BACKGROUND: As a biochemical marker of cytoplasm, the increased activity of creatine kinase (CK) in human spermatozoa is correlated with both the residual cytoplasma and the ratio of sperm with abnormal func tion. It is a marker of mature sperm and associated with the potential of in semination. OBJECTIVE: To investigate the differences of CK activity in sperm and the relative contents of sperm CK-MM and CK-BB isoenzymes between normal fertile males and infertile males, and evaluate its significance in clinical diagnosis of male infertility. DESIGN: Case-control observation. SETTING: Laboratory of Biochemistry, Family Planning Research Insti tute of Sichuan Province. PARTICIPANTS: Ninety-four male infertile patients between January 1999 and October 2000 were selected from the Department of Family Planning Sciences of Sichuan Province, who had no aspermatism with their wives proved to be fertile. The average age of subjects were 31 years. Eighty subjects with the sperm count ＞ 2×1010 L-1 were taken as normal sperm group and 14 subjects with the sperm counts ＜ 2×1010 L-1 were considered as oligospermia group. Semen obtained from 27 healthy males who were normal in routine examinations and with children was taken as the healthy control group.METHODS: Semen sample collected by masturbation after abstinence of 3 to 5 days was incubated at 37 ℃ for liquefication and routinely analyzed.Total activity of CK in sperm was determined by using a kinetic spectrophotometry and the relative contents of CK isozyme was determined by agarose gel electrophoresis followed by density scanning of CK isozyme.MAIN OUTCOME MEASURES: Sperm counts, percentages of viability and motility of sperm, total CK activity and the relative contents of CK-MM and CK-BB isozyme in spermatozoa.RESULTS: A total of 94 enrolled patients and 27 normal controls were involved in the analysis of results. ①Sperm counts, percentage of viability and motility in oligospermia group ( Ⅱ + Ⅲ, WHO method) were obviously lower than those in the healthy control group, and those in the normal sperm group, except the sperm counts, were remarkably lower than the healthy control group [the sperm counts in healthy control group, normal sperm group and oligospermia group was (6.05 ±0.81 )×1010 L-1, (7.76±1.37)×1010 L-1 and (1.46±0.19)×1010 L-1 respectively (P ＜ 0.01). The survival rate in healthy control group, normal sperm group and oligospermia group was 85.1%,56.8% and 58.2%, P ＜ 0.01, and the sperm motility was 62.9% ,34.6% and 29.5% respectively, P ＜ 0.01].②Total activity of sperm CK in oligospermia group was significantly higher than the healthy control group [respectively (9.000±6.117) and (1.933 ±0.943) kat/108 sperm,P＜ 0.05],although that in the normal sperm group (2.800±0.862) kat/108 sperm was a little higher than the healthy control group, while there were no significant difference between the two groups (P ＞ 0.05).③The relative contents of sperm CK-MM isozyme in the normal sperm group and oligospermia group were obviously decreased (that in the healthy control group, normal sperm group and oligospermia group was 30.5%, 19.0% and 18.0% respectively,P ＜ 0.05), which implied a remarkable difference in sperm differentiation between healthy control group and the latter two groups.CONCLUSION: The determination of sperm CK is meaningful for the diagnosis and pathogenesis of oligospermia. More work should be done on the distribution of sperm CK-MM isozyme in different infertile population as well as its importance in the diagnosis of infertility.

BACKGROUND:Ghrelin is a newly discovered brain-gut peptide from the stomach of human and rats. As an endogenous ligand for growth hormone secretagogue receptor (GHSR), ghrelin can notably stimulate the release of growth hormone. Although GHSR is expressed in many peripheral tissues, little is known about the influence of ghrelin on bone metabolism and GHSR expression in bone tissue. OBJECTIVE:To review the research progress of ghrelin in bone metabolism.METHODS:The first author retrieved CNKI, WanFang, PubMed, and Springerlink databases with the keywords ofGhrelin, bone metabolismin Chinese and English, respectively. The studies regarding ghrelin and its involvement in bone metabolism were included, and repetitive ones were excluded. A total of 53 eligible literatures were selected through skimming abstracts. RESULTS AND CONCLUSION:Ghrelin is a peptide composed of 28 amino acids discovered in gastric endocrine cells and hypothalamic arcuate nucleus in mice and human, which makes a great effect on digestive, nervous, immune and endocrine systems, and also plays a role in hormone secretion, glucose metabolism, immunity, cell proliferation, and inflammation. Serum ghrelin makes a certain influence on bone growth and development, and promotes the differentiation and proliferation of osteoblasts, and inhibits its apoptosis. Additionally, ghrelin suppresses the early osteogenic differentiation of C3H10T1/2 cells by upregulating the expression of Runx2 protein, and attenuates adipogenic differentiation by downregulating PPARγ2 expression, thus inducing osteogenic differentiation. However, few studies have addressed the expression of GHSR in bone tissue.

Objective Co-stimulation cells is a kind of natual killer (NK)-like T cells, which can kill tumor cells.Previous studies show that lupeol , an natural plant extracts , can change the growth of NK cells ,γδT cells and their effects on tumor cells .This study aimed to investigate the effects of lupeol on human co-stimulation cells and colonic cancer cell lines SW 480 . Methods The peripheral blood mononuclear cells ( PBMC) from healthy volunteers were induced in vitro by different cytokines and transferred into Co-simulation cells.After SW480 and Co-stimulation cells were incubated with different concentrations of lupeol for different durations , methyl thiazolyl tetrazolium (MTT) was used to detect the effects of lupeol on co-stimulation cells and colonic cancer cell lines SW480. Lactate dehydrogenase was used to determine the cell-killing activity of Co-simulation cells to colonic cancer lines SW 480 . Results The concentration of lupeol in 0.1-200.0 mg/L promoted the growth of Co-stimulation cells and inhibited the colonic cancer cell lines SW480.When the concentration of lupeol is at 12.5 mg/L, the cell-killing activity of Co-simulation cells to colonic cancer lines SW 480 was enhanced significantly compared with the controls (76%vs 40%, P<0.05). Conclusion Lupeol could promote the prolifera-tion of Co-stimulation cells, inhibit the growth of cancer lines SW480, and strengthen the cytotoxicity of Co-stimulation cells against co-lonic cell lines SW480.

Objective To establish a PCR-RFLP method for detection of CD26 mutation of hemoglobin E(HbE)gene.Methods The genome DNA from the family members who were suspected of having HbE combined with beta-thalassemia was amplified using PCR.The PCR products were digested by restriction endonuclease Mnl I,and then separated on PAGE.The electrophoretical patterns were finally analyzed to confirm if CD26 mutation was present or not.Results Four cases with CD26 mutation in two families were successfully screened out.Conclusion Although the incident frequency of CD26 mutation in Sichuan population is not high,it must not be neglected since it may present alone or combined with other type of thalassemia.PCR-RFLP method described in this study is available in screening mutated HbE gene and treating the disease owing to its simplicity,rapidity and specificity.

Objective To explore a more efficient surgical method to cure axillary osmidrosis through a comparative study on results from two surgical treatments of axillary osmidrosis in the armpit area.Methods 72 patients were randomized into two groups,in which 36 patients (group A) with micro-incision simple liposuction and 36 patients (group B) with dissection and excision of subcutaneous tissue from axillary folds.The clinical outcomes in the two groups were comparatively analyzed,based on surgery time,effect,complications,scarring and recovery time.Results The total efficiency of axillary osmidrosis treatment was 100％ after 6-12 months clinical follow-up towards two groups.There was no significantly statistical difference on the effect of both groups.Micro-incision simple liposuction was obviously superior to dissection and excision of subcutaneous tissue from axillary folds based on surgery time,complications,recovery time,scarring and armpit hair survival rate.The two groups were significantly different (P＜0.05).Conclusions Minimally invasive simple liposuction,as a treatment for axillary osmidrosis is a method with more effectiveness and reliability.It is a more acceptable approach to patients in view of safety and convenience,smaller and invisible scarring,fewer complications,and less recovery time.

AIM: To study the role of endothelin-1 (ET-1) in portal hypertension (PHT) induced by endotoxin. METHODS: Collagenase in situ perfusion was adopted to separate hepatic stellate cells (HSCs). HSCs was cultured on concretized collagen. ET-1 anti-sense oligonucleotide was added into the culture medium and then LPS was also added up to the concentration of 1 000 ?g/L. The diameters of the concretized collagen were measured. Sense and mis-sense oligonucleotide were applied as control. ET-1 in the culture medium was detected by radioimmunoassay and ET-1 mRNA in HSCs was detected by RT-PCR. ?-actin of HSCs was detected by Western blotting. RESULTS: The diameter of concretized collagen on which HSCs pretreated with ET-1 anti-sense oligonucleotide was 93.3%?3.8% the size of the primary. The diameter of concretized collagen of the control groups were 70.1%?4.8% and 70.5%?3.9% (P

This study aims to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 using the orthogonal design method, and to investigate its effects of the component formula on cell proliferation, apoptosis and cytoskeleton in lung cancer A549 cells. The orthogonal design method was introduced to optimize the most effective component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on lung cancer A549 cells. CCK-8 assay and Real-time cell analysis were adapted to analyze the effect of component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma on A549 cells viability at different time and dose. Cell apoptosis was measured by Annexin V- FITC/PI double staining and flow cytometry. Cell skeleton protein F-actin was detected by high content screening (HCS). The optimizing component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma for total salvianolic acid, total saponins of panax ginseng and ginseng polysaccharide doses were 5, 10, 5 mg L(-1). CCK-8 assay and real-time cell analysis demonstrated that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma treatment could significantly decrease the A549 cell viability in both dose- and time-dependent manner compared with control group (P < 0.01). Moreover, the increase of cell apoptosis was detected by Annexin V-FITC/PI double staining and flow cytometry when cells treated with the component formula, which indicating that the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma could induce A549 cell apoptosis in a time-dependent manner compared with control group (P < 0.01). Furthermore, compared with control group, a significant decrease in A549 cell skeleton area was found in the component formula-exposed cells in the dose-dependent manner (P < 0.01). In summary, the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma inhibits A549 cell proliferation by inducing cell apoptosis and decreasing cell microfilament formation. All of these results will be helpful to reveal antitumor mechanism of the component formula of Salviae Miltiorrhizae Radix et Rhizoma and Ginseng Radix et Rhizoma, which provides a basis for the exploration of antitumor mechanism of the component formula on lung cancer.
Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; Panax ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; Plant Roots ; chemistry ; Rhizome ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective To compare and analyze the clinical characteristics of acute central cervical spinal cord injury with only upper extremity involvement and with both upper and lower extremity involvement.Methods A retrospective case control study was made on clinical data of 76 patients with acute central cervical spinal cord injury hospitalized from January 2010 to December 2013.Nerve injury involved was only upper extremity in 39 patients (upper extremity group),but both upper and lower extremities in 37 patients (upper-and lower-extremity group).In upper extremity group,there were 35 males and four females,age was 21-80 years [(52.5 ± 13.4) years],injury resulted from traffic accidents in 24 patients,ground-level falls in eight,high-level falls in six and heavy-object hit in one,and level of injury was C3/4 in 16 patients,C4/5 in 14 and C5/6 in nine.In upper-and lower-extremity group,there were 30 males and seven females,age was 36-78 years [(59.6 ± 9.7) years],injury resulted from traffic accidents in 16 patients,ground-level falls in 11,high-level falls in seven and heavyobject hit in three,and level of injury was C3/4in nine patients,C4/sin 18 and C5/6in 10.Sagittal diameter of the cervical spinal canal,maximal canal compromise,maximal spinal cord compression,degenerating factors of the cervical spine and treatment protocols were determined.Upper extremity function was assessed with the American spinal injury association (ASIA) score.Results There were significant differences between upper extremity group and upper-and lower-extremity group in sagittal diameter of the cervical spinal canal [(7.5 ± 1.5)mm ∶ (6.8 ± 1.2)mm],maximal canal compromise [(28.9 ±9.6)％ ∶ (34.9 ± 10.6)％],ASIA score at admission[(31.6 ± 11.8)points ∶ (22.7± 11.3)points)] and ASIA score at last follow-up [(46.2 ± 4.2) points ∶ (40.2 ± 4.0) points] (P ＜ 0.05),while the maximal spinal cord compression in upper extremity group [(15.7 ± 11.9)％] had no significant difference from that in upper-and lowerextremity group [(17.0 ± 10.6) ％] (P ＞ 0.05).Lower prevalence of posterior osteophyte of the vertebral body was noted in upper extremity group than upper-and lower-extremity group (15％ ∶ 51％) (P ＜0.01).Twenty patients (49％) in upper extremity group were surgically treated,while 31 patients (84％) in upperand lower-extremity group (P ＜ 0.05).Conclusions Compared to acute central cervical spinal cord injury with both upper and lower extremity involvement,the injury with only upper extremity involvement is much common in younger patients and is characterized by lowered frequency of osteophyte,large buffer space,mild nerve damage,preferred non-operation treatment and good prognosis.

Aim To optimize the most effective compo-nent formula from the active ingredients of Salvia Milti-orrhiza and Panax Ginseng through the orthogonal de-sign method to resist breast cancer, and to reveal its antitumor mechanism in MCF-7 cells. Methods The human breast cancer cells MCF-7 were employed as the research object and the normal breast epithelial cells MCF-10A were used as control,optimizing the most ef-fective component formula from the active ingredients of Salvia Miltiorrhiza and Panax Ginseng by using CCK-8 assay and orthogonal design method; real-time cell a-nalysis was used to monitor the best combination formu-la on cell proliferation, and high content screening was used to detect the best combination drug on cell apop-tosis. Results The best combination of the salvianolic acids, saponins of Panax Ginseng and ginseng polysac-charides that were screened out were 5 , 10 , 5 mg · L-1 . Compared with control group, the treatment group had effective response inhibiting the proliferation on MCF-7 cells, but those effects were weaker on MCF-10A cells through real-time cell analysis. Ho-echst, Annexin V, PI staining fluorescence showed no significant difference ( P >0. 05 ) on MCF-10 A cells compared with the control group,but there was signifi-cant difference ( P <0. 01 ) on MCF-7 cells by HCS. Conclusions The most effective component formula from the active ingredients of Salvia Miltiorrhiza and Panax Ginseng have a strong inhibition of proliferation and induction of apoptosis to resist breast cancer with selection, and there is no significant difference in MCF-10A cells.

Objective:To explore the distribution characteristics of respiratory pathogens in patients with community-acquired pneumonia in Lianyungang.Methods:A total of 612 patients admitted to the second people′s Hospital of Lianyungang City because of community-acquired pneumonia (CAP) in 2019 were selected as subjects. Sputum or pharyngeal swabs were collected to extract nucleic acids, and 13-fold nucleic acids of respiratory pathogens were detected by PCR capillary electrophoresis fragment analysis. SPSS statistical software and GraphPad5.0 statistical mapping software were used for statistical analysis.Results:The physical examination rate of respiratory pathogens in the adult group was 82.0% in winter, 48.4% in spring, 28.0% in autumn, 20.0% in summer, χ 2=38.473, P=0.000. The positive rate of nucleic acid detection was significantly different in different seasons, among which the physical examination rate of respiratory pathogens in winter was the highest. The physical examination rate of respiratory pathogens in the juvenile group was 86.0% in spring, 76.2% in winter, 71.3% in summer and 66.7% in autumn, χ 2=7.946, P=0.047 . The positive rate of nucleic acid detection was calculated according to gender grouping. The comparison of nucleic acid positive rate between adult group and juvenile group in different seasons: 86.0% vs 48.4% in spring, χ 2=19.436, P=0.000; 71.3% vs 20.0% in summer, χ 2=22.180, P=0.000; 66.7% vs 28.0% in autumn, χ 2=13.485, P=0.000; 76.2% vs 82.0% in winter, χ 2=0.758, P=0.384. Except in winter, the detection rate of nucleic acid of pathogens in the juvenile group was significantly higher than that in the adult group. Conclusions:The nucleic acid detection rate and etiological distribution characteristics of respiratory pathogens are different in patients with community-acquired pneumonia in different seasons and different age groups. 13 kinds of multiple detection methods of respiratory pathogens can provide favorable laboratory data support for the diagnosis and treatment of clinical CAP patients.