OBJECTIVE: To explore new hallmarks affecting the prognosis of patients with laryngeal squamous cell carcinoma (LSCC) via investigating the expression of CyclinE and p27 in LSCC tissues. METHOD: The expression of CyclinE and p27 was detected via Elivision immunohistochemical staining in 160 LSCC tissues and 20 normal laryngeal tissues (NLT). The relationship between CyclinE/ p27 and LSCC/ NLT was analyzed via Log-rank analysis. The relationship of CyclinE and p27 protein was statistically analyzed by spearman correlation analysis. The relationship between CyclinE/p27 and clinical-pathology-factors of patients with LSCC, such as age, gender, tumor site, diameter, differentiation, lymph node metastasis and PTNM stage were analyzed by Chi-square test. The relationship between clinical-pathology-factors, CyclinE, p27 and overall survival time of patients with LSCC was analyzed via Cox multiplicity and Kaplan-Meier survival analysis. A significant difference was recognized by P<0.05. RESULT: In LSCC the positive rates of CyclinE and p27 protein was 62.50% and 41.25% respectively (P<0.05). In NLT the positive rates of CyclinE and p27 protein was 35% and 70% respectively (P<0.05). The expression of CyclinE or p27 protein was closely correlated with lymph node metastasis, PTNM stage of patients with LSCC (P<0.05). The expression of CyclinE and p27 had no significant correlations with patients' gender, age and tumor site, diameter differentiation (P>0.05 for all). A negative correlation was found between the expression of CyclinE and p27 protein, r= -0.767(P<0.05). Kaplan-Meier survival analysis showed that the overall survival rate of patients with LSCC was 36.9% (P<0.05). The 5-year survival rate in positive group of CyclinE was 8%, in negative group was 80% (P<0.05). On the contrary, the 5-year survival rate of patients with LSCC in positive group of p27 protein was 77.27%, the rate was 5.32% in negative group (P<0.05). Cox multivariate regression analysis revealed that lymph node metastasis, PTNM stage, CyclinE and p27 were independent risk factors of prognosis for patients with LSCC. CONCLUSION It is the molecular basis underlying the development and invasion/ metastasis of LSCC that activation of CyclinE gene accompanying inactivation of p27 gene. It is very important of co-detecting CyclinE and p27 protein to predict the prognosis of patients with LSCC.
Carcinoma, Squamous Cell ; metabolism ; pathology ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Female ; Head and Neck Neoplasms ; metabolism ; pathology ; Humans ; Laryngeal Mucosa ; metabolism ; pathology ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Oncogene Proteins ; metabolism ; Prognosis ; Squamous Cell Carcinoma of Head and Neck ; Survival Rate

This study was purposed to construct the recombinant hF9 minigene and its stable nonsense mutant cell lines, and to investigate its significance. Minigene hF9 was cloned into the mammalian expression vector pCMV-Tag3B; a nonsense mutant containing a premature termination codon (PTC) in the 121(st) amino acid residue was obtained by PCR site-directed mutagenesis; minigene hF9 and nonsense mutant were respectively transfected into HepG2 cells with G418 treatment to get stable HepG2-WT and HepG2-N cell lines. The results confirmed that the minigene hF9 and nonsense mutant were constructed successfully. The gene of interest was amplified by RT-PCR from the stable cell lines, and the minigene hF9 was expressed in the stable cell lines. It is concluded that the recombinant hF9 minigene and its stable nonsense mutant cell lines are constructed successfully. The cell lines can be used to screen the drugs treating the nonsense mutation-caused hemophilia according to PTC read-through approaches.
Cell Line, Tumor ; Codon, Nonsense ; Factor IX ; genetics ; Genetic Vectors ; Hemophilia B ; genetics ; Humans ; Recombinant Proteins ; genetics

Objective:To compare the clinicopathological characteristics of main and accessory lesions in patients with synchronous multiple esophageal lesions (SMEL, i. e. early esophageal cancer and intraepithelial neoplasia) and to explore their correlation.Methods:Data of 80 patients with SMEL treated by endoscopic resection in The First Medical Center of Chinese PLA General Hospital from November 2006 to September 2019 were retrospectively analyzed, and the clinicopathological characteristics as well. The lesions were divided into main and accessory lesions, and their correlation in macroscopic type, lesion location, pathological type and invasion depth in 70 patients with double SMEL were investigated.Results:The age of 80 patients with SMEL was 61.3±8.32 years, more common in males (83.8%, 67/80). Fifty-seven patients (71.2%) had a history of smoking and drinking, respectively. There was a positive correlation between the size of main and accessory lesions in the 70 patients with double SMEL ( r=0.464, P<0.001). The macroscopic type ( P=0.115), location ( P=0.340) and depth of invasion ( P=0.555) of the main and accessory lesions were not correlated, but the pathological type had high correlation ( P<0.001). The consistency rate was 50.0% (35/70). Conclusion:Most SMEL patients are elderly males with a history of smoking and drinking. When one lesion is found, there is high possibility of multiple lesions. Physicians should be aware of the correlation between main and accessory lesions to avoid missed diagnosis.

OBJECTIVETo investigate the expression of several homeobox genes during the wound healing in fetal and adult skin and their roles in fetal scarless wound healing.

METHODSThe expressions of PRX-2, HOXB13, HOX2.2 and HOX2.3 during wound healing in fetal and adult skin were determined with in situ hybridization.

RESULTS(1) PRX-2 positive expression could be identified in normal fetal and adult skin, especially in the fetus. But there was difference in location sites of the genes. The positive expression in normal fetal skin was mainly found in the peripheral cells at the hair shafts within dermal papilla layers and was also found in the epithelium. Nevertheless, weak positive expression of PRX-2 was found in the epithelial basal layer cells in normal adult skin but not in dermal tissue. There was strong positive expression of the PRX-2 in the tissue around the wound in fetus but not of that in adults except the epithelial basal layers. (2) Positive expression of HOXB13 could be identified in both normal fetal and adult skins. And the expression was concentrated mainly in hair follicle cells in the dermis and in the basal layer cells in the epithelium. Furthermore, the expression became weak after trauma, especially in fetal skin. (3) The positive expression of HOX2.2 and HOX2.3 in normal fetal skin was observed mainly in the whole layer of the epithelium and especially in the epithelial basal layers. Weak positive expression could be found in the dermis and strong expression found in the tissue near the wound. But there was no positive expression of the HOX genes in normal adult skin and wounds.

CONCLUSIONThe difference in the HOX expression in fetal and adult skin wound healing might be the key factor leading to different wound healing. Homeobox genes might be closely related with the developmental biology.

Adolescent ; Adult ; Animals ; Burns ; pathology ; Child ; Female ; Fetus ; Gene Expression ; Genes, Homeobox ; Homeodomain Proteins ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Skin ; embryology ; Wound Healing ; genetics ; Young Adult

OBJECTIVETo explore the differences of PDGF and EGF expression in the wound healing between fatal and adult.

METHODSWith the established animal model of fetal scarless healing and the adult samples, an immunohistochemical technique was used to evaluate the expression of PDGF and EGF in the normal adult skin, normal fetal skin, and the process of their wound healing.

RESULTS1. The expression of the PDGF was not found in the fetal skin, but a mild amount of the PDGF was shown in the epidermis and the upper dermal layer 12 hours and 1 day after the wounding process. In the normal adult skin, expression of PDGF was shown in the dermal fibroblasts, macrophagocytes and blood capillaries, and a strong expression was presented during its wound healing process. 2. In the fetal skin, the expression of the EGF was seen in the epidermis, hair follicles, sebaceous glands and sweat glands, but there were no markedly changes during the wound healing. In the adult skin, a positive stain of the EGF was shown in the basal layer of the epidermis while the mild stain in hair follicles and sweat glands. The level of the expression became gradually decreasing with the time going in the wounded adult skin.

CONCLUSIONThe different expression of growth factors between fetal and adult skin in wound healing may be one of the important reasons that the fetal wound could produce scarless healing.

Adult ; Epidermal Growth Factor ; metabolism ; Epidermis ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Hair Follicle ; metabolism ; Humans ; Platelet-Derived Growth Factor ; metabolism ; Skin ; injuries ; metabolism ; Sweat Glands ; metabolism ; Wound Healing

OBJECTIVETo investigate the effect of reconstruction of lower eyelid ectropion with expanded flap.

METHODSFourty patients with lower eyelid ectropion were reconstructed using tissue expander. The volume of the smallest expander was 30 ml, and that of the biggest one was 150 ml. The expand time was from 2-months to 3-months, then advancement or transposition flaps were created and employed in the defected lesion where the scar was removed just in one operation.

RESULTSAll patients have been followed up for 2-year with satisfactory results and no recurrences was appearance.

CONCLUSIONSApplication of expander reasonable may get satisfactory result in reconstruction of lower eyelid ectropion. The incision in donor site is hidden and the symptom seldom recurs.

Adolescent ; Adult ; Blepharoplasty ; methods ; Child ; Ectropion ; surgery ; Humans ; Male ; Surgical Flaps ; Tissue Expansion Devices ; Young Adult

OBJECTIVEto investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.

METHODSfifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.

RESULTScompared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.

CONCLUSIONSin patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.

APACHE ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Prognosis ; Prospective Studies ; Sepsis ; drug therapy ; etiology ; immunology ; Somatostatin ; therapeutic use ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; blood

This study aims to find good markers for predicting the prognosis of patients with esophageal squamous cell carcinoma (ESCC). Vasculogenic mimicry (VM) and the expression of hypoxia inducible factor-lα (HIF-lα)/E-cad protein in ESCC were investigated by immunostaining. The association between VM, HIF-lα/E-cad and clinicopathologic characteristics and 5-year-survival rate of patients with ESCC was analyzed. A total of 160 ESCC specimens were involved in this study and 28 specimens of normal esophageal mucosa served as controls. VM channels were identified in 78 (48.75%) of the 160 ESCC specimens and none of the normal esophageal mucosa was found to have VM. The rates of high-expression of HIF-lα and E-cad in ESCC were 43.75% and 38.75%, while the rates in control were 17.86% and 71.43%, respectively (P<0.05 for all). VM and the expression levels of HIF-lα and E-cad were significantly related to lymph node metastasis, serosa infiltration, PTNM staging and 5-year-survival rates of patients with ESCC (P<0.05 for all). VM was positively correlated with HIF-lα but negatively with E-cad, and HIF-lα was negatively correlated with E-cad (P<0.001 for all). The 5-year-survival rate of patients with ESCC was 6.41% (5/78) in VM group and 65% (52/82) in non-VM group, 7.14% (5/70) in high HIF-lα expression group and 57.78% (52/90) in low HIF-lα expression group. Oppositely, the 5-year-survival rate in high E-cad expression group was 80.65% (50/62) and that in low E-cad expression group was 7.37% (7/98) (P<0.05 for all). Cox multifactor regression analysis indicated that lymph node metastasis, PTNM stage, VM and expression levels of HIF-lα and E-cad were independent risk factors of patients with ESCC (P<0.05 for all). Combined detection of VM, HIF-lα and E-cad plays an important role in predicting the invasion, metastasis and prognosis of patients with ESCC.
Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Cadherins ; metabolism ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Esophageal Neoplasms ; diagnosis ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Neovascularization, Pathologic ; diagnosis ; metabolism ; Prognosis ; Reproducibility of Results ; Sensitivity and Specificity ; Survival Rate

The purpose of this study was to construct 4 types of nonsense-mutated eukaryotic expression plasmids of fIX gene, using pcDNA3.1 plasmid containing fIX cDNA as template, and to identify, then to perform their expression in COS-7 cells. These stop mutants constructed by site-directed mutagenesis based on PCR, and further confirmed by DNA sequencing. COS-7 cells were transfected with either the wild-type or mutated fIX expression constructs, then the relative expression levels of fIX mRNA were detected by real time fluorescent quantitative PCR. The result showed that except the designed sites, there were no other nucleotide mutation in the sequences of four nonsense mutants. The results of real time PCR proved that the nonsense-mutated vectors can be effectively expressed in COS-7 cells. It is concluded that the nonsense-mutated eukaryotic expression vectors of fIX gene have been successfully constructed and can express in COS-7 cells, which provides the material basis for further researches on mechanism and treatment of FIX deficiency and the function defects caused by nonsense mutation.
Animals ; Base Sequence ; COS Cells ; Cercopithecus aethiops ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Factor IX ; genetics ; Genetic Vectors ; Transfection

Femoral neck fracture (FNF) in the elderly patients is currently a major health challenge worldwide, with excessive consumption of medical resources, high incidence of complications as well as suboptimal outcome and prognosis. Hip joint arthroplasty (HJA) has been the mainstream treatment for FNF in the elderly, but the conventional surgical approaches and techniques are still confronted with a series of bottlenecks such as dislocation, limp and limb length discrepancy. In recent years, direct anterior approach (DAA) for HJA (DAA-HJA) has been a major new choice in the field of joint replacement, which achieves improved clinical effectiveness of HJA in the treatment of elderly FNF, due to the fact that DAA approach involves the neuromuscular interface and accords with the idea of soft tissue retention and enhanced recovery after surgery. However, there is still a lack of unified understanding of standard technique and procedure of DAA-HJA in the treatment of elderly FNF. Therefore, relevant experts from the Hip Joint Group of Chinese Orthopedics Association of Chinese Medical Association, Youth Arthrology Group of Orthopedic Committee of PLA, Orthopedic Committee of Chongqing Medical Association, Branch of Orthopedic Surgeons of Chongqing Medical Doctor Association and Sport Medicine Committee of Chongqing Medical Association were organized to formulate the " Chinese expert consensus on the technical standard of direct anterior hip arthroplasty for elderly femoral neck fracture ( version 2023)" based on evidence-based medicine. This consensus mainly proposed 13 recommendations covering indications, surgical plans, prosthesis selections, surgical techniques and processes, and postoperative management of DAA-HJA in elderly patients with FNF, aiming to promote standardized, systematic and patient-specific diagnosis and treatment to improve the functional prognosis of the patients.