[Prognostic evaluation of nutritional indicators in patients with limited-stage small cell lung cancer].

Objective: To explore the effect of nutritional status pre-and during chemoradiotherapy on the prognosis of patients with limited- stage small cell lung cancer (LS-SCLC). Methods: We retrospectively collected medical records of 172 LS-SCLC patients undergoing concurrent chemoradiotherapy in our hospital from 2000 to 2014, with 126 males and 46 females. The data of complete blood count and hepatic and renal function were collected before initial treatment, before radiotherapy, 4 weeks during radiotherapy, and 1 month after complete of treatment. The prognostic nutritional index(PNI)was calculated. Kaplan-Meier method was used to calculate the survival rate. Log-rank test was performed used to compare the survival differences between groups. Multivariate prognostic analysis was performed using Cox regression model. Results: The median overall survival (OS) was 21 months, with median progression-free survival (PFS) of 11 months. At the beginning of treatment, patients with pre-treatment PNI ≥ 53 had significantly superior OS (median 37 vs 15 months, P=0.001) and PFS (median 16 vs 10 months, P=0.017). Patients with pre-treatment hemoglobin ≥140 g/L and <140 g/L had an median OS of 32 months and 17 months (P=0.019), and median PFS of 16 months and 9 months (P=0.040), respectively. During chemoradiation, patients with elevated hemoglobin had similar median OS compared with those had decreased hemoglobin (27 vs 18 months, P=0.063, but superior median PFS (15 vs 9 months, P=0.017). Multivariate analysis revealed that prophylactic cranial irradiation, pre-treatment hemoglobin ≥140 g/L, and pretreatment PNI ≥53 were independent predictors of OS and PFS in patients with LS-SCLC. Conclusion: Pre-treatment nutritional status and the changes of nutritional status during chemoradiotherapy is significantly associated with the prognosis of patients with limited-stage small cell lung cancer. The patients with better pre-treatment nutritional status have a better prognosis.

[Analysis on cancer incidence and mortality attributed to human papillomavirus infection in China, 2016].

Objective: We aim to evaluate the morbidity and mortality of cancer attributable to human papillomavirus (HPV) infection in China in 2016. Methods: Based on the cancer incidence and mortality rates, national population data, and population attributable fraction (PAF) in China, we calculated the number of incidence and death cases attributed to HPV infection in different areas, age groups, and gender in China in 2016. The standardized incidence and mortality rates for cancer attributed to HPV infection were calculated by using Segi's population. Results: In 2016, a total of 124 772 new cancer cases (6.32 per 100 000) were attributed to HPV infection in China, including 117 118 cases in women and 7 654 cases in men. Of these cancers, cervical cancer was the most common one, followed by anal cancer, oropharyngeal cancer, penile cancer, vaginal cancer, laryngeal cancer, oral cancer, and vulvar cancer. A total of 41 282 (2.03 per 100 000) deaths were attributed to HPV infection, of which 37 417 occurred in women and 3 865 in men. Most deaths were caused by cervical cancer, followed by anal cancer, oropharyngeal cancer, penile cancer, laryngeal cancer, vaginal cancer, oral cancer, and vulvar cancer. The incidence and mortality rates of cervical cancer increased rapidly with age, peaked in age group 50-54 years, then decreased obviously. The morbidity and mortality rates of non-cervical cancer increased with age. The cancer case and death numbers in rural areas (57 089 cases and 19 485 deaths) were lower than those in urban areas (67 683 cases and 21 797 deaths). However, the age-standardized incidence rate (ASIR) and age-standardized mortality rate (ASMR) of cervical cancer were higher in rural areas than in urban areas. There were no significant differences in ASIR and ASMR of non-cervical cancers between urban areas and rural areas. Conclusions: The incidence of cancers attributed to HPV infection in China was lower than the global average, but the number of incidences accounted largely, furthermore there is an increasing trend of morbidity and mortality. The preventions and controls of cervical cancer and male anal cancer are essential to contain the increases in cancer cases and deaths attributed to HPV infection.

Tear function index. A new measure of dry eye.

OBJECTIVE: To develop an efficient way to evaluate tear dynamics clinically. PARTICIPANTS: Three hundred fifty-two patients with dry eye, 64 of whom had Sjögren syndrome, and 55 normal subjects. DESIGN: Because various forces that affect tear drainage are reflected in the values of the Schirmer test with anesthesia and the tear clearance rate, we introduced a new measure of tear dynamics, the tear function index, which is the value obtained from dividing the value of the Schirmer test with anesthesia by the tear clearance rate. RESULTS: The tear function index was more specific (91.8%) and sensitive (78.9%) in diagnosing dry eye associated with Sjögren syndrome than was the Schirmer or tear clearance rate test alone. Tear function indexes below 96 were consistent with dry eye and those below 34 were seen primarily in patients with Sjögren syndrome. CONCLUSIONS: The tear function index offers a new method to evaluate tear production with the Schirmer test, introduces an extended way to measure tear flow combining with tear drainage, and gives a practical measure to diagnose dry eye.

Corneal organ culture model for assessing epithelial responses to surfactants.

The main goal of the present study was to investigate the response of cultured bovine corneas to the application of irritant substances and its potential use for predicting ocular irritancy in humans. We hypothesized that chemicals causing eye irritation may induce disruption of epithelial tight junctions and trigger cell stress responses modulated via transcription factors such as AP-1 and NF-kappaB. A simple air-lifted corneal organ culture system was used as an ex vivo model for ocular irritancy test. The effects of two surfactants, sodium dodecyl sulfate (SDS) and benzalkonium chloride (BAK), on corneal epithelial permeability and DNA-binding activity of AP-1 and NF-kappaB were studied in cultured bovine corneas. Both SDS and BAK induced tight junction disruption and increased permeability of corneal epithelium assessed using surface biotinylation in a concentration- and time-dependent manner. An increase in DNA-binding activity measured using electrophoretic mobility shift assay was observed when cultured corneas were treated with surfactants at concentrations causing minimal to mild ocular irritation, indicating epithelial cell stress response. Furthermore, exposure of cultured corneas to SDS or BAK at concentrations causing severe ocular irritancy resulted in a decrease in DNA-binding activity of these transcription factors in epithelial cells. These results indicate that the combination of corneal organ culture and measurements of corneal epithelial permeability and DNA-binding activity of stress-response transcription factors following chemical exposure has the potential to be used as a mechanistically based alternative to in vivo animal testing.

Correlation of tear clearance rate and fluorophotometric assessment of tear turnover.

BACKGROUND: The study sought to determine dynamic changes and theoretical bases of a clinical diagnostic test, the tear clearance rate. METHODS: Thirty four healthy subjects ranging in age from 22 to 84 years underwent examination of tear clearance rate, the Schirmer test with anaesthesia, as well as fluorophotometric measurement of tear turnover, tear volume, and tear flow. By applying 0.5% fluorescein into the conjunctival sac and subsequently measuring colour fades on a Schirmer strip, the tear clearance rate for assessing tear drainage was divided into nine grades. The results of the tear clearance rate were compared with those of the basal tear turnover and tear flow obtained from fluorophotometry. RESULTS: Significant relations were found between the tear clearance rate and the basal tear turnover or tear flow (r = 0.91 and 0.79, respectively, p = 0.0001). Considering the grades of progression from low to high, each grade of tear clearance rate showed a 12.5% increase in basal tear turnover (3.59%/min) and tear flow (0.38 microliter/min). There was no significant correlation between age and the basal tear turnover, tear volume, tear flow, or the tear clearance rate. CONCLUSION: The tear clearance rate is proposed as a simple and useful way to estimate basal tear turnover and tear flow, and measure tear drainage indirectly.

Immunohistochemical evidence of neuronal and glial differentiation in retinoblastoma.

BACKGROUND: The study sought to investigate the histogenesis of retinoblastoma. METHODS: One hundred specimens of retinoblastomas were examined along with those of 18 astrocytic gliomas and 15 medulloblastomas to compare similarities of glial differentiation in retinoblastoma and the two types of brain tumour. Employing avidin-biotin immunoperoxidase technique, antibodies were applied against neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP), and S-100 protein (S-100). RESULTS: Most rosettes and fleurettes, and some undifferentiated cells in retinoblastomas were NSE positive, but GFAP and S-100 negative. GFAP and S-100 positive cells in retinoblastomas were detected mostly in well differentiated glial cells which were interpreted as reactive or non-neoplastic cells. Some of the GFAP and S-100 positive cells in retinoblastomas were defined as tumour cells that resembled neoplastic astrocytes in astrocytic gliomas and medulloblastomas. CONCLUSION: Retinoblastoma may arise from primitive bipotential or multipotential cells capable of neuronal and glial differentiation.

Decrease in corneal sensitivity and change in tear function in dry eye.

Our purpose was to investigate the relationship between insufficient tear secretion and decreased corneal sensitivity. We studied 59 patients with dry eye, 15 of whom had Sjögren's syndrome (SS), and 26 healthy subjects. Corneal sensitivity was measured by the esthesiometer of Cochet and Bonnet. Schirmer test with or without anesthesia, tear clearance rate, tear function index, and rose bengal and fluorescein staining were also evaluated. The mean corneal sensitivity of either dry-eye group (4.6 +/- 1.2 and 4.5 +/- 1.2 cm for non-SS and SS dry eye, respectively) was significantly lower than that of the control (5.8 +/- 0.4 cm, p < 0.001). Corneal sensitivity correlated significantly with the Schirmer values with anesthesia and the tear function index in the two dry-eye groups and the control (p < 0.05). There were significant relationships between corneal sensitivity and the rose bengal and fluorescein scores in the three groups (p < 0.05). Hyposecretion of tears in dry eye may lead to pathologic changes in corneal epithelium and a decline in corneal sensitivity. Prompt treatment of dry eye is essential to maintain a normal corneal protective mechanism.

Survival of rabbit limbal stem cell allografts after administration of cyclosporin A.

OBJECTIVES: To evaluate the efficacy of systemic or topical administration of cyclosporin A (CsA) after limbal transplantation of stem cell allografts in rabbits. METHODS: Thirty-six rabbits underwent corneal epithelial debridement and limbal ablation to induce ocular surface disease and were then treated by limbal allograft transplantation. Animals received either systemic CsA (10 mg/kg per day, intramuscularly), 1% CsA eyedrops, or vehicle eyedrops immediately after transplantation and 28 days thereafter. Concentration of CsA in plasma and aqueous humor was determined by fluorescence polarization immunoassay after 4 weeks of therapy. Graft survival was inspected clinically. RESULTS: Both systemic and topical administration of CsA resulted in a significant prolongation of graft survival. In addition, one of seven of the limbal allografts in either group of systemic and topical CsA survived >60 days on cessation of CsA. There was no significant difference in mean survival time between systemic and topical application, although plasma levels of CsA were significantly higher after systemic administration. However, a significant higher aqueous concentration was found in topical treatment. CONCLUSIONS: Limbal allografts were stable in maintaining the reconstructed ocular surface under attentive postoperative immunosuppression. Topically administered CsA was as effective as systemic use.

Scanning electron microscopy of in vitro fertilization in cattle.

Cumulus-oocyte complexes were collected from cows at an abattoir by aspiration from small (1-6 mm) follicles. The complexes were matured in vitro for 28 h. Subsequently, the cumulus cells and the zona pellucida were removed by enzyme treatment in a proportion of the complexes (zona-free ova). Both cumulus-enclosed and zona-free ova were inseminated in vitro and processed for scanning electron microscopy after different periods of culture. In the cumulus-enclosed ova the number of spermatozoa attached to and penetrating into the cumulus investment increased with increasing culture time. Practically all spermatozoa displayed intact acrosomes. In the zona-free ova clusters of spermatozoa attached to the ovum surface, and at 5 h a proportion of the spermatozoa had undergone the acrosome reaction, and their internalization into the ooplasma was initiated. The acrosome reaction was characterized by an increasing fenestration of the membrane coverings of the acrosomal region of the sperm head. During the sperm head internalization, where the ovum microvilli appeared to contact especially the equatorial segment and the postacrosomal region, the sperm head gradually disappeared from the ovum surface, and the microvilli at the site of internalization became more bulbous. Simultaneous abstriction of the second polar body was seen in some ova.

Ultrastructural abnormalities of in vitro fertilization of in vitro matured bovine oocytes.

Cumulus-oocyte complexes collected from cows at an abattoir by aspiration from small (1-6 mm) antral follicles were matured and inseminated in vitro. At different time intervals after insemination the ova were processed for transmission electron microscopy. Up to and including 6 h after insemination all ova were unfertilized, and their cortical granules were more or less clustered. At 6 h acrosome reaction of spermatozoa was observed on the surface of the zona pellucida. At 8 h the first fertilized ovum appeared and the first fully developed spherical pronucleus was observed, at 20 h the first apposition of pronuclei was seen, and at 40 h divisions were ongoing or completed. More than one third of the fertilized ova showed polyspermic penetration of the zona pellucida, and in most of these ova different developmental stages of supernumerary pronucleus formation were observed in the ooplasm. Abnormal cortical granule release was seen in approximately half of the fertilized ova, and it was more frequent in ova with polyspermic as opposed to monospermic penetration of the zona pellucida.

Ultrastructure of the final nuclear maturation of bovine oocytes in vitro.

Cumulus-oocyte complexes were obtained from cow ovaries by aspiration from small (1-6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and subsequently processed for electron microscopy after various periods of culture. By morphological criteria the oocytes could be divided into the following sequence of meiotic stages. The oocyte nucleus I stage was characterized by a spherical nucleus located peripherally in the ooplasm while undulation of the nuclear envelope and initial chromatin condensation was seen at the oocyte nucleus II stage. The oocyte nucleus breakdown stage was characterized by formation of long slender projections from the nuclear envelope in which the envelope doubled back on itself, appearance of dense areas and haphazardly oriented microtubules in the nucleus, marked condensation of the chromatin, and dissolution of the nuclear envelope into irregular vesicles and tubules. The condensed chromatin I stage was characterized by the location of condensed chromatin configuration and uniformly oriented microtubules in a dense area peripherally in the ooplasm while the final condensed chromatin II stage was characterized by a gradual invasion of condensed chromatin configurations into a dense area combined with the presence of the first polar body in the perivitelline space.

Dynamic changes of estradiol and progesterone concentrations during in vitro oocyte maturation in cattle.

Two culture techniques, an open system using 1.0-ml medium and a covered system using 50-mul droplets of medium covered by paraffin oil, used for in vitro bovine follicular oocyte maturation were compared. Estradiol-17beta (E2) was added to Ham's F-10 medium together with other supplementations. In the absence of oocytes, E2 concentration remained unchanged in the 1.0-ml open system, but it decreased gradually in the 50-mul covered system. In the presence of oocytes, E2 increased significantly in the 1.0-ml open system, but this increase could not be detected in the 50-mul covered system. Progesterone (P4) concentration increased in both systems, but it was much higher in the 1.0-ml culture than in the 50-mul culture. The two culture systems allowed an identical nuclear oocyte maturation rate of 88.6 vs 87.0%, a sperm penetration rate of 54.3 vs 59.6%, and a polyspermy rate of 6.8 vs 11.6% for 1.0-ml and 50-mul systems, respectively. The cleavage rate, however, differed significantly (78.3 vs 41.7% for 1.0-ml and 50-mul systems, respectively). It is concluded that diffusion of steroids into the paraffin oil occurs and may affect the cleavage rate but not the maturation or penetration rate.

The sex ratios of bovine embryos produced in vivo and in vitro.

The male:female ratio of developing bovine embryos produced and allowed to develop in vitro and in vivo was determined retrospectively from the cytogenetic analysis of 804 embyos. The overall male:female ratio of the 307 (38%) embryos that could be sexed was 162 (52.8%):145 (47.2%) and did not differ (P>0.05) from the expected 1:1 ratio. Among premorula stage embryos produced in vivo (n = 66) and in vitro (n = 30), the ratios were 1.2:1 and 0.76:1, respectively. Among morulae and blastocysts produced in vivo (n = 74), produced and cultured in vitro (n = 106, and produced in vitro and cultured in vivo (n = 31), the ratios were 1.11:1, 1.3:1 and 0.94:1, respectively, none of which differed significantly from 1:1. There was no difference (P>0.05) in the number of cells or mitotic index between male and female morulae and blastocyst, respectively.

Application of in vitro fertilisation techniques to obtain calves from valuable cows after slaughter.

The ovaries of two infertile cows of high breeding value were recovered after slaughter, and a total of 222 oocytes were obtained. Of these, 156 were classified as of good or fair quality and were subjected to in vitro maturation, in vitro fertilisation (using frozen semen from three bulls of high breeding value) and in vitro culture procedures. After eight days, 27 embryos were obtained, of which 13 were transferred fresh, and 14 were frozen. Three recipients of fresh embryos became pregnant; two calved and one aborted at four months. One of eight recipients of frozen-thawed embryos became pregnant but aborted at three months.

A new flavonol from Sophora tonkinensis.

A new flavonol, tonkinensisol, was isolated from the roots of Sophora tonkinensis, together with three known compounds named as bayin, vitexin and lupeol. Their structures were elucidated on the basis of spectroscopic evidence. Additionally, tonkinensisol showed moderate cytotoxicity suppressing the proliferation of HL-60 cells in vitro.

A detailed analysis of early events during in-vitro fertilization of bovine follicular oocytes.

Bovine follicular oocytes collected at slaughter were matured and fertilized in vitro with in-vitro capacitated spermatozoa. Analysis of 621 penetrated ova fixed at various times after in-vitro insemination led to definition of 6 stages of early development. A time sequence for sperm penetration, sperm head decondensation, male pronucleus formation, the activation of second meiotic division, female chromosome decondensation and pronucleus development was established. First sperm penetration into the ooplasm was recorded 6 h after insemination; 1-2 h was required for the sperm head to decondense and another 4-6 h to develop into the opposing pronucleus stage. Synkaryosis and first cleavage occurred 28 h after fertilization. Examination of the early stages revealed four types of abnormalities, i.e. polyspermy, polygyny, asynchrony between male and female pronucleus development, and preactivation of cytokinesis.

Pregnancy resulting from cattle oocytes matured and fertilized in vitro.

Follicular oocytes (n = 81) collected from cattle at a local slaughterhouse were matured and fertilized in vitro. Of 27 ova 19 (70%) were penetrated by spermatozoa and 40/54 (74%) inseminated ova transferred surgically to the oviducts of a synchronized heifer were recovered by non-surgical flushing of the uterine horns 6 days later. Of the 40 ova 15 (38%) were at the morula, early blastocyst or diminutive morula stages. Culture in vitro sustained further development of all embryos and 9 were expanding or expanded blastocysts. One pregnancy resulted from non-surgical transfer of 2 blastocysts. The results demonstrate that immature oocytes from cattle can be matured and fertilized in vitro, subsequently develop to the blastocyst stage, and develop into a normal pregnancy after non-surgical transfer.