Volume 93  ·   Issue 02  ·  2013

To examine the protein and mRNA levels of interleukin-9 (IL-9) and the frequencies of CD4⁺IL-9⁺T-cells in peripheral blood of patients with systemic lupus erytbematosus (SLE) and explore the roles of double positive T cells and IL-9 in the pathogenesis of SLE and the effects of glucocorticoid.

Twenty-eight hospitalized SLE patients were recruited and 12 healthy volunteers selected as normal controls.The mRNA levels of IL-9 in peripheral blood were measured by real time-polymerase chain reaction (RT-PCR),plasma protein of IL-9 by enzyme-linked immunosorbent assay (ELISA) and frequencies of CD4⁺IL-9⁺T-cells by flow cytometry.And the differences between two groups and the effects of glucocorticoid were analyzed.

(1) The mRNA levels of IL-9 were significantly elevated in SLE patients as compared with normal controls (P<0.01).The serum levels of IL-9 were significantly higher in active and inactive SLE patients than those in healthy individuals (68±11 vs 26±6 ng/L,P<0.01:56±14 vs 26±6 ng/L,P<0.05).The percentages of CD4⁺IL-9⁺T-cells increased in active SLE patients (1.96%±0.31%) versus inactive SLE patients (0.89%±0.13%,P<0.01) and healthy controls (0.28%±0.05%,P<0.001).And it was higher in inactive SLE patients than that in controls (P<0.05). (2) The serum levels of IL-9 and the frequencies of CD4⁺IL-9⁺T-cells were positively correlated with SLE disease activity index (SLEDAI). (3) The frequencies of CD4⁺IL-9⁺T-cells and the serum levels of IL-9 in 8 untreated active SLE patients decreased at weeks 1,2 and 3 after the therapy of methylprednisolone (0.8 mg·kg^-1·d^-1) versus those at pre-treatment.

The abnormalities of IL-9 and CD4⁺IL-9⁺T-cells may play an important role in the pathogenesis of SLE.And the frequencies of CD4⁺IL-9⁺T-cells and the levels of IL-9 are evaluative parameters of SLE activity and severity.

To examine the clinical features of hypoparathyroidism (HP) with altered serum muscle enzymes and explore the correlations with age,gender,disease duration,serum total calcium,serum total phosphorus and parathyroid hormone (PTH).

From January 2005 to October 2011,a total of 41 HP patients at our hospital were retrospectively analyzed.

There were 16 patients with elevated serum muscle enzymes and 25 with normal serum muscle enzymes.The former group included 11 patients of idiopathic hypoparathyroidism (IHP) and 5 of pseudohypoparathyroidism (PHP) :serum calcium was (1.53±0.25) mmol/L and serum phosphorus (2.1±0.6) mmol/L:medians of creatine kinase (CK),lactic dehydrogenase (LDH),aspartate amino transfer enzymes (AST) were 309 (126-10 046),279 (131-741) and 27 (17-467) U/L in HP patients respectively.And the latter group included 20 IHP patients and 5 PHP ones:serum calcium was (1.64±0.29) mmol/L and serum phosphorus (2.2±0.6) mmol/L.The levels of serum muscle enzyme were compared before and after treatment in the former group.And they significantly declined with the rising serum level of calcium after treatment.The serum levels of CK (r=-0.82),LDH (r=-0.55) and AST (r=-0.56) in HP patients with elevated serum muscle enzymes had a negative correlation with the serum level of calcium (all P<0.05).And there was no correlation with age,gender,disease duration or serum total phosphorus (P>0.05).

Elevated serum muscle enzymes in HP patients have a negative correlation with serum level of calcium and there is no correlation with age,gender,disease duration,serum total phosphorus or parathyroid hormone.And elevated serum muscle enzymes are gradually restored with the normalized serum level of calcium.

To explore the effects of α-linolenic acid on insulin sensitivity in obese patients.

From October 2011 to April 2012,16 patients received an oral dose ofα-linolenic acid for 8 weeks.Oral glucose tolerance test (OGTT) and insulin releasing test were performed before and after treatment.Homeostasis model assessment insulin resistance (HOMA-IR) index and area under curve of insulin (AUCI) were calculated to evaluate the insulin sensitivity.The levels of serum triglyceride,free fatty acids (FFA),interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured after an overnight fast.

Obese patients had significantly elevated serum insulin,triglyceride,FFA,IL-6 and TNF-αversus the subjects in normal control group (all P<0.05).Obese patients were also more insulin-resistant than normal subjects based on a higher HOMA-IR (P<0.05).Decreased serum insulin,triglyceride,FFA,IL-6 and TNF-αwere observed after treatment.With the administration ofα-linolenic acid,HOMA-IR and AUCI significantly decreased in obese patients (HOMA-IR:1.8±0.2 vs 1.2±0.3,P<.05:AUCI:1151±505 vs 768±347,P<0.05).

α-linolenic acid increases peripheral insulin sensitivity in obese patients and it may aid the prevention and treatment of type 2 diabetes mellitus and atherosclerotic vascular diseases.

To explore the feasibility and clinical efficacies of unbilical one-trocar laparoendoscopic surgery versus trans-umbilicus and abdominal wall two-trocar laparoendoscopic surgery in the treatment of pediatric hydrocele.

Retrospective comparative analysis was conducted for 78 cases of hydrocele undergoing laparoscopic surgery at our hospital from January 2012 to May 2012.They were divided into two groups of umbilical one-trocar laparoscopic surgery (one-trocar,n=32) and trans-umbilicus and abdominal wall two-trocar laparoscopic surgery (two-trocar,n=46).And their profiles of operative duration,post-operative hospital stay and treatment cost were compared.

All procedures were successful.No case converted into open surgery.Visual field of both methods was similar,but two-trocar group had a flexible visual angle.During a follow-up period of 3-6 months,there was no occurrence of postoperative complications.The average operative duration was (20±10) min at one side and (31±11) min at both sides in one-trocar group versus (20±8) min and (29±9) min in two-trocar group.There were no statistical significance (all P>0.05).Cost in one-trocar group was (5199±599) yuan RM Band (5117±684) yuan RM Bin two-trocar group (P>0.05).

Trans-umbilicus laparoendoscopic one-trocar surgery is both feasible and safe in the treatment of pediatric hydrocele.Compared with two-trocar laparoscopic surgery,both approaches are similar in terms of operative duration,post-operative hospital stay and treatment cost.Since there is a single hidden navel scar,the former is labor-saving,may be handled by one operator and offers better cosmetic outcomes.

To determine the impact on tyrosinase expression and export from endoplasmic reticulum by inhibition of 26S proteasome.

Western blot was used to detect 26S proteasome from 8 vitiligo patients and 4 healthy controls.Melanocytes were incubated with proteasome inhibitor (lactacystin) and further detected as follows:cell survival by MTT assay,proteasome activity with fluorescence,ultrastructure observation with electron microscope,co-localization of tyrosinase and calreticulin (endoplasmic reticulum marker) by confocal laser scanning microscopy and 26S proteasome and tyrosinase with Western blot.

The 26S proteasome expression level from lesions of vitiligo (1.05±0.40) was significantly lower than the donor sites (1.82±0.88) and the healthy controls (1.88±0.16) (P<0.05).But no significant difference existed between the latter two groups (P>0.05).Compared to the untreated group,a 12-h incubation of 10μmol/L lactacystin showed inhibitory effects on melanocytes (0.999±0.110 vs 1.372±0.127,P<0.05) and significantly decreased proteasome activity (0.234±0.019 vs 1,P<0.01).Expansion rate of endoplasmic reticulum in the lactacystin group (1.91±0.17) was significantly higher than that of the untreated cells (1.17±0.11) (P<0.05).More tyrosinase co-localized with calreticulin in endoplasmic reticulum in lactacystin-treated cells was observed than that of the untreated group.Compared with the untreated group,significantly decreased levels of tyrosinase (146±10 vs 269±8,P<0.01) and tyrosinase activity (0.159±0.017 vs 0.221±0.019,P<0.01) were shown in the lactacystin group (P<0.05).

Significantly decrease of 26S proteasome is found in lesions of vitiligo patients.Inhibition of 26S proteasome may lead to expansion of endoplasmic reticulum of melanocytes,impact export of tyrosinase from melanocyte endoplasmic reticulum and expression of tyrosinase.

To explore the features of coronary atherosclerosis among the patients at different ages and examine their risk factors and pathological characteristics.

Retrospective analysis was performed for the clinicopathologic data of 87 patients with coronary atherosclerosis confirmed through autopsy from February 1986 to December 2011 at our hospital.They were divided into 3 groups according to age (20-<40 years,n=12:40-<60 years,n=24:≥60 years,n=51).Comparative analysis was performed for the degree of coronary artery stenosis and cardiac pathological changes,vulnerable plaque occurrence and morphological features of acute coronary syndrome among different patient groups.

There were 56 males and 31 females with a mean age of 66.8 years (range:23-80).Great statistical differences existed in the degree of coronary artery stenosis between different genders (P<0.05) while there was no significant difference among the above age groups (P>0.05).In the above age groups of patients with acute coronary syndrome,the incidence of big lipid core was 23.5%,38.5% and 53.8% (χ²=6.282,P=0.043),that of thin fibrous cap 29.4%,41.0% and 58.8% (χ²=6.589,P=0.037),that of inflammatory cell in 2 filtration 58.8%,69.2% and 85.0% (χ²=7.435,P=0.024) and that of calcification formation 35.3%,56.4% and 71.3% respectively (χ²=8.599,P=0.014).And the incidence of vulnerable plaque occurrence was 76.5%,84.6% and 88.8% (χ²=1.850,P=0.397) in the above age groups.

The features and risk factors of coronary atherosclerosis are different according to different ages and genders.Thus we should pay more attention to the early diagnosis and reasonable treatments of young patients and those with both mild coronary stenosis and vulnerable plaque.

To evaluate the efficacy and prognostic factors of autologous hematopoietic stem cell transplantation (ASCT) in multiple myeloma (MM) patients.

Retrospective analysis was performed in 27 MM patients undergoing ASCT at our hospital from May 2004 to August 2011.After comparing with 28 patients achieving very good partial response (VGPR) or better outcome and not undergoing ASCT,the impact on the extent of response,progression-free survival (PFS) and overall survival (OS) as well as related prognostic factors of MM patients were analyzed.

All patients successfully underwent hematopoietic reconstruction without transplantation-related mortality.The complete remission (CR) rate of ASCT group increased from 25.9% (7/27) at pre-ASCT to 70.4% (19/27) at post-ASCT (P<0.01).The estimated 5-year rate of progression-free survival was 56.2% (median not reached) in the ASCT group and 24.9% (median 29 months) in the non-ASCT group (P<0.05).The 5-year probability of overall survival was 52.2% (median not reached) in the ASCT group and 33.1% (median 60 months) in the non-ASCT group (P>0.05).Univariate analysis in ASCT group demonstrated that maintenance/consolidation therapy was associated with PFS (P=0.010) and OS (P=0.008).Patients on induction therapy containing boretizomib and early ASCT maintenance therapy all survived without disease progression until final follow-up (P=0.010).

ASCT can further increase the CR rate,prolong PFS and probably OS.The incorporation of novel agents into induction,consolidation and maintenance phases has optimized the anti-myeloma activity of ASCT and may be important for improved long-term outcomes.

To assess the clinical outcomes of transaortic extended septal myectomy on early and midterm survival of patients with hypertrophic obstructive cardiomyopathy (HOCM).

From October 2009 to April 2012,118 consecutive patients underwent extended Morrow′s procedure for HOCM.There were 69 males and 49 females with an average age of (46±13) years.Their clinical data were analyzed retrospectively.Preoperative transthoracic,intraoperative transoesophageal and postoperative transthoracic echocardiography was performed to assess septal thickness,left ventricular outflow tract (LVOT) gradient,mitral valve function and systolic anterior motion (SAM) of anterior mitral valve leaflet,etc.Concomitant surgical procedures were performed if other cardiac diseases required surgical interventions.Follow-up study was carried out during subsequent clinic visits at outpatient department and through telephone interviews with patients and their relatives.

The septal thickness was (25±7) mm.SAM was detected in all.The in-hospital mortality was 0.8% (1/118) since one patient died of multiple organs failure one week later.Postoperative echocardiography demonstrated marked reduction in LVOT gradient (92±22 vs 13±10 mm Hg (1 mm Hg=0.133 kPa),P=0.000),New York Heart Association (NYHA) class (2.9±0.6 vs 1.2±0.4,P=0.000) and significant improvement in mitral regurgitation.Concomitant surgical procedures were performed in 45 cases (38.1%,all for preexisting conditions).Complications included complete atrioventricular block (n=3),first degree atrioventricular block (n=6),complete left bundle branch block (n=51),intraventricular conduction delay or left anterior division block (n=26),transient renal dysfunction (n=2) and intra-aortic-balloon-pumping (n=2).No other severe complication was observed.During a follow-up period of 1-27 (7±6) months,there was no readmission or death.All patients reported significant increase in physical ability and obvious decrease in limiting symptoms.At the latest follow-up,the NYHA functional class maintained gradeⅠ-Ⅱin all.And mitral regurgitation remained absent or mild.

Surgical procedure for HOCM patients is both safe and efficacious.It provides an excellent relief of LVOT obstruction.

To explore the glycemic control status and related risk factors of overweight or obesity patients with type 2 diabetes mellitus (T2DM) in Guangdong province.

The medical records of overweight or obesity patients with T2DM from 60 tertiary and secondary hospitals in Guangdong Province were collected by questionnaire and physical examination.And the clinical data were analyzed to explore the influencing factors of glycemic control.The Hb A1c level was used to assess glycemic control.Hb A1c<7.0% indicated that glycemic control was up to standard.

From August 2011 to March 2012,5241 T2DM patients were recruited.The scope of current analysis was restricted to 4768 subjects with true data and deficiency no more than 5%.There were 2252 males and 2516 females.The age range was from 16 to 90 years,a median age 59.0 (50.0-69.0) years,onset age of diabetes 52.0 (44.0-60.0) years:a range of disease duration from 1 day to 42 years and a median of 5.0 (2.0-11.0) years.The median body mass index was 26.33 (24.88-28.34) kg/m²and median waist circumference 93.0 (88.0-100.0) cm.Median Hb A1c was 8.1% (6.9%-10.1%) and only 26.2% patients reached the target level of HbA1c<7.0%.Influencing factors of poor glycemic control were central obesity,high levels of resting heart rate,concurrent fatty liver and high intensity of treatment.And influencing factors of good glycemic control were regular exercises,smoking cessation,regular glycemic monitoring and good control of total chloestrol/triglyceride.

A majority of Guangdong type 2 diabetics fail to achieve target values for glycemic control.There is an urgent need for comprehensive management for improving glycemic control.

To explore the in vivo anticancer effects of luteolin with BGC-823 gastric carcinoma xenografts in nude mice and elucidate its mechanism.

After modeling of gastric carcinoma xenografts in nude mice,40 BALB/c (nu/nu) nude mice were randomly divided into 5 groups (n=8 each).And an intraperitoneal injection of luteolin was administered at 10 mg/kg (low-dose),20 mg/kg (middle-dose) and 40 mg/kg (high-dose) groups.And 5-fluorouracil (30 mg/kg) and control groups were also established.The growth curves of xenografts in nude mice were drawn and weight inhibition rates measured. The morphological features were detected by hematoxylin and eosin staining.And the protein expression levels of vascular endothelial growth factor A (VEGF-A) and matrix metalloproteinase 9 (MMP-9) were measured by immunohistochemistry.

In vivo tumor formation test showed that tumor volume in nude mice treated with luteolin was smaller than that of control group.Tumor weights of high-dose luteoin group were lighter than those of the control ((0.29±0.01) vs (0.38±0.03) g).And the difference was statistically significant (P<0.01).The rate of tumor inhibition in high-dose luteoin group was up to 24.87%.Lymphocytic invasion of tumor tissue was observed under light microscope in the treatment groups.Results of immunohistochemistry showed the positive cell integral of VEGF in middle and high-dose luteoin groups were 1.25±0.17 and 1.00±0.07 respectively.Both were significantly lower than that of control group (1.50±0.15,both P<0.05).The positive cell integral of MMP-9 in high-dose luteoin group was markedly lower than that of control group (3.75±1.43 vs 9.00±1.08,P<0.01).

Luteolin can effectively inhibit the in vivo growth of gastric tumor.The mechanism may be correlated with the stimulation of immune response and the down-regulated expressions of VEGF-A and MMP-9.

To explore the effects of tumor necrosis factor-alpha (TNF-α) or methylprednisolone on the expression of interleukin-17 receptor C (IL-17RC) in rat pulmonary microvascular endothelial cells (RPMVEC).

Culture RPMVEC were randomly divided into dose-dependent and time-dependent groups.In dose-dependent group,cells were cultured with TNF-α (0,0.1,1,10μg/L TNF-α) for 3 h.In time-dependent group,cells were cultured with TNF-α (10μg/L) for 0,1,3,6,12 h.In the drug intervention group,cells were cultured with TNF-α (10μg/L) or methylprednisolone (200μg/L) +TNF-α (10μg/L) for 3 h respectively.The expression of IL-17RC in isolated and cultured RPMVEC was identified by reverse transcription-polymerase chain reaction (RT-PCR),Western blot and immunocytochemistry.The changes of IL-17RC mRNA were detected by RT-PCR after the stimulation of RPMVEC by TNF-αor methylprednisolone.

RT-PCR and Western blot revealed that IL-17RC mRNA and protein were present in RPMVEC.The product of IL-17RC immunocytochemical reaction was predominantly located in cytoplasm and cytomembrane.In RPMVEC TNF-αsignificantly up-regulated IL-17RC mRNA in a dose-dependent manner (0μg/L TNF-αgroup:0.241±0.010,0.1μg/L TNF-αgroup:0.372±0.017,1μg/L TNF-αgroup:0.452±0.017,10μg/L TNF-αgroup:0.643±0.042,F=33.774,P<0.05).In time-dependent group,the expression of IL-17RC mRNA rose at 1 h (0.417±0.038),peaked at 3 h (0.674±0.018),then decreased gradually at 6 h (0.378±0.035),but stayed higher at 12 h (0.318±0.032).When compared with 0 h group (0.197±0.008),there were significant differences (F=37.903,P<0.05).Methylprednisolone caused a marked attenuation of TNF-α-induced IL-17RC expression (0.333±0.031 vs 0.660±0.026,F=89.637,P<0.05).

IL-17RC is predominantly present in cytomembrane and cytoplasm of RPMVEC.TNF-α up-regulates the expression of IL-17RC mRNA.Methylprednisolone inhibits the elevated expression of IL-17RC mRNA induced by TNF-α so as to relieve the inflammatory response of PMVEC.

To observe the preliminary efficacies and adverse events of sunitinib in the treatment of metastatic breast cancer ulcer.

From December 2008 to May 2010,patients with advanced breast cancer ulcer took a single sunitinib.The dosage was adjusted on the basis of adverse events.And clinical response was evaluated.

Nine patients with advanced breast cancer ulcer finished the treatment.The objective response and the clinical benefit time to progression of sunitinib were 3 and 7 patients with metastatic breast cancer ulcer,and the median time to progression (TTP) was 2.0 months.The most common adverse events included fatigue,hand-foot syndrome,neutropenia,thrombocytopenia and hypertension.

Single-agent sunitinib treatment of refractory advanced breast cancer ulcer has marked efficacies.However,neutropenia,thrombocytopenia and hypertension are the major dose-limited toxicities.

To study the value of a combination of vinorelbine and cisplatin (NP) as second-line neoadjuvant chemotherapy regimen for primary breast cancer.

Primary breast cancer patients on neoadjuvant chemotherapy and non-responsive to anthracyclines plus taxans received the NP regimen.The clinical objective response was evaluated with dynamic contrast-enhanced magnetic resonance imaging (MRI) according to RECIST 1.1 before operation.The pathological response was evaluated by the Miller-Payne grading system.And the toxicities were observed and evaluated according to National Cancer Institute-Common Terminology Criteria Version 3.0 (NCI-CTC v3.0).

A total of 33 breast cancer patients were examined.The outcomes were complete remission (CR,n=0,0%),partial remission (PR,n=16,48.5%),stable disease (n=17,51.5%) and progressive disease (n=0,0%).The clinical responsive rate (CR+PR) rate was 48.5%.The pathological response rates were G1 (n=6,18.2%),G2 (n=6,18.2%),G3 (n=10,30.3%),G4 (n=9,27.2%) and G5 (n=2,6.1%).And the pathological response (G3+G4+G5) was found in 21 cases (63.6%).The most common toxicities included neutropenia and nausea/vomiting.No serious toxicities were observed.

As a well-tolerated and effective regimen,NP regimen may be recommended as an option of second-line neoadjuvant chemotherapy regimen for primary breast cancer.

To explore the effects and underlying mechanisms of high glucose on in vitro invasiveness of human breast cancer cell line MDA-MB-435.

The invasiveness of MDA-MB-435 was determined by Matrigel-coated transwell chambers.Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were employed to analyze the cellular expression of matrix metalloproteinase-9/matrix metalloproteinase-2/E-cadherin (MMP-9/MMP-2/E-cadherin) gene/protein.

The invasive breast cancer cell numbers of each group (Glu 5.5,11 and 25 mmol/L) were 50±5,65±6 and 77±3 respectively.Cellular invasion was dramatically enhanced in the Glu 11 and 25 mmol/L group compared with the 5.5 mmol/L group.The MMP-9/MMP-2 protein expression increased significantly in the Glu 11 and 25 mmol/L groups compared with 5.5 mmol/L group while high glucose (Glu 11 and 25 mmol/L group) down-regulated significantly the E-cadherin m RNA/protein expression.

High glucose can promote the in vitro invasiveness of human breast cancer cells through the altered expression of MMP-9/MMP-2/E-cadherin.

To explore the effects of polysomy 17 on human epidermal growth factor receptor-2 (HER-2) testing and study its clinicopathologic significance.

We retrospectively analyzed the HER-2 status by fluorescence in situ hybridization (FISH) and HER-2 protein expression by immunohistochemistry (IHC) in a cohort of 619 patients with invasive breast cancer.The relationship between polysomy 17 and various clinicopathologic parameters was assessed.

Polysomy 17 was observed in 31.8% of cases,but more frequently in the IHC (3+)(41.9%) than in the IHC (2+) (27.7%) and IHC (1+/0)(11.1%) subgroup (P=0.001).There was no significant correlation between the frequency of polysomy 17 and HER-2 status in each IHC subgroup (P>0.05).Among the cases without gene amplification by FISH,9 of 15 IHC (3+) cases showed polysomy 17.As compared with the amplified group,unamplified polysomy 17 patients were associated with such good prognostic indicators as greater hormone receptor positivity (P<0.001) and lower Ki-67 index (P=0.003) with a trend towards those with neither amplification or polysomy.

The frequency of polysomy 17 is partially correlated with HER-2 protein expression but not HER-2 amplification.And polysomy 17 is a major factor in strong HER-2 protein overexpression in 3+nonamplified cases.Tumors displaying unamplified polysomy 17 resemble more HER-2-negative than HER-2-positive counterparts.