Modeling Syphilis and HIV Coinfection: A Case Study in the USA.

Syphilis and HIV infections form a dangerous combination. In this paper, we propose an epidemic model of HIV-syphilis coinfection. The model always has a unique disease-free equilibrium, which is stable when both reproduction numbers of syphilis and HIV are less than 1. If the reproduction number of syphilis (HIV) is greater than 1, there exists a unique boundary equilibrium of syphilis (HIV), which is locally stable if the invasion number of HIV (syphilis) is less than 1. Coexistence equilibrium exists and is stable when all reproduction numbers and invasion numbers are greater than 1. Using data of syphilis cases and HIV cases from the US, we estimated that both reproduction numbers for syphilis and HIV are slightly greater than 1, and the boundary equilibrium of syphilis is stable. In addition, we observed competition between the two diseases. Treatment for primary syphilis is more important in mitigating the transmission of syphilis. However, it might lead to increase of HIV cases. The results derived here could be adapted to other multi-disease scenarios in other regions.

Modeling and Research on an Immuno-Epidemiological Coupled System with Coinfection.

In this paper, a two-strain model with coinfection that links immunological and epidemiological dynamics across scales is formulated. On the with-in host scale, the two strains eliminate each other with the strain having the larger immunological reproduction number persisting. However, on the population scale coinfection is a common occurrence. Individuals infected with strain one can become coinfected with strain two and similarly for individuals originally infected with strain two. The immunological reproduction numbers [Formula: see text], the epidemiological reproduction numbers [Formula: see text] and invasion reproduction numbers [Formula: see text] are computed. Besides the disease-free equilibrium, there are strain one and strain two dominance equilibria. The disease-free equilibrium is locally asymptotically stable when the epidemiological reproduction numbers [Formula: see text] are smaller than one. In addition, each strain dominance equilibrium is locally asymptotically stable if the corresponding epidemiological reproduction number is larger than one and the invasion reproduction number of the other strain is smaller than one. The coexistence equilibrium exists when all the reproduction numbers are greater than one. Simulations suggest that when both invasion reproduction numbers are smaller than one, bistability occurs with one of the strains persisting or the other, depending on initial conditions.

[Effects of Electroacupuncture and Intracerebral Injection of VEGF on Caspase12, Caspase3, and GRP78 Genes in Rats with Cerebral Ischemia-Reperfusion Injury].

OBJECTIVE: To determine the effects of electroacupuncture (EA) and intracerebral injection of vascular endothelial growth factor (VEGF) on caspase12, caspase3, and glucose regulated protein 78 kD (GRP78) genes of rats with cerebral ischemia reperfusion injury. METHODS: 60 SD rats were randomly divided into sham-operation group, model group, EA group and EA+VEGF group with 15 rats in each group. Middle cerebral artery occlusion (MCAO) method was used to establish the model of cerebral ischemia reperfusion injury. Electro-acupuncture intervention was introduced 1 day after the injury in the EA group and EA+VEGF group: 30 minutes each session and once a day for a total of 14 d [acupoint selection: Baihui (GV 20), Quchi(Li 11), Zusanli (ST36)]. The rats in the EA+VEGF group were also injected with 10 µL of VEGF165 (0.025 µg/µL) into the lateral ventricle after the first session of EA. Five rats in each group were sacrificed after obtaining a neurological function score (mNSS) at day 0 (1 d after modeling, before EA intervention), day 7 and day 14, respectively. Nissl staining was used to observe the histomorphology of cerebral infarction areas. Immunohistochemistry was used to CM(155mm]detect GRP78 activity in the ischemic brain tissues. Real-time fluorescence quantitative PCR (real-time PCR) was used to detect the expressions of caspase12, caspase3 and GRP78 mRNA in the ischemic brain tissues. RESULTS: Compared with the sham-operation group, rats in the model group had higher mNSS scores ( P<0.05), showed signs of cerebral infarction (with reduced numbers of and disordered Nissl bodies and unclear structure), increased GRP78 immunopositive cells, increased expression of GRP78 mRNA ( P<0.05), and increased expressions of caspase12 and caspase3 mRNA ( P<0.05). Compared with the model group, EA and EA+VEGF decreased mNSS scores at day 7 and 14 ( P<0.05), showing alleviated signs of cerebral infarction, increased GRP78 immunopositive cells ( P<0.05), increased GRP78 mRNA expression ( P<0.05), and decreased caspase12 and caspase3 mRNA expressions ( P<0.05). The most obvious changes were found in the EA+VEGF group ( P<0.05). No significant changes were observed in the sham-operation group over time ( P<0.05). In comparison, mNSS scores, the signs of cerebral infarction, and the expressions of caspase12 and caspase3 decreased over time in the other groups ( P<0.05), accompanied with increased GRP78 immunopositive cells and the expression of GRP78 gene ( P<0.05). CONCLUSION: Electroacupuncture and intracerebral injection of VEGF promote tissue repair of rats with cerebral ischemic injury, possibly through down-regulating the expressions of caspase12 and caspase3 genes and up-regulating the expression of GRP78 gene. The effect of electroacupuncture in combination with intracerebral injection of VEGF is superior to that of the single use of electroacupuncture.

Global properties of vector-host disease models with time delays.

Since there exist extrinsic and intrinsic incubation periods of pathogens in the feedback interactions between the vectors and hosts, it is necessary to consider the incubation delays in vector-host disease transmission dynamics. In this paper, we propose vector-host disease models with two time delays, one describing the incubation period in the vector population and another representing the incubation period in the host population. Both distributed and discrete delays are used. By constructing suitable Liapunov functions, we obtain sufficient conditions for the global stability of the endemic equilibria of these models. The analytic results reveal that the global dynamics of such vector-host disease models with time delays are completely determined by the basic reproduction number. Some specific cases with discrete delay are studied and the corresponding results are improved.

[Effect of Electro-acupuncture on the Expression of Slit2 and HSPGs in Rats with Focal Cerebral Infarction].

OBJECTIVE: To determine the expression of Slit2 and heparan sulfate proteoglycans (HSPGs) in cortex of rats with focal cerebral infarction and the effect of acupuncture (EA) on the expression of Slit2 and HSPGs. METHODS: 40 male Sprague Dawley (SD) rats were equally randomized into four groups: Control group, model group, non-acupoint EA group, and EA group. Thread-tying method was used in the model group, non-acupoint EA group and EA group to clog arteries and then open up after 1.5 h. Morphology changes of tissues around the infarction area were observed 14 days later by Nissl staining. The expressions of Slit2 and HSPGs in the ischemic brain tissues were detected by indirect immunofluorescence staining (IF) and Western blot (WB). RESULTS: Modified neurologicalseverity scores (mNSS) showed zero in the control group, lower than the score of EA group. The EA group had lower mNSS score than the non-acupoint group. The highest mNSS score appeared in the model group. Paired comparisons showed statistical differences (P < 0.01). Nissl's staining showed that EA group had increased Nissl bodies in alignment; non-acupoint EA group had increased and disordered Nissl bodies; model group had decreased and disordered Nissl bodies with edema in the brain. IF and WB showed that non-acupoint EA group had higher levels of Slit2 and HSPGs than model group (P < 0.05); EA group had higher levels of Slit2 and HSPGs than non-acupoint EA group (P < 0.05); model group had higher levels of Slit2 and HSPGs than control group (P < 0.05). The results of Nissl' s staining, IF and WB were consistent. CONCLUSION: EA can enhance the expressions of Slit2 and HSPGs. This may be one of the mechanisms of EA promoting recovery of neural functions after cerebral infarction.

[Effect of Electroacupuncture with Near-to-bone Needling to Chondrocyte Apoptosis and Proliferation on the Knee Osteoarthritis Model].

OBJECTIVES: To study the effect of electroacupuncture with near-to-bone needling on the proliferation and apoptosis of knee osteoarthritis. METHODS: Totally 40 New Zealand rabbits were randomly divided into two groups, the normal group (A, n =10) and the modeling group (n =30). Hulth-Telhag method was applied 30 rabbits to establish rabbit left knee osteoarthritis model, and evaluated by X-ray after 6 weeks. Then divided the 30 modeling rabbits into electroacupuncture with near-to-bone needling group (C,n =10), electroacupuncture with ordinary needling group (D,n =10) and model group (B ,n =10) randomly. Four treatments (5 d as one course of treatment) were applied to C group and D group, respectively. At the end of the experiment, modeling situation was detected by X-ray, pathological changes of cartilage was detected by HE staining ,and apoptosis of chondrocytes was checked by TUNEL staining; mRNA expressions of Caspase-3 and proliferating cell unclear antigen (PCNA) in chondrocytes were detected by RT-PCR; protein expressions of Caspase-3, PCNA, Bcl-2 and Bax in chondrocytes were detected by Western blot. RESULTS: HE stain demonstrated that the chondrocytes of A group,C group and D group were arranged in a more regular and smooth surface when compared with B group. The expressions of Bcl-2 and PCNA in C and D groups were higher than those of B group, but the expression of Caspase-3 and Bax were lower, when evaluated with Western blot;increased expressions of PCNA and Bcl-2 but decreased expressions of Caspase-3 and Bax in C group were also noticed when compared with D group. RT-PCR results demonstrated that when compared with B group, the expression of PCNA mRNA were increased but Caspase-3 mRNA were decreased in C and D groups; compared with D group, the expression of PCNA mRNA were increased but Caspase-3 mRNA were decreased in C group. CONCLUSIONS: The electroacupuncture with near-to-bone needling method is superior to the electroacupuncture with ordinary needling in decreasing apoptosis and promoting proliferation of chondrocytes.

Identification of a key motif that determines the differential surface levels of RET and TrkB tyrosine kinase receptors and controls depolarization enhanced RET surface insertion.

The RET tyrosine kinase receptor plays an important role in the development and maintenance of the nervous system. Although the ligand-induced RET signaling pathway has been well described, little is known about the regulation of RET surface expression, which is integral to the cell ability to control the response to ligand stimuli. We found that in dorsal root ganglion (DRG) neurons, which co-express RET and TrkB, the receptor surface levels of RET are significantly higher than that of TrkB. Using a sequence substitution strategy, we identified a key motif (Box1), which is necessary and sufficient for the differential RET and TrkB surface levels. Furthermore, pharmacological and mutagenesis assays revealed that protein kinase C (PKC) and high K(+) depolarization increase RET surface levels through phosphorylation of the Thr(675) residue in the Box1 motif. Finally, we found that the phosphorylation status of the Thr(675) residue influences RET mediated response to GDNF stimulation. In all, these findings provide a novel mechanism for the modulation of RET surface expression.

Analysis of an environmental epidemic model based on voluntary vaccination policy.

With the worsening of the environment and the increasing international trade, indirect transmission from exposure to contaminants in the surrounding environment has become an overlooked mode of transmission. This paper proposes a new game-theoretic model considering voluntary vaccination against imperfection and the unique integration of human-to-human and virus-to-human transmission routes. Based on the individual-based risk assessment update rule (IB-RA), the strategy-based risk assessment update rule (SB-RA), and the direct commitment update rule (DC), the different effects of individuals' behaviors on disease prevalence are analyzed. To find the effect of indirect transmission on epidemic transmission, we compare our model with the traditional SVIR model. Finally, it can be seen that indirect transmission mechanisms will aggravate the spread of epidemics.

[Electroacupuncture intervention improves lipid metabolism and promotes browning of white adipose tissue by activating AMPK/Sirt1 pathway and up-regulating Nrg4 content in middle-aged and aged obese rats].

OBJECTIVE: To investigate the mechanism of electroacupuncture （EA） in promoting the browning of white adipose tissue in middle-aged and aged obese rats induced by high fat by regulating AMP-activated protein kinase （AMPK） /silence-information regulatory factor 1 （Sirt1） pathway and neuregulin 4 （Nrg4）. METHODS: Twenty-four male SD rats were randomized into blank control, model and EA groups （n=8 per group）. The obesity model was established by feeding the rats with high-fat diet for 6 weeks. For the EA group, EA （2 Hz/15 Hz, 1.5 mA） was applied to "Guanyuan" （CV4） and bilateral "Shenshu" （BL23）, "Fenglong" （ST40） and "Tianshu" （ST25） for 20 min, once a day, 5 days a week for 6 weeks. Rats of the blank control and model groups were also restrained for 20 min. The body mass and food intake were measured every week, and the Lee's index, epididymal fat, perirenal fat and brown adipose tissue were weighed. The contents of serum total cholesterol （TC）, triglyceride （TG）, high density lipoprotein cholesterol （HDL-C）, low density lipoprotein cholesterol （LDL-C） and norepinephrine （NE） were determined by ELISA. H.E. staining was used to observe the morphological changes of white and brown adipose tissue. The mRNA expression levels of mitochondrial uncoupling protein 1 （UCP1）, peroxisome proliferator activated receptor γ co-activator 1α （PGC-1α）, PR-domain protein 16 （PRDM16）, peroxisome proliferator activated receptor γ （PPARγ） and Nrg4 in the adipose tissue were detected by quantitative real time PCR, and the protein expression levels of Nrg4, AMPKα, Sirt1 and interleukin-6 （IL-6） in the white and brown adipose tissue were detected by Western blot. RESULTS: Compared with the blank control group, the body mass, food intake, the Lee's index, epididymal fat and perirenal fat mass, and serum TG, TC and LDL-C contents and the expression level of IL-6 protein were significantly increased （P<0.01, P<0.05, P<0.001）, and the brown adipose mass, serum HDL-C and NE contents, the expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and the protein expression levels of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were significantly decreased in the model group （P<0.01, P<0.05）. After EA intervention, the increased levels of body mass, food intake, Lee's index, epididymal fat and perirenal fat mass, serum TG, TC and LDL-C contents, and the expression of IL-6 protein, and the decreased levels of brown adipose mass, serum HDL-C and NE contents, expression levels of UCP1, PGC-1α, PRDM16, PPARγ and Nrg4 mRNAs, and those of AMPKα, Sirt1 and Nrg4 proteins in both white and brown adipose tissues were apparently reversed（P<0.05, P<0.01, P<0.001）. H.E. staining showed an increase of the volume and content of intracellular vacuoles of both white and brown adipose tissues, disordered arrangement of cells with vague boundary in the model group, which was relatively milder including a decrease of volume and content of vacuoles of both white and brown adipose, neat arrangement of cells with clear boundary. CONCLUSION: EA intervention can improve lipid metabolism and promote white adipose tissue browning in middle-aged and aged obese rats, which is possibly associated with its functions in activating AMPK/Sirt1 signaling pathway and up-regulating the level of Nrg4.

[Fibrin glue-assisted lamellar keratoplasty for the treatment of corneal ulcer].

OBJECTIVE: To investigated the feasibility of fibrin glue-assisted sutureless lamellar keratoplasty (LK) in patients with corneal ulcer. METHODS: Noncomparative interventional case series. In 21 eyes of 21 patients with terriens marginal degeneration (TMD) or corneal ulcer after trauma, sutureless lamellar keratoplasty (LK) using fibrin glue was performed. Postoperatively, the graft status, graft clarity, the visual prognosis and postoperative complications were recorded. RESULTS: There were 16 cases suffered with Terriens marginal degeneration (TMD) and 5 cases with corneal ulcer after foreign body removed. All the corneal ulcers were successfully healed after the procedure, the operative complications included corneal perforation during operation in 1 case, recurrence of TMD in 1 case and graft opacity in 2 cases. There was no graft dislocation, hydrops between graft and recipient interface, secondary infection and graft rejection after surgery, 19 cases had improved best-corrected visual acuity (BCVA) while 2 cases had poor vision acuity due to graft opacity (t = -2.587, P = 0.018). CONCLUSION: Fibrin glue-assisted sutureless LK showed excellent graft stability in all patients.

Using an age-structured COVID-19 epidemic model and data to model virulence evolution in Wuhan, China.

COVID-19 is a disease caused by infection with the virus 2019-nCoV, a single-stranded RNA virus. During the infection and transmission processes, the virus evolves and mutates rapidly, though the disease has been quickly controlled in Wuhan by 'Fangcang' hospitals. To model the virulence evolution, in this paper, we formulate a new age structured epidemic model. Under the tradeoff hypothesis, two special scenarios are used to study the virulence evolution by theoretical analysis and numerical simulations. Results show that, before 'Fangcang' hospitals, two scenarios are both consistent with the data. After 'Fangcang' hospitals, Scenario I rather than Scenario II is consistent with the data. It is concluded that the transmission pattern of COVID-19 in Wuhan obey Scenario I rather than Scenario II. Theoretical analysis show that, in Scenario I, shortening the value of L (diagnosis period) can result in an enormous selective pressure on the evolution of 2019-nCoV.

[Effect of mild moxibustion on vascular aging in senescence rats based on local MCP-1/MMP-2/TGF-ß1 pro-inflammatory signal loop].

OBJECTIVE: To observe the effect of mild moxibustion on monocyte chemotaxis protein 1 (MCP-1)/matrix metalloproteinase 2 (MMP-2)/transforming growth factor ß1 (TGF-ß1) pro-inflammatory signal loop in senile rats, so as to explore its mechanisms underlying improving vascular aging (VA). METHODS: Twenty-four male VA SD rats were randomized into senium (VA) control, medication and moxibustion groups, and other 8 young SD rats (aged 2 months) were used as the young control group. The VA model was established by intraperitoneal injection of D-galactose (300 mg·kg-1·d-1) once daily for 4 weeks, and verified by serum total testosterone (TT) and free testosterone (FT) levels. For rats of the moxibustion group, mild moxibustion was applied to bilateral "Shenshu"(BL23) and "Guanyuan"(CV4) for 20 min, once a day, 5 days a week for 8 weeks. Rats of the medication group were treated by intraperitoneal injection of testosterone propionate (7 mg·kg-1·[3 d]-1) once daily for 8 weeks except weekends, and rats of the senium control and young control groups treated by intraperitoneal injection of the same dose of normal saline, once daily for 8 weeks except weekends. The duration of exhausted swimming (DES) before and after the treatment was recorded. H.E. staining and Masson staining were used to observe histopathological changes and collagen fiber content of the thoracic aortic tissue, respectively. The contents of serum TT, FT and angiotensin 2 (Ang Ⅱ) were determined by ELISA. The immunoactivity of aortic MCP-1 was detected by immunohistochemistry, and the expression levels of aortic MCP-1, MMP-2 and TGF-ß1 were detected by Western blot. RESULTS: Compared with the young control group, the levels of DES, serum TT and FT contents were significantly decreased (P<0.01), while those of serum AngⅡ and collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression considerably increased in the senium control group (P<0.01). After the interventions, the decreased levels of DES, serum TT and FT contents and the increased levels of serum AngⅡ, collagen fiber contents, aortic MCP-1 immunoactivity and MMP-2, TGF-ß1 and MCP-1 protein expression were reversed in both medication and moxibustion groups (P<0.01, P<0.05). The therapeutic effect of mild moxibustion was significantly superior to that of medication in down-regulating the aortic collagen fiber, serum AngⅡ contents and MCP-1 immunoactivity and protein expression (P<0.05). H.E. staining showed thickened endometrium and disordered arrangement of vascular smooth muscles of the aorta in the senium group, and thinner endometrium and regular and ordered arrangement of aortic vascular smooth muscles in both moxibustion and medication groups. CONCLUSION: Mild moxibustion may improve vascular aging in senescence rats, which is possibly by suppressing vascular MCP-1/MMP-2/TGF-ß1 pro-inflammatory signal loop.

Mechanism underlying activity-dependent insertion of TrkB into the neuronal surface.

Activity-dependent insertion of tyrosine kinase receptor type 2 (TrkB receptor) into the plasma membrane can explain, in part, the preferential effect of brain-derived neurotrophic factor (BDNF) on active neurons; however, the detailed cellular and molecular mechanisms underlying this process are still unclear. In our study, we developed a fluorescence ratiometric assay for surface TrkB receptors to investigate the mechanisms of recruitment of TrkB to the plasma membrane following chemical long-term potentiation (cLTP) induction. We found that, in hippocampal neurons, the effect of cLTP-induced TrkB surface-recruitment occurred predominantly on neurites with rapid kinetics (t(1/2) of approximately 2.3 minutes) and was dependent on an intact cytoskeleton structure. Mutagenesis studies revealed that the juxtamembrane domain of TrkB is necessary and sufficient for its activity-dependent insertion into the plasma membrane. Moreover, we found that the phosphorylation of TrkB receptor at the Ser478 site by cyclin-dependent kinase 5 (Cdk5) is essential for cLTP-induced TrkB insertion into the neuronal surface. Finally, the degree of cLTP-induced TrkB surface-recruitment is higher in postsynaptic regions, which provides a potential mechanism for rapid enhancement of postsynaptic sensitivity to incoming BDNF signaling. Our studies provide new insights regarding neuronal activity-dependent surface delivery of TrkB receptor, which will advance our understanding of the modulatory role of TrkB in synaptic plasticity.

[Non-Descemet stripping endothelial keratoplasty for treating iridocorneal endothelial syndrome in phakic eyes].

OBJECTIVE: To investigate the feasibility and clinical effects of non-Descemet stripping endothelial keratoplasty (nDSEK) on treating iridocorneal endothelial (ICE) syndrome in phakic eyes. METHODS: Retrospective noncomparative interventional case series. 7 patients (7 eyes) with ICE syndrome at the Affiliated Xiamen Eye Center of Xiamen University from January 2008 to December 2009 underwent non-Descemet stripping endothelial keratoplasty. All patients were followed up for 3 - 12 months, pre- and postoperative best corrected visual acuity (BCVA) were compared, the adherence of the donor disc to the recipient endothelium and postoperative donor disc dislocation were monitored during the follow-up period. Graft clearance and endothelial cell density (ECD) were observed, too. RESULTS: After nDSEK, no primary graft failures dislocation and decentered graft occurred during the follow-up period. Study group intraoperative complications included 3 case with elevated intraocular pressure 2 day postoperatively. Subepithelial haze, donor-recipient interface haze, and interface particles were observed in all measurable cases by in vivo laser confocal microscopy. Anterior segment optical coherence tomography and ultrasound biomicroscopy showed the adherence of the donor disc to the recipient endothelium and peripheral anterior synechiae were separated. All corneas remained clear during the follow-up. 6 patients had improved BCVA while 1 patient had the same BCVA. The reason for poor VA was optic atrophy due to glaucoma. Postoperative mean EDC was (2176.6 ± 267.6) cells/mm(2). CONCLUSIONS: nDSEK for iridocorneal endothelial (ICE) syndrome is feasible, technically easy, safe and effective. It can be one of the surgical treatment option for bullous keratopathy.

Modeling and optimal control analysis of COVID-19: Case studies from Italy and Spain.

Coronavirus disease 2019 (COVID-19) is a viral disease which is declared as a pandemic by WHO. This disease is posing a global threat, and almost every country in the world is now affected by this disease. Currently, there is no vaccine for this disease, and because of this, containing COVID-19 is not an easy task. It is noticed that elderly people got severely affected by this disease specially in Europe. In the present paper, we propose and analyze a mathematical model for COVID-19 virus transmission by dividing whole population in old and young groups. We find disease-free equilibrium and the basic reproduction number (R 0). We estimate the parameter corresponding to rate of transmission and rate of detection of COVID-19 using real data from Italy and Spain by least square method. We also perform sensitivity analysis to identify the key parameters which influence the basic reproduction number and hence regulate the transmission dynamics of COVID-19. Finally, we extend our proposed model to optimal control problem to explore the best cost-effective and time-dependent control strategies that can reduce the number of infectives in a specified interval of time.

Coinfection dynamics of heroin transmission and HIV infection in a single population.

We propose a model of a joint spread of heroin use and HIV infection. The unique disease-free equilibrium always exists and it is stable if the basic reproduction numbers of heroin use and HIV infection are both less than 1. The semi-trivial equilibrium of HIV infection (heroin use) exists if the basic reproduction number of HIV infection (heroin use) is larger than 1 and it is locally stable if and only if the invasion number of heroin use (HIV infection) is less than 1. When both semi-trivial equilibria lose their stability, a coexistence equilibrium occurs, which may not be unique. We compare the model to US data on heroin use and HIV transmission. We conclude that the two diseases in the US are in a coexistence regime. Elasticities of the invasion numbers suggest two foci for control measures: targeting the drug abuse epidemic and reducing HIV risk in drug-users.

[Detection of DNA damage of workers occupationally exposed to lead with flow cytometer assay].

OBJECTIVE: To study DNA damage of workers occupationally exposed to lead with flow cytometer assay. METHODS: The lymphocytes were obtained from 41 workers occupationally exposed to lead (comparable group) and another 50 from control group. Flow cytometer (FCM) assay was used to detect DNA damage. RESULTS: DNA damage rate and geometric mean fluorescence intensity in the comparable group were significantly higher than those in the control group (P<0.05). There were no significant differences in the DNA damage and geometric mean fluorescence intensity between different age groups (P>0.05). The differences in correlation analysis between blood lead, urine lead, delta-ALA and DNA damage rate were not significant (P>0.05). The correlation analysis showed no statistical significance between concentration of blood lead, urine lead, delta-ALA and geometric mean fluorescence intensity (P>0.05). There was positive correlations not only between the high concentration of blood lead, delta-ALA and damage rate of DNA, but also between the high concentration of blood lead and geometric mean fluorescence intensity. The coefficient r showed statistical significance (P<0.05). CONCLUSION: Occupational lead exposure can cause DNA damage. Gamma H2AX flow cytometer assay is a sensitive, objective and effective method for detection of DNA damage of peripheral blood lymphocytes.

[Electroacupuncture intervention increases testosterone level of aged rats by activating ERK/Nrf2/HO-1 signaling of Leydig cells].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on anti-oxidant function of Leydig cells in aged rats with low testosterone, so as to reveal its underlying mechanism of anti-aging of male reproduction. METHODS: Eighteen 20 months old SD rats were divided into aged control, medication and EA groups(n=6 in each group), and other 6 young male SD rats (2 months of age) were used as the youth control group. The rats of the youth and the aged control groups received subcutaneous injection of 0.9% normal saline (7 mg·kg-1·3 d-1) for 8 weeks, and those of the medication group received abdominal subcutaneous injection of Testosterone Propionate (7 mg·kg-1·3 d-1) for 8 weeks. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral "Shenshu" (BL23) and "Guanyuan" (CV4) for 15 min, once daily for 8 weeks except the weekends. The levels of serum total testosterone (TT) and free testosterone (FT) were determined by enzyme linked immune sorbent assay (ELISA), the immunoactivity of phosphorylated extracellular signal-regulated protein kinase (p-ERK) was detected by immunohistochemistry, and the expression levels of ERK, p-ERK, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in the testis tissues were determined by Western blot. RESULTS: Before and after the treatment, the levels of serum TT and FT in the aged control group were significantly lower than those of the youth control group (P<0.01). After the treatment, the serum TT and FT contents in the EA and medication groups were obviously higher than those in the aged control group (P<0.01). Compared with the youth control group, the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins were significantly down-regulated in the aged control group (P<0.01), while in comparison with the aged control group, the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins were significantly up-regulated in the medication and EA groups (P<0.01). The therapeutic effect of EA was notably superior to that of medication in up-regulating the immunoactivity of p-ERK, and the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins (P<0.01). No significant differences were found between the EA and medication groups in the increased levels of serum TT and FT after the intervention (P>0.05). CONCLUSION: EA intervention may increase testosterone level of the aged rats, which may be related to its effects in triggering ERK /Nrf2 /HO-1 signaling (anti-oxidative stress signal pathway) in the testis.

[Warm needle moxibustion, mild moxibustion and electroacupuncture interventions have an anti-aging effect possibly by regulating hypothalamus-pituitary-testis axis in aged male rats].

OBJECTIVE: To observe the effect of warm needle moxibustion (WNM), mild moxibustion and electroacupuncture (EA) of "Shenshu" (BL23) and "Guanyuan" (CV4) on serum gonadal hormone levels and related gene expression in the hypothalamus and testis in aged rats, so as to explore their underlying mechanisms in anti-aging. METHODS: Forty male SD male rats were randomly divided into aged control, WNM, mild moxibustion, EA and medication groups (n ＝ 8 in each group), and other 8 male young adult SD rats were used as the youth control group. WNM (acupuncture plus moxibustion), mild moxibustion or EA (2 Hz /100 Hz, 1 mA) was separately applied to CV4 and bilateral BL23 for 15 min, once daily, 5 days a week for 8 weeks. Rats of the medication group received subcutaneous injection of testosterone propionate injection (7 mg/kg), once every 3 days, and those of the aged and youth groups received the same dose of 0.9% NaCl solution for 8 weeks. The levels of serum testoste-rone (T) and gonadotropin-releasing hormone (GnRH) were measured by ELISA. The expression of Kisspeptin (KISS-1), G protein-coupled receptors 54 (GPR54), RFamide-related peptide-3 (RFRP-3) and GPR147 mRNAs in the hypothalamus and GnRH-R, GPR54 and GPR147 mRNAs in the testis tissues were detected by quantitative real-time PCR. RESULTS: The levels of serum T and GnRH, and the expressions of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 mRNAs in the testis in the aged rats were significantly lower than those in the youth rats (P<0.05), while the expression level of GPR147 mRNA of the testis tissue was obviously higher in the aged group than that in the youth group (P<0.05). Follo-wing the treatment, the lowered serum T and GnRH contents, the down-regulated expression levels of hypothalamic KISS-1, GPR54, RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs, as well as the up-regulated testicle GPR147 mRNA expression in the aged rats were reversed in the WNM, mild moxibustion and EA groups (not the medication group except serum T) in comparison with the aged control group (P<0.05). The therapeutic effects of WNM, mild moxibustion and EA were significantly superior to those of medication in up-regulating serum GnRH content and expression levels of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R, GPR54 and GPR147 mRNAs in the testis (P<0.05). No significant differences were found among the WNM, mild moxibustion and EA groups in serum T and GnRH contents, and the expression levels of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 and GPR147 mRNAs in the testis (P>0.05)．. CONCLUSION: The WNM, mild moxibustion and EA therapies have an anti-aging effect by increasing serum T and GnRH levels in aged rats, which is possibly related to their effects in up-regulating the expression of hypothalamic KISS-1, GPR54, RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs, as well as down-regulating testicle GPR147 mRNA expression. The therapeutic effect of WNM, mild moxibustion and EA is evidently superior to that of medication.

[DMF-induced adult hepatocyte apoptosis and its effects on expression of Bcl-2, Bax and Caspase-3].

OBJECTIVE: To investigate the apoptosis-inducing effect of DMF on the human liver cells (HL-7702) in vitro. METHODS: Liver cells were exposed to different concentrations of DMF (0, 50, 100, 150, 200 mmol/L) for 12 hours. Apoptotic rate, the expression of Bax, Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively. RESULTS: The increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group (P < 0.01), the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance (P > 0.05), as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control (P < 0.01). The new lands of procaspase-3 in 150, 200 mmol/L were observed, which demonstrated that there was active caspase-3. CONCLUSION: DMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.