BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26％ of cells were positive for CD29,7. 51％ for CD34 and 4. 02％ for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.

OBJECTIVETo observe the clinical efficiency of curettage combining circumcision on giant condyloma acuminata(CA) in male external genitalia, the relationship between recurrence and curettage depth, the possibility of HPV infection in PBMC after operation.

METHODSCurettage combining circumcision was carried out on 50 cases with CA. The removed wart and wound surface tissues were examined under light microcope and for HPV-DNA detection by PCR. HPV-DNA was detected in PBMC during < or =1 and > or =2 weeks after operation.

RESULTS(1) 46 cases were cured completely after one treating (once the cure rate is 92%), 4 cases twice(twice the cure rate is 8%); (2) The tissues were proved to be HPV-DNA positive by PCR amplification and CA relapse occurred; (3) HPV-DNA was detected in PBMC only within the week after curettage in the 19 cases.

CONCLUSIONS(1) The clinical efficiency of curettage combining circumcision on giant condyloma acuminata in male external genitalia is sure; (2) Cure rate and relapse rate are related with curettage depth; (3) Transient positive HPV-DNA in PBMC may be detected.

Adolescent ; Adult ; Aged ; Circumcision, Male ; Condylomata Acuminata ; surgery ; Curettage ; methods ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo observe the interventional effect of electroacupuncture combined with catgut implantation at acupoints for treatment of simple obesity of heart and spleen deficiency type.

METHODSSixty five cases were randomly divided into an observation group (33 cases) and a control group (32 cases). The observation group was treated with electroacupuncture combined with catgut implantation at acupoint therapy, the electroacupuncture was applied at Zhongwan (CV 12), Xiawan (CV 10), Guanyuan (CV 4), Tianshu (ST 25), ect. and catgut implantation was given at Zhongwan (CV 12), Tianshu (ST 25), Qihai (CV 6), etc. The control group was treated with electroacupuncture only. The body weight, body mass index (BMI), waistline, waist hip ratio (WHR), Pittsburgh sleep quality index (PSQI), Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD, 17 items) were evaluated before and after treatment, and these were also compared with those of 35 nomal cases.

RESULTSThe total effective rate of 93.9% in the observation group was higher than that of 84.4% in the control group (P < 0.05); the body weight, BMI, waistline, WHR, PSQI, HAMD and HAMA of simple obesity cases were obviously higher than those of normal cases (all P < 0.05). The scores of above indexes were all obviously decreased in both groups after treatment (all P < 0.05), and the improvement was more significant in observation group (P < 0.05).

CONCLUSIONThe sleep quality reduction and mental and psychology disorder exist in simple obesity patients, and electroacupuncture combined with catgut implantation at acupoints can reduce weight effectively, and at the same time improve the sleep quality and regulate psychological state.

Acupuncture Points ; Adult ; Catgut ; Electroacupuncture ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Obesity ; physiopathology ; psychology ; surgery ; therapy ; Prostheses and Implants ; Prosthesis Implantation ; Spleen ; physiopathology ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the effect of transplantation of allogeneic adipose-derived stem cells (ADSCs) on serum biochemical indicators and sporting ability in highly endurance-trained rats from a fasciological perspective.

METHODSThe ADSCs were cultured in vitro. Forty male Wistar rats were randomized into 4 equal groups, namely the blank control group, overtraining (model) group, transplantation without training group and overtraining plus transplantation group. The rats in the two overtraining groups were subjected to exhaustive swimming for 1 week, and in the two transplantation groups, cultured allogeneic ADSCs (2×10(6)/ml) were injected via the tail vein. The exhaustion time in swimming and the serum levels of BUN, LDH, BLa, and Hb of the rats were recorded after the treatments.

RESULTSThe rats in the model group showed significantly increased serum BUN, LDH and BLa levels and decreased Hb level with a extended exhaustion time as compared with those in the blank control group (P<0.01). The BUN, LDH and BLa was significantly lower, Hb level higher and the exhaustion time significantly longer in the overtraining plus transplantation group than those in the model group (P<0.01).

CONCLUSIONSADSCs can effectively prolong the exhaustion time of rats during exhaustive swimming and enhance their sporting ability.

Adipose Tissue ; cytology ; Animals ; Blood Urea Nitrogen ; Fatigue ; prevention & control ; Hydro-Lyases ; blood ; Lactic Acid ; blood ; Male ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Swimming

OBJECTIVETo investigate the effects of systemic transplantation of allogenic adipose-derived stem cells (ADSCs), the main undifferentiated cells in the supporting-storing system based on the fasciology hypothesis, on the bone mineral density (BMD) and histomorphometry in rats with glucocorticoid-induced osteoporosis (GIOP) rats, and explore a new therapeutic approach for osteoporosis.

METHODSForty female adult Wistar rats were randomized equally into blank control group (A), model group (B), control treatment group (C) and treatment group (D). In groups B, C, and D, osteoporosis was induced by injection of prednisolone (8 mg/kg) via the tail vein 3 times a week for 12 consecutive weeks. After successful establishment of osteoporosis, allogenic ADSCs (3×10(6)/ml) were transplanted via the tail vein. The BMD at the L3-L5 levels and of the right femurs were detected, and the histomorphometry of the right tibias was analyzed in all the rats.

RESULTSAfter prednisolone injection, the BMD of L3-L5 vertebrae and the right femurs, the percent trabecular area, trabecular thickness and trabecular number of the right tibias were all obviously lowered while the trabecular separation and osteoclast number increased in group B as compared to those in group A (P<0.05). Four weeks after ADSC transplantation, all these indices were significantly improved in group D, showing significant differences from those in group B (P<0.05) and also group C.

CONCLUSIONTransplantation of allogenic ADSCs can restore the BMD and bone histomorphometric properties of rats with GIOP, and may serve as a potential treatment for GIOP. These results also provide partial experimental evidence supporting the hypothesis of fasciaology.

Adipocytes ; cytology ; Animals ; Bone Density ; Female ; Glucocorticoids ; adverse effects ; Osteoporosis ; chemically induced ; surgery ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; Transplantation, Homologous

Two plasmid constructs, pcE2 and pcE3, containing 3' fragment of open reading frame 2 (ORF2,1163 bp) of hepatitis E virus (HEV) and full-length ORF3 (369 bp), were injected into bilateral tibialis of Swiss mice respectively，for three times (0, 2nd and 4th weeks) and observed the HEV IgG by ELISA. HEV IgG was induced after the injection of pcE2 or pcE3 or both, and the percentage of seraconversion was 100% after two weeks of the third injection. Compared with injection of either construct, the antibody titers were higher in the group with combined injection of two constructs.

OBJECTIVETo investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.

METHODSSW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.

RESULTSCompared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).

CONCLUSIONRadiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; pathology ; Humans ; Kisspeptins ; genetics ; metabolism ; radiation effects ; RNA, Messenger ; genetics ; X-Rays

OBJECTIVE: To determine the distribution of pathogens and their characteristics of drug susceptibility originating from nosocomial infections in the intensive care units (ICU), and to provide evidence for clinical anti-infection treatments. METHODS: Retrospective analysis to the pathogens and their drug susceptibility characteristics was carried out. These pathogens were isolated from the samples that came from patients infected in the ICU from 2002 to 2004. RESULTS: The main nosocomial infective pathogens in the ICU were gram negative bacilli (48.2%), and the next ones were gram positive bacteria (43.3%) and fungus (8.5%). The most common gram negative bacilli were Pseudomonas aeruginosa and Escherichia coli; while for gram positive bacteria, the main bacterin were Staphylococcus aureus. The gram negative bacilli could resist 4 or more than 4 antibiotics, and the rate for resistance exceeded 40%. Similarly, oxacillin resistance staphylococcus could resist 7 antibiotics, and the rate was over 50%. The detective rates of ESBLs and AmpC enzymes produced by Escherichia coli and K. peumoniae were 34.0% & 30.7% and 13.2% & 23.1%, respectively. The rate for oxacillin resistance staphylococcus was 66.3%, and there was relative high resistance rate ( > 55%) for most antibiotics: There was statistical difference, compared with that of non-resistant strains. CONCLUSION Though gram positive coccus still play an important role, most infections are caused by gram negative bacilli of nosocomial infections in the ICU. The antibiotics resistant rate of all bacteria has been rising gradually. It shows strong resistance and multi-drug resistance. The most importment cause for resistance of gram negative bacilli is that the bacteria can produce ESBLs and AmpC enzymes. The antibiotic resistant rate for oxacillin resistance staphylococcus is really high.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Cross Infection ; microbiology ; Drug Resistance, Microbial ; Drug Resistance, Multiple ; Escherichia coli ; drug effects ; isolation & purification ; Female ; Gram-Negative Bacteria ; drug effects ; isolation & purification ; Humans ; Intensive Care Units ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Oxacillin ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; isolation & purification ; Retrospective Studies ; Staphylococcus aureus ; drug effects ; isolation & purification

OBJECTIVETo observe the clinical effect of dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises for treatment of female stress urinary incontinence.

METHODSSeventy one cases were randomly divided into two groups. Thirty six cases in the observation group were treated with acupuncture on Zhongji (CV 3), Zigong (EX-CA 1), Chize (LU 5) etc. and tortoise-shell moxibustion on Shenque (CV 8) combined with pelvic floor muscle exercises; while thirty five cases in the control group were treated with only pelvic floor muscle exercises. The scores of the International Consultation Committee on Incontinence Questionnaire Short Form (ICI-Q-SF) and the Medical Outcomes Survey Short Form-36 (SF-36) were evaluated before and after treatment, and the scores of SF-36 were also compared with 35 cases in normal group.

RESULTSThe total effective rate of 91.7% in the observation group was higher than that of 77.1% in the control group (P < 0.05). The dimensions of SF-36 of stress urinary incontinence patients were remarkably lower than those of normal group (all P < 0.05). The scores of ICI-Q-SF were decreased while the scores of SF-36 were increased obviously after treatment in both the observation group and the control group, there were pronounced improvements on physiological function, pain, physical activity, social function and affection function in the observation group (all P < 0.05).

CONCLUSIONThe quality of life for female stress urinary incontinence patients may be poor, however the dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises can improve the symptoms of urinary incontinence and increase the quality of life of patients.

Acupuncture Therapy ; Adult ; Aged ; Exercise Therapy ; Female ; Humans ; Middle Aged ; Moxibustion ; Muscle Contraction ; Muscles ; physiopathology ; Treatment Outcome ; Urinary Incontinence, Stress ; physiopathology ; therapy

OBJECTIVETo observe the influence of auricular point sticking on incidence of nausea and vomiting and analgesia effect after gynecological laparoscopy, and provide evidence for clinical application of auricular point sticking.

METHODSOne hundred and twenty cases of selective gynecological laparoscopy under general anesthesia were randomly divided into an auricular point sticking group and a placebo group, 60 cases in each group. In the auricular point sticking group, the auricular point sticking with vaccaria seeds was applied at Shenmen (TF 4), Wei (CO 4) and Jiaogan (AH 6a) before the operation and 1, 5, 9, 23 h after the operation, which were pressed 5 min each point each time. The two ears were proceeded at the same time. In the placebo group, the same point selection, sticking paste was used as the auricular point sticking group, but no sticking or pressing with vaccaria seeds was adopted. The incidence of nausea and vomiting, the usage rate of tropisetron and morphine within 24 hours of the operation, as well as the score of visual analogue scale (VAS) and other adverse reactions at 2, 6, 10, 24 h after the operation were observed respectively.

RESULTSCompared with the placebo group, the incidence of nausea and vomiting [31.7% (19/60), 16.7% (10/60) vs 58.3% (35/60), 35.0% (21/60)], the usage rate of tropisetron [21.7% (13/60) vs 48.3% (29/60)] and morphine [18.3% (11/60) vs 38.3% (23/60)], the VAS scores at all different time points in the auricular point sticking group were all decreased (all P < 0.05), and no adverse reaction was observed.

CONCLUSIONThe auricular point sticking could significantly decrease the incidence of nausea and vomiting in patients of gynecological laparoscopy and has positive analgesic effect.

Acupuncture Analgesia ; Acupuncture Points ; Acupuncture, Ear ; Adult ; Female ; Genital Diseases, Female ; surgery ; Gynecology ; Humans ; Laparoscopy ; adverse effects ; Middle Aged ; Nausea ; etiology ; therapy ; Vomiting ; etiology ; therapy ; Young Adult