Objective To examine neuroinflammation,oxidative damage and neuron loss in the contralateral parie-tal lobecortex of TBI model mice, and to investigate effects of berberine chloride on such secondary damage.Methods TBI model was established by a weight-drop hitting device and mice in berberine group were administered intragastrically with berberine chloride (50mg/kg.day) for 21 days.Immunofluorescence staining was used to assess activity of microglia and astrocyte.Immunohistochemistry was used to assess DNA oxidative damage, neuron loss and expression of COX-2 and iN-OS.Results Activation of microglia and astrocyte, expressions of COX-2 and iNOS and DNA oxidative damage were ob-viously increased by TBI,(19.82 ±1.88)and(16.96 ±1.69)、(13.79 ±4.32)and(8.67 ±0.96)、(27.86 ±5.38) and (16.00 ±7.59)、(31.92 ±6.57)and(24.79 ±2.78)respectively (P<0.01 or P<0.05).Activation of microglia and ex-pressions of COX-2 and iNOS were significantly suppressed by berberine ,(15.49 ±1.88)and(19.82 ±1.88)、(16.83 ± 7.89)and(27.86 ±5.38)、(26.25 ±2.41)and(31.92 ±6.57) respectively(P<0.01 or P<0.05).There was no differ-ence in neuron loss among three groups, (49.05 ±4.38),(48.56 ±3.56)and (47.75 ±4.14) respectively (P>0.05). Conclusions TBI can cause neuroinflammation and oxidative damage but not neuron loss in the contralateral parietal lobe cortex.Berberine chloride can significantly suppress neuroinflammtion in the contralateral parietal lobe cortex after TBI.

OBJECTIVESTo evaluate the clinical significance of the detection of IgG and IgM antibodies against Chlamydia trachomatis (CT) in semen of asymptomatic infertile patients.

METHODSOne hundred and sixteen asymptomatic infertile patients and eighteen fertile males were selected randomly. The routine parameter analysis of semen was fulfilled by computer aided semen analysis(CASA). Then the seminal plasma was separated and the IgG, IgM antibodies against CT in seminal plasma were determined with ELISA method.

RESULTSIgG and IgM antibodies against CT were present in 13.8% (16/116) and 3.4% (4/116) of the semen of infertile patients, while for the fertile males the percentages were 11.1% (2/18) and 0, respectively. There were no differences between the two groups(P > 0.05). In the infertile patients, 22 patients were azoospermia. And in the rest 94 infertile patients, the percentages of IgG and IgM antibodies in abnormal sperm density group were 21.4% (6/28) and 7.1% (2/28), which were higher than those in normal group, but there were no statistical differences(P > 0.05). Similarly, the IgG, IgM antibodies were not correlated with the sperm motility(P > 0.05). The positive percentage of CT in 116 patients was 25.9% (30/116).

CONCLUSIONSThe percentages of IgG and IgM antibodies against CT in semen of asymptomatic infertile patients are similar to that in fertile males, which do not correlate with the changes of semen parameters, and may not be used for indication of CT infection.

Adult ; Antibodies, Bacterial ; blood ; Chlamydia trachomatis ; immunology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Infertility, Male ; microbiology ; Male ; Sperm Count ; Sperm Motility

Animals ; Biological Warfare ; Burns ; pathology ; Burns, Chemical ; pathology ; Chemical Warfare ; China ; Humans ; Military Medicine ; Radiation Injuries ; pathology ; Wounds, Gunshot ; pathology

Men with non-obstructive azoospermia(NOA) can now be treated by using intra-oocyte round spermatid injection(ROSI) or elongated spermatid injection(ELSI). Spermatids can be retrieved from semen or from testis biopsy specimens. But the rates of fertilization and pregnancy with spermatids have been disappointing. Many problems limiting success rate and hindering a wide application of this technique still remain unresolved, including the incomplete maturation of spermatid nuclear, oocyte activation and identification of a live spermatid.
Azoospermia ; therapy ; Cryopreservation ; Female ; Humans ; Male ; Pregnancy ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; cytology

AIMTo analyze the relationship between autosomal aberrations and testicular dysgenesis or spermatogenic arrest in Chinese patients and to map the corresponding regions on each autosome in regard to the recorded aberrations accompanying these distubances.

METHODSOne hundred and nineteen cases of aberrant karyotypes with testicular dysgenesis, azoospermia or oligozoospermia reported in five Chinese journals and one monograph were analyzed. For each autosome, the type and frequency of chromosomal aberrations were counted and the regions corresponding to the disturbances were mapped out.

RESULTSChromosomes 13, 14, 9, 21 exhibited a high frequency of aberration and bands 14q11 and 13p11 were the two regions showing the highest linkage to testicular dysgenesis or infertility. The frequency of chromosomal aberrations was higher in bands 9p11 and 22q than in others.

CONCLUSIONAutosomes 13, 14, 9 and 21 in the order of importance play a critical role in testicular development and spermatogenesis and other autosomes may also contribute; the following regions, 14q11, 13p11,9p11, and 22q, are of high significance.

Asian Continental Ancestry Group ; Chromosome Aberrations ; Chromosomes, Human, Pair 13 ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 9 ; Gonadal Dysgenesis ; genetics ; Humans ; Infertility, Male ; genetics ; Karyotyping ; Male ; Oligospermia ; genetics ; Testis ; abnormalities

OBJECTIVESTo develop a simple and effective method by which spermatids can be isolated from mouse testis.

METHODSCombination of enzymatic digestion was used to prepare suspension of spermatogenic cells from adult mouse testis, and then a modified discontinuous Percoll gradient (15%, 22%, 30%, 40%, 50%, 60%) centrifugation method was introduced to isolate spermatids from the cellular suspension. The content of spermatids in each isolated fraction by Percoll method was determined by morphology (Wright-Giemsa staining) and flow cytometry analysis, and the viability of spermatogenic cells was assessed using Eosin Y exclusion test.

RESULTSMore than 97% of the testicular cells remained their viability after enzymatic digestion. After Percoll centrifuged, six fractions were formed. In each isolated fraction, the 22% fraction contained mostly spermatids(mean 86.7%) and cell viability was more than 85.5%. While in the 30% fraction, immature spermatogenic cells were present, and more than 92% of the cells remained their viability.

CONCLUSIONSA large of relatively purified spermatids can be isolated from mouse testis by enzymatic digestion combined discontinuous Percoll gradient centrifugation method.

Animals ; Cell Separation ; methods ; Centrifugation, Density Gradient ; methods ; Male ; Mice ; Spermatids ; cytology ; Testis ; cytology

Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.

OBJECTIVETo develop a nested polymerase chain reaction (PCR) technique for fetal SRY gene identification using cell-free fetal DNA in maternal plasma.

METHODSPeripheral blood samples were obtained from 30 pregnant women and cell-free DNA was extracted by the phenol/chloroform method from plasma. The nested PCR was carried out to amplify the fragment of SRY gene by two sets of PCR primer pairs. Direct sequencing analysis was then performed on the PCR product.

RESULTSAmong the 17 women bearing male fetuses, SRY sequences were detected in 15 plasma samples after nested PCR amplification, while none of the 13 women bearing female fetuses had the positive results. The accuracy and sensitivity were 93.3% (28/30) and 88.2% (15/17), respectively.

CONCLUSIONThe phenol/chloroform extraction for fetal DNA in maternal plasma was effective and simple. And the nested PCR amplification of SRY sequence is a convenient and low-cost approach for the non-invasive early prenatal diagnosis of sex-linked inheritant diseases.

Adult ; Base Sequence ; DNA ; blood ; genetics ; Female ; Fetus ; Genes, sry ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Pregnancy ; blood ; genetics ; Prenatal Diagnosis ; Sensitivity and Specificity

OBJECTIVETo investigate the antitumor efficacy of death receptor 5, its ligand (TRAIL) and DR5mAb in human hepatocellular carcinoma.

METHODSExpression of DR5 in the HCC cell lines HepG2, SMMC 7721 and normal human liver cell line LO2 was measured at mRNA and protein level by semi-quantitative RT-PCR and Western blot, respectively. MTT method was used to measure the cell viability and flow cytometry assay was used to detect apoptosis so as to observe the inhibitory effect of TRAIL and DR5mAb on HCC cells.

RESULTSDeath receptor 5 was highly expressed in the HCC cell lines, but rarely expressed in normal human liver cell line (P < 0.01). With the increase of TRAIL concentration, the cell viability of HCC cells decreased gradually. However, when the concentration of TRAIL was above 1000 ng/ml, HCC cells were resistant to TRAIL, but still sensitive to DR5mAb. After incubation with DR5mAb (1000 ng/ml) for 24 h, the rate of apoptosis in HCC cells reached to 52.45% +/- 0.57%, which was higher than that incubated with TRAIL under the same condition (14.74% +/- 0.48%) (P < 0.05). The cell viability of normal human liver cell line treated with TRAIL tended to decline with the increase of the concentration, which was significantly different from that of matched control group. But DR5mAb had little effect on normal human liver cell line.

CONCLUSIONDeath receptor 5 as a target plays an important role in the course of HCC apoptosis induction. Agonistic monoclonal antibody specific for human DR5 can selectively and effectively kill hepatocellular carcinoma cells in vitro, while is not harmful to normal human hepatocytes. It reveals that DR5mAb might provide a new direction in hepatocellular carcinoma treatment research.

Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line ; Cell Line, Tumor ; Cell Survival ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; genetics ; immunology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis

OBJECTIVETo determine the expression of nerve growth factor (NGF) in hepatic tissues and serum of patients with primary liver cancer (PLC), and to investigate the relationship of serum NGF levels with clinicopathological features of PLC.

METHODSHepatocellular carcinoma (HCC) samples and patient-matched tumor-adjacent liver samples were collected from 26 PLC patients to assess the mRNA and protein expressions of NGF by reverse transcription-PCR, western blotting (b-actin normalized), and immunohistochemistry. In addition, serum samples were collected from 40 PLC patients, 40 liver cirrhosis patients, 40 chronic hepatitis patients (including hepatitis B or C virus infections), and 30 healthy (normal) controls. The serum levels of NGF were measured by enzyme-linked immunosorbent assay. Intergroup differences were assessed by the t-test, and correlation with sex, age, presence of cirrhosis, tumor size, and TNM classification were assessed by the Kruskal-Wallis H test followed the Mann-Whitney U test.

RESULTSHCC tissues showed higher mRNA and protein expressions of NGF than the corresponding tumor-adjacent non-HCC tissues. Hepatic NGF expression was mainly localized to the tumor cell cytoplasm. Serum NGF expression was significantly higher in PLC patients (33.86+/-16.11 pg/ml) and cirrhosis patients (20.57+/-9.73 pg/ml) than in normal controls (11.13+/-6.12 pg/ml) and chronic hepatitis patients (13.20+/-6.23 pg/ml) (P less than 0.01). Furthermore, when the PLC patients were stratified according to tumor size and TNM stage, the serum NGF level was found to be significantly higher in patients with tumors more than 5 cm (vs. less than 5 cm; U=83.000, P=0.002) or of TNM stage III/IV (vs. stage I/II; U=103.500, P=0.009).

CONCLUSIONElevated expression of NGF in liver cancer tissues and serum of PLC patients is related with tumor size and TNM staging. These findings suggest that NGF may play a role in HCC tumorigenesis and/or that serum NGF may represent a prognostic marker of PLC.

Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Case-Control Studies ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Nerve Growth Factor ; blood ; metabolism ; Prognosis ; Young Adult