[Analysis of clinical and pathological characteristics of high- risk HPV-negative carcinoma of the uterine cervix].

Objective: To study the clinical and pathological characteristics of HPV-negative cervical cancer patients. Methods: Retrospective analysis of 785 cervical cancer patients in Guangdong Women and Children Hospital from Jan. 2005 to Oct. 2015. By detecting high- risk HPV infection by flow-through hybridization genechip technique. Results: (1)Among 785 cases of cervical cancer, 71 cases were negative for HPV infection tested by genechip technique, accounting for 9.0%(71/785), and the relative light units/cut off(RLU/CO)ratios of these 71 cases were less than 1 by hybird capture Ⅱ(HC-Ⅱ)methods. The results showed that the positive coincident rate of genechip technique detecting result with HC-Ⅱ method was 100.0%(71/71).(2)There was no difference between 43(60.6%)cases from 41-55 years old of 71 cases of HPV-negative patients and 392(54.9%)cases from 41- 55 years old of 714 cases of HPV infection patients(χ2=15.63, P=0.571). Among 71 cases of HPV-negative patients, 32 cases of patients with doing TCT, 6(18.8%)cases for normal, 10(31.2%)cases for atypical squamous cells of undetermined significance(ASCUS), 3(9.4%)cases for atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion(ASC-H), 3(9.4%)cases for low-grade squamous intraepithelial lesions(LSIL), 8(25.0%)cases for hight-grade squamous intraepithelial lesions(HSIL), 2(6.2%)cases for squamous cell carcinoma(SCC). And there were 391 cases of patients with doing thin-prep cytologic test(TCT)of 714 HPV infection patients, 60(15.3%)cases for normal, 61(15.6%)cases for ASCUS, 28(7.2%)cases for ASC-H, 29(7.4%)cases for LSIL, 164(41.9%)cases for HSIL, 49(12.5%)cases for SCC. There was no difference of TCT between HPV infection and HPV-negative patients(P>0.05). Among 70 cases from 71 patients with negative for HPV infection, there were 8(11.4%)cases in stage Ⅰ a, 26(37.1%)cases in stage Ⅰ b1, 12(17.1%)cases in stage Ⅰb2, 11(15.7%)cases in stage Ⅱa, 10(14.3%)cases in stage Ⅱb, 3(4.3%)cases in stage Ⅲ-Ⅳ. There were 118(16.6%)cases in stage Ⅰa, 261(36.8%)cases in stage Ⅰb1, 72(10.1%)cases in stage Ⅰb2, 152(21.4%)cases in stage Ⅱa, 87(12.3%)cases in stage Ⅱb, 20(2.8%)cases in stage Ⅲ-Ⅳ in 710 cases of HPV infection patients, in which there were no difference of clinical stage between HPV infection and HPV-negative patients(P>0.05). Among 69 cases from 71 patients HPV-negative infection, there were 51(73.9%)cases for squamous carcinoma, 13(18.8%)cases for adenocarcinoma, 5(7.2%)cases for adenosquamous carcinoma; and 593(87.2%)cases for squamous carcinoma, 38(5.6%)cases for adenocarcinoma, 39(5.7%)cases for adenosquamous carcinoma, 10(1.5%)case for others were in 680 patients of HPV infection, in which there was significant difference of adenocarcinoma between HPV infection and HPV-negative patients(χ2=11.96, P=0.001). Conclusions: Flow-through hybridization genechip technique is the method of high sensitivity to detect high-risk type HPV, as like HC-Ⅱ method. HPV-negative of cervical cancer occurs mainly in 41- 55 years old. Adenocarcinoma incidence is significantly higher in HPV-negative cases than those patients with infection of HPV positive.

[Experimental study on mechanism of steroid-induced avascular necrosis of femoral head].

The authors adopted an animal model to observe the pathogenic mechanism of steroid-induced avascular necrosis of the femoral head. Sixty-four white rabbits were divided into two groups: hydrocortisone acetate (8 mg/kg) was hypodermically given to 48 experimental animals and 0.32 mg/kg of normal saline to 16 rabbits for control. Two groups of animals were fed and kept in the same condition. The results showed that application of the steroid drug could produce fat degeneration and necrosis of osteocytes and fat embolism in the small blood vessels of the femoral head. The abnormal hypertrophied fat cells in the bone marrow compressed small veins in the femoral head resulting in blood stasis of the capillaries, thus growth and regeneration of the capillary were inhibited.

Progesterone primes zona pellucida-induced activation of phospholipase A2 during acrosomal exocytosis in guinea pig spermatozoa.

We investigated, using guinea-pig spermatozoa as a model, whether phospholipase A2 (PLA2) is involved in progesterone or zona pellucida (ZP)-stimulated acrosomal exocytosis, if progesterone enhances ZP-induced activation of PLA2, and mechanisms underlying PLA2 regulation. Spermatozoa were capacitated and labeled in low Ca2+ medium with [14C]choline chloride or [14C]arachidonic acid, washed, and then exposed to millimolar Ca2+ and progesterone and/or ZP. Each agonist stimulated decrease of phosphatidylcholine (PC) and release of arachidonic acid and lysoPC, indicative of PLA2 activation. Aristolochic acid (a PLA2 inhibitor) abrogated lipid changes and exocytosis, indicating that these lipid changes are essential for exocytosis. Exposure of spermatozoa to submaximal concentrations of both progesterone and ZP resulted in a synergistic increase of arachidonic acid and lysoPC releases, and exocytosis, suggesting that, under natural conditions, both agonists interact to bring about acrosomal exocytosis. Progesterone-induced PLA2 activation appears to be mediated by a GABA(A)-like receptor, because bicuculline (a GABA(A) receptor antagonist) blocked arachidonic acid release and exocytosis. In agreement with this, GABA mimicked progesterone actions. ZP-induced activation of PLA2 seemed to be transduced via G(i) proteins because pertussis toxin blocked arachidonic acid release and acrosomal exocytosis. PLA2 may be regulated by PKC because progesterone- or ZP-induced release of arachidonic acid was blocked by the PKC inhibitors staurosporine or chelerythrine chloride. PLA2 could also be regulated by the cAMP-PKA pathway; inclusion of the PKA inhibitor 14-22 amide or H-89 led to a reduction in arachidonic acid release or exocytosis after progesterone or ZP. Taken together, these results suggest that PLA2 plays an essential role in progesterone or ZP-stimulated exocytosis with progesterone priming ZP action.

Zona pellucida induces activation of phospholipase A2 during acrosomal exocytosis in guinea pig spermatozoa.

Phospholipase A(2) (PLA(2)) is activated in spermatozoa in response to progesterone and Ca(2+) ionophores, but to our knowledge, no study has yet reported zona pellucida (ZP)-induced activation of PLA(2). We investigated whether PLA(2) is involved in ZP-stimulated acrosomal exocytosis, if Ca(2+) is required for activation of PLA(2), and signal transduction pathways modulating PLA(2) using guinea pig sperm as a model. Spermatozoa were capacitated and labeled in low-Ca(2+) medium with [(14)C]choline chloride or [(14)C]arachidonic acid and were then exposed to millimolar Ca(2+) and various reagents and stimulated with ZP. Precapacitated spermatozoa exposed to millimolar Ca(2+) and stimulated with ZP experienced increases in arachidonic acid (AA) and lysophosphatidylcholine (lysoPC) levels and a parallel decrease in phosphatidylcholine level; these changes are indicative of PLA(2) activation. Simulation with ZP also led to acrosomal exocytosis in a high proportion of spermatozoa. Lipid changes and exocytosis were prevented if spermatozoa were exposed to aristolochic acid, a PLA(2) inhibitor, before treatment with ZP. Stimulation with ZP in medium without added Ca(2+) or in medium with millimolar Ca(2+) and EGTA or La(3+) resulted in no lipid changes or exocytosis. Pretreatment with pertussis toxin, a G(i) protein inhibitor, before stimulation with ZP blocked the release of AA and lysoPC as well as acrosomal exocytosis. Exposure of spermatozoa to the diacylglycerol (DAG) kinase inhibitor R59022 before ZP stimulation led to a significant increase in generation of lysoPC and exocytosis. Taken together, these results indicate very strongly that PLA(2) plays an essential role in ZP-induced exocytosis in spermatozoa, that PLA(2) activation requires Ca(2+) internalization, and that PLA(2) activation is regulated by signal transduction pathways involving G proteins and DAG.