-regulated and 14 down-regulated miRNA. A total of 608 potential genes were predicted by TargetScan and miRanda. GO result showed that there were 475(78.12%), 491(80.76%) and 533(87.66%) genes respectively involved in biological process, molecular function and cellular component. Enrichment test showed 9 GO terms had significant difference (P<0.05). Pathway analysis showed that 157 pathways were associated with 608 genes. Enrichment test showed 25 pathways had significant difference (P<0.05). As revealed by real-time PCR, the expressions of miRNA4778-5p and miRNA4800-5p became obviously up-regulated while those of miRNA4707-5p, miRNA4721, miRNA3620-3p and miRNA378i decreased (allP<0.05). The results agreed with those of microarray.

The plasma differential miRNA profiles are identified in ITP patients. And miRNA is involved in calcium signaling pathway and T cell receptor signaling pathway may be associated with ITP pathogenesis.

To evaluate the imaging efficacies after Salter innominate osteotomy for developmental dysplasia of the hip (DDH).

A total of 51 DDH patients with 70 hips were recruited between March 2011 to August 2012. All of them underwent Salter innominate osteotomy. Preoperative radiographs including acetabular index (AI), sharp angle (SA), obturator angle and obturator height were measured. And postoperative radiographs were taken at 2 days, 3 months, 6 months and 1 year to measure the parameters of AI, sharp acetabular angle, obturator angle, obturator height, shifting index and descending angle of distal iliac end. Paired t test was used for statistical analyses.

The average correction of AI was 35.5°±2.3° preoperatively versus 14.6°±1.6° postoperatively (P=0.000). The average correction of Sharp angle was 50.6°±3.7° preoperatively versus 33.1°±3.2° postoperatively (P=0.000). The average correction of obturator angle was 58.3°±3.9° preoperatively versus 39.5°±2.8° postoperatively(P=0.000). The average correction of obturator height was(1.65±0.26)cm preoperatively versus(1.10±0.14)cm postoperatively (P=0.000). At 3 months, 6 months and 1 year post-operation, the average values of AI, SA, obturator angle and obturator height continued to fall. Both of them showed statistical improvement. Shifting index and descending angle of distal iliac end were 30.1%±3.6% and 31.3°±4.1° on average. Six (8.57%) Shenton lines were bad.

The imaging evaluations of acetabulum and pelvis show marked improvements until 1 year in DDH patients after Salter innominate osteotomy. It provides data reference for operative evaluations through measuring the imaging parameters of truncation bone.

To assess the value of cardiopulmonary exercise test in the evaluation of cardiac function in patients with chronic left heart failure caused by dilated cardiomyopathy.

Fifty-three inpatients aged 18 year and over with chronic left heart failure caused by dilated cardiomyopathy at Fuwai Hospital from October 2010 to October 2011 were selected and divided into 2 groups according to the New York Heart Association (NYHA) heart function classification. One group had 20 cases for class Ⅱ and another 33 cases for class Ⅲ-Ⅳ. All of them received cardiopulmonary exercise tests. Synchronous measurement and record of gas exchange indices were taken during every breath, and so were heart rate, blood pressure, electrocardiogram and blood oxygen saturation. At the same time, other routine tests were also performed. After exercise test, anaerobic threshold and peak oxygen consumption indices were calculated and statistically analyzed. They also received subsequent follow-ups of 1 day, 1 week, 1 month, 6 months and 1 year, including activities, clinical manifestations and cardiac adverse events.

At baseline, the differences in gender, age, body mass index, concurrent diseases, left ventricular end-diastolic diameter, left ventricular ejection fraction and serum creatinine had no statistical significance (P>0.05). Compared with cardiac function class Ⅱ group, the class Ⅲ-Ⅳ group had higher left atrial diameter, level of amino terminal pro-B-type natriuretic peptide (NT-proBNP), incidence of atrial fibrillation ((51.4±7.5) vs (43.6±7.7) mm, (2 607±1 782) vs (1 312±901) μg/L, 42.4%(14/33) vs 5.0%(1/20)) and lower glomerular filtration rate, peak oxygen consumption, levels of anaerobic threshold ((72±20) vs (97±23) ml/min, (13.7±2.6) vs (20.5±3.6) ml·min^-1·kg^-1, (10.7±1.5) vs (13.3±2.1) ml·min^-1·kg^-1) (all P<0.01). And NT-proBNP and cardiac function classification showed a positive correlation (OR=1.002, P=0.003) while peak oxygen consumption, anaerobic threshold and cardiac function classification were negatively correlated (OR=0.736, 0.608; P=0.011, 0.001).

Cardiopulmonary exercise test objectively reflects the cardiopulmonary reserve of heart failure patients with dilated cardiomyopathy. And the parameters of anaerobic threshold and peak oxygen consumption may reflect the patient's motor ability quantitatively and accurately.

To explore the methylation status of shp1 gene in plasma DNA from patients with diffuse large B cell lymphoma (DLBCL) and discuss its possible application in molecular diagnosis and targeted therapy of the disease.

Methylation-specific polymerase chain reaction (MSP) was used to detect the methylation status of shp1 gene in plasma and peripheral blood leukocytes (PBLs) of 35 DLBCL patients. The formaldehyde-fixed, paraffin-embedded (FFPE) tumor tissue samples were collected from 28 DLBCL patients, 6 patients of benign lymphoid hyperplasia and 13 healthy volunteers were selected as nonmalignant controls from January 2012 to December 2013. Methylation frequencies of shp1 gene in different groups were compared and the associations of shp1 methylation status with clinicopathological characteristics were analyzed.

No methylation of shp1 was detected in any of the 19 nonmalignant controls. The methylation rate of shp1 in plasma, PBLs and FFPE tumor tissues from patients with DLBCL was 51.4%(18/35), 28.6%(10/35) and 64.3%(18/28) respectively; there was a high methylation consistency of shp1 between plasma and FFPE tumor tissues (κ=0.78, P=0.00). However, methylation consistency was lower between PBLs and FFPE tumor tissues (κ=0.36, P=0.01). Methylation of shp1 was frequently detected in plasma and FFPE tumor tissues samples from patients with a high serum level of lactate dehydrogenase(13/16 vs 5/19, 11/12 vs 7/16, P=0.02, 0.04). However, no such association was detected in PBLs(P=0.14).

Methylation of shp1 in plasma DNA can represent shp1 methylation status in tumor tissue. And it may serve as a promising biomarker in aiding DLBCL diagnosis and guiding targeted therapy.

To explore the correlation between the expression of neuropilin-2 (NRP2) and tumor-associated lymphangiogenesis and clinicopathological parameters in gastric carcinoma (GC).

All GC, margin and normal tissues of 65 cases through surgical resection were retrospectively collected at our hospital and Tangshan People's Hospital from October 2011 to October 2013. The expressions of NRP2 in GC, tumor margin and normal gastric tissues were analyzed by immunohistochemistry. Lymphangiogenesis was marked with D2-40 monoclonal antibody and microlymphatic density (MLD) counted. And the morphology of lymphatic vessels was observed.

A positive expression of NRP2 was found in GC (n=40, 61.5%) and it was significantly higher than that in tumor margin (n=32, 49.2%) and normal tissues (n=3, 4.6%) (P<0.05). But the counts of MLD in both GC (12±5)/mm² and normal region (23±6)/mm² were significantly lower than that of tumor margin region (44±13)/mm² (P<0.05). In addition, the morphologies of lymphatic vessels were different among 3 groups. Also NRP2 expression and MLD count were correlated with differentiation degree, infiltrative degree, lymphatic vessel invasion, lymph node metastasis and TNM staging of GC (allP<0.05). The expression intensity of NRP2 in tumor margin tissues was positively correlated with MLD (r=0.517, P=0.000).

The expression of NRP2 in GC may modulate lymphangiogenesis and lymphatic metastasis. And it may play a key role in tumorigenesis, progression and prognosis predicating in GC.

To explore the associations of ulcerative colitis (UC) with vitamin D receptor (VDR) gene polymorphisms and serum levels of 25-hydroxyl vitamin D[25(OH)D].

From July 2004 to July 2013, a total of 404 UC patients were recruited from 4 hospitals of Wenzhou City. A total of 612 controls were collected from Health Examination Center of Second Affiliated Hospital of Wenzhou Medical University. Four single nucleotide polymorphisms of VDR (Fok Ⅰ, Bsm Ⅰ, Apa Ⅰ, Taq Ⅰ) were detected by mini-sequencing technique. The frequencies of minor allele and genotype of VDR were compared between UC patients and the controls by χ ² test and Bonferroni correction. Moreover, 75 UC patients and 120 gender-and age-matched healthy controls during the corresponding period were randomly selected for determining the serum levels of 25(OH)D by electrochemiluminescence immunoassay and were compared by Student's test.

After Bonferroni correction, mutant allele and genotype frequencies of VDR (Fok Ⅰ, Bsm Ⅰ, Apa Ⅰ, Taq Ⅰ) did not statistically differ between UC patients and the controls (all P>0.012 5). Stratification by the Truelove & Witts severity index, mutant allele (C) and genotype (TC+ CC) of VDR(Fok Ⅰ) were significantly higher in patients with mild and moderate UC than in those with severe UC (54.37%(373/686) vs 37.70%(46/122), 81.92%(281/343) vs 55.74%(34/61), bothP<0.01). Haplotype analysis showed that three polymorphic loci of Bsm Ⅰ, Apa Ⅰand Taq Ⅰwere in a complete linkage disequilibrium. The AAC haplotype decreased significantly in UC patients compared to the controls (3.58%(29/808) vs 6.01%(74/1 224),P=0.012). The average serum levels of 25(OH)D in UC patients were significantly lower than those in the controls ((48±17) vs (54±18)nmol/L, P=0.017). Furthermore, the average serum levels of 25(OH)D were significantly higher in patients with mild and moderate UC than in those with severe UC and were significantly lower in patients with extensive colitis than in those with distal colitis (both P<0.01). By linear regression analysis, the serum levels of 25(OH)D in UC patients were independently and positively correlated with hemoglobin (β=0.499, P<0.01) and yet independently and negatively correlated with C-reaction protein (β=-0.346, P<0.01) and white blood cells (β=-0.291, P=0.002). Using Logistic regression analysis, it was found that mutant genotype (GA/AA) of VDR (Bsm Ⅰ) played an independently protective role in UC (OR=0.328, P=0.028) while mutant genotype (TC/CC) of VDR (Fok Ⅰ) and vitamin D deficiency (<50.0 nmol/L) had an interaction in UC (OR=2.070, P=0.006).

Genetic polymorphism of VDR (Fok Ⅰ, Bsm Ⅰ, Apa Ⅰ, Taq Ⅰ) and serum levels of 25(OH)D are significantly correlated with UC. Mutation of VDR (Bsm Ⅰ) is a protective factor for UC. Moreover, mutant genotype (TC/CC) of VDR (Fok Ⅰ) and vitamin D deficiency may exert synergistic effects on the susceptibility to UC.

To explore the relationship between thyroid-stimulating hormone (TSH) and metabolic syndrome (MS) and its components in euthyroid male elders.

For this cross-sectional study, 307 euthyroid male elders aged over 60 years participating in a routine annual health screening examination at our hospital during May-June 2011 were enrolled. Their mean age was (72.6±7.9) years. Height, weight, blood pressure, blood glucose, blood lipids, fasting insulin levels and thyroid functions (TT_3,TT_4,FT_3,FT_4,TSH, TgAb & TPOAb) were measured. And the association of thyroid functions and the presence of MS and its components was analyzed.

The prevalence of MS was 40.7%(125/307). The TSH level was significantly higher in the MS and obeses group than that in control group ((2.6±1.1) vs (2.4±1.0)mU/L, P=0.014; (2.7±1.1) vs (2.3±1.1)mU/L, P=0.007). The prevalence of MS and obesity showed a gradual increase according to the TSH tertiles. When comparing subjects in the highest and lowest tertile of TSH, the former group demonstrated 1.872 and 1.904-fold increases in the odds ratio for obesity and MS after adjusting for age and homeostasis model assessment for insulin resistance (HOMA-IR) (95%CI:1.051-3.332, P=0.033; 95%CI:1.070-3.387, P=0.028). Logistic regression analysis showed that the TSH levels were independent influencing factors for MS in male elders (OR=1.324, 95%CI: 1.042-1.683).

High circulating TSH level, albeit normal, is associated with elevated risk for metabolic syndrome in male elders.

To explore the concentration changes of Tamm-Horsefall protein (THP) under centrifugation in rat urine and discuss its association with urolithiasis formation.

A total of 40 Wistar rats were divided randomly into 4 groups of flying with stone (A), flying without stone (B), stone without flying (C) and control (D). After centrifugation, the THP concentrations of each group were measured by enzyme-linked immunosorbent assay (ELISA). Then urinary system was dissected, stained with hematoxylin & eosin and observed under electron microscopy to examine the distribution and number of each section. The SPSS 13.0 software was used for data analyses.

Group A showed significant difference in THP concentrations with groups C and D ((11±4)vs(15±6), (17±4) ng/ml, P=0.037 and 0.005). No statistically significant difference existed between groups A and B ((11±5) ng/ml, P=0.998) or groups C and D (P=0.422). Group B had significant difference in THP concentrations with groups D (P=0.036). Regarding the number of stones in ureter, Group A had statistically significant difference with B (P=0.029). However, there was no difference in the number of bladder stones. In kidney stones, there was significant difference (P=0.029) on "+ + " rating.

Centrifugation may reduce the urinary concentration of THP so as cause uroliathiasis formation in rats.

To explore whether palmitate-induced apoptosis of osteoblastic MC3T3-E1 cell is mediated by an activation of nuclear factor-kappa B (NF-κB).

Cell viability was assessed with methyl thiazolyl tetrazolium (MTT) assay and cell apoptosis by Hochest 33258 staining. Palmitate was added at different timepoints and dosages. Western blot was used to evaluate the expression levels of IκBα, p-NF-κB p65 and NF-κB p65 protein.

Palmitate led to a dose- and time-dependent decreases in cell viability and increase in cell apoptosis. Cell viability dropped to 54% and cleaved caspase-3 increased 3.1-fold in cells treated with 500 μmol/L palmitate compared to control. The level of p-NF-κB p65 protein markedly increased at 60 min post-stimulation and reached a 2.96-fold increase of baseline level at 120 min( P<0.05). The IκBα level markedly declined at 60 min post-stimulation and decreased by 57% at 120 min(P<0.05). Compared to the group with palmitate treatment alone, pyrrolidinedithiocarbamic acid (10/20 μmol/L) significantly inhibited the palmitate-induced increase of p-NF-κB p65(1.39±0.12, 1.25±0.10 vs 1.76±0.14, bothP<0.05), restored the palmitate-induced decrease of caspase-3(2.24±0.28 vs 1.29±0.27,P<0.05)and inhibited the palmitate-induced increase of cleaved caspase-3(0.63±0.01 vs 1.13±0.10,P<0.05).

Palmitate induces apoptosis of MC3T3-E1 cell by an activation of NF-κB.

To compare the biological characteristics of epidermal stem cells (ESCs) from hypertrophic scar and normal skin.

Epidermal stem cells were separated, enriched and adhered from 20 hypertrophic scars (scar group) and 20 normal skins (control group). The morphology and growth characteristics of primary epidermal stem cells were observed. The absorbance (A) values of expression of Keratin 19, nucleoprotein p63 and integrin β ₁ were tested by Western blot. And the genes of Oct-4 and Nanog were tested by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Flow cytometry was used to examine the markers of integrin CD29, integrin CD49f and Keratin 19 of ESCs.

The primary cultured cells showed the similar shape and growth curve. By comparing the epidermal stem cells from scar groups and control groups, the absorbance values of the expression of Keratin 19, nucleoprotein p63 and integrin β ₁ were 860±4, 712±3, 422±6 and 862±3, 707±9, 413±6 (all P>0.05). The expression values of Oct-4, Nanog were (7.79±0.44)×10^-4,(5.96±0.36)×10^-4 and (7.93±0.29)×10^-4,(6.06±0.35)×10^-4 (all P>0.05). Percentages of positive cells expressing CD29, CD49f and Keratin 19 were (97.3±0.7)%, (94.6±1.1)%, (92.5±0.8)% and (98.8±4.6)%, (98.9±0.4)%, (94.4±0.7)% respectively (allP<0.05).

The ESCs in hypertrophic scar have the same characteristics with ESCs in normal skin. However, the ESCs from hypertrophic scar are lower than that from normal skin.

To explore the effects of adiponectin (APN) on high glucose-induced endoplasmic reticulum stress (ERS)-mediated apoptosis and cystoskeleton.

The conditionally immortal human glomeroular podocytes were divided into normal glucose, mannitol, high glucose and high glucose with adiponectin groups. Flow cytometry was employed to assess cell apoptosis. Real-time polymerase chain reaction (PCR) and Western blot were used to detect the expressions of such ERS molecules as GRP78, CHOP and caspase 12 and desmin and TRPC6. Immunofluorescent staining was used to detect the changes in the skeleton of podocyte.

(1) The apoptosis rate in high glucose group is significantly higher than the other groups (26.15%±1.38% vs 2.39%±0.58%, 4.84%±0.87%, 16.71%±1.15%, all P<0.05). Compared with control group, the mRNA and protein expressions of GRP78, CHOP and caspase 12 were all up-regulated significantly in high glucose group (P<0.05). The high glucose with adiponectin group could reduce the podocyte apoptosis by 10% and down-regulated the mRNA and protein expressions of GRP78, CHOP and caspase 12 versus high glucose group (allP<0.05). (2) High glucose-induced podocyte caused the up-regulated expressions of TRPC6 and desmin (allP<0.05) and it was inhibited by adiponectin (allP<0.05). Additionally, imunofluorescent assay of high glucose-induced podocyte cytoskeleton showed disorderly F-actin and absent tensile fiber. Adiponectin prevented the F-actin cytoskeleton disruption under high glucose.

Adiponectin plays a protective role in high glucose-induced podocyte through reducing endoplasmic reticulum stress-induced apoptosis and blunting the injury of cytoskeleton.

To explore the effect and mechanism of ryanodine receptor antagonist dantrolene on skeletal muscle of rats with severe scald injury.

A total of 56 Wistar rats were divided into control, scald and dantrolene treatment groups according to a random digital table. Rats in scald and dantrolene treatment groups were subject to 50% total body surface area (TBSA) full-thickness scald by a 12-second immersion of back and a 6-second immersion of abdomen in 94 ℃water and then received an intraperitoneal injection of Ringer's solution. At the same time, the rats in scald group received 5% mannitol through caudal vein while those in dantrolene treatment group received dantrolene 2 mg/kg (dissolved in 5% mannitol). Rats in control group were sham-injured through an immersion of back and abdomen into 37 ℃ warm water. Tibialis anterior muscle samples were harvested at Days 1, 4 and 7 post-scalding. Changes of skeletal muscle ultrastructure were observed by transmission electron microscope, subcellular calcium ion (Ca²⁺) contents of skeletal muscle (including cytoplasm, mitochondria & sarcoplasm reticulum) were detected by electron probe X-ray microanalysis (EPMA) and the levels of calpain-1 and calpain-2 protein were determined by Western blot. And the activities of calpain were detected by enzyme-linked immunosorbent assay.

In scald group, assorted arrangement appeared immediately at Day 1 post-injury and partial disappearance of Z lines at Day 7 post-injury. There were no significant ultrastructure changes in dantrolene treatment group at Day 1 and 4 post-injury. Curled filament and mild fracture occurred merely in dantrolene treatment group at Day 7 post-injury. The cytoplasmic contents of Ca²⁺ were significantly higher in scald group than those in control group at Day 1 and 4 ((0.964±0.060), (0.639±0.067) vs (0.266±0.029) μmol/L respectively, all P<0.05) while the contents of Ca²⁺ within sarcoplasm reticulum were obviously lower in scald group than those in control group at Day 1 and 4 ((0.368±0.060), (0.814±0.089) vs (1.337±0.112) μmol/L respectively, all P<0.05). However, those subcellular regions in dantrolene treatment group ((0.310±0.069), (0.490±0.039) and (1.241±0.073), (1.161±0.094) μmol/L) had no significant difference with control group (allP>0.05). Caplain-1 and calpain-2 protein levels in scald group increased significantly at Day 1 and 4 post-injury versus control group (1.371±0.034, 1.214±0.030 vs 0.838±0.017 & 1.464±0.015, 1.390±0.023 vs 0.806±0.026 respectively, allP<0.05), whereas caplain-1 and calpain-2 protein levels in dantrolene treatment (0.984±0.031, 0.935±0.023 and 0.836±0.014, 0.741±0.020) obviously were lower than those in scald group respectively (allP<0.05). The activities of calpain in scald and dantrolene treatment groups at Day 1, 4 and 7 post-injury were (8.33±0.21), (9.33±0.21), (10.59±0.18) and (7.76±0.28), (7.86±0.20), (7.91±0.22) μmol/L respectively while the activity of calpain in control group was (7.62±0.19) μmol/L. The activities of calpain in scald group were significantly higher than those in dantrolene treatment and control groups (allP<0.05) whereas the activities of calpain in dantrolene treatment group had no obvious change versus control group (allP>0.05).

Dantrolene offers significant protection from skeletal muscle tissue damage and minimizes the ultrastructural change of tibialis anterior muscle induced by severe scald injury. The mechanism is probably through inhibiting an excessive release of Ca²⁺ within sarcoplasm reticulum and down-regulated cytoplasmic expression and activity of calpain-1 and calpain-2.

Nasal endoscopy has been introduced in China since the 1980s. In recent years, endoscopic skull base surgery has become a hot spot, and it has been widely carried out, especially the technique of pituitary tumor surgery has become mature. However, the treatment of skull base tumors in children under nasal endoscopy is still rare. Six cases of pediatric skull base tumor resection under nasal endoscopy were jointly performed by the Department of Otolaryngology, Head and Neck Surgery and the Department of Neurosurgery of Tianjin Huanhu Hospital, which are reported as follows.

Colorectal cancer is one of the common malignant tumors in western countries. U.S. cancer statistics show that there are more than 100,000 new cases of colon cancer every year, ranking third among all malignant tumors in incidence and second among all tumors in mortality^[1]。 With the improvement of people's living standard, the change of dietary structure and the influence of environmental factors, the incidence of colorectal cancer in China has also increased obviously. According to the 2007 National Cancer Registry, colorectal cancer has risen to the third highest incidence rate and the fifth highest mortality rate among all malignancies^[2]。

C. difficile infection (Clostridium difficileinfection (CDI) is an infectious disease with the intestinal tract as the main manifestation caused by the excessive proliferation and release of toxins by the intestinal pathogenic Clostridium difficile. The clinical manifestations can range from mild self-healing diarrhea to severe pseudomembranous colitis, sepsis syndrome, toxic megacolon and even intestinal perforation^[1]。 Since the 21st century, the incidence and severity of CDI have been increasing worldwide, especially the emergence and outbreak of high-producing strain of Clostridium difficile (BI/NAP1/027 toxin type III), which makes Clostridium difficile one of the important pathogens related to medical and health^[2]。 Antibiotics have been used to treat CDI for more than 30 years, mainly metronidazole and vancomycin. However, in recent years, the sensitivity of these drugs has decreased, and the recurrence rate after cure is as high as 25% ~60%^[3]So that patients with severe CDI can only undergo subtotal colectomy after failure of antibiotic therapy. Therefore, the correct treatment of CDI and prevention of its recurrence is particularly important, and the development of new methods is imminent.

Recurrent hypoxemia (with or without hypercapnia) due to upper airway obstruction or stenosis during sleep in patients with obstructive sleep apnea syndrome (OSAS); Or sleep structural disorders due to repeated micro-arousals. Epidemiological data show that about 4% of men and 2% of women in adults suffer from OSAS^[1]。 OSAS can lead to pathophysiological changes such as chronic intermittent hypoxia, sleep fragmentation and sympathetic nerve excitation, which can lead to daytime drowsiness, cognitive dysfunction, mental and psychological abnormalities, etc., and can induce cardiovascular diseases such as hypertension, coronary atherosclerotic heart disease, arrhythmia, cardiac insufficiency or heart failure^[2]Cardiovascular comorbidities are the leading causes of disability and death in OSAS. The pathophysiological relationship between OSAS and cardiovascular diseases is complex and diverse. The related mechanisms and treatment progress are reviewed below.