The Fourier Transform Infrared Spectra of the Key Organs Derived from Kidney(Shen)-Yang Deficiency Syndrome Mice

Objective：To explore an approach to rapidly and accurately identify the compounds as biomarkers of Chinese medicine（CM）syndromes.Method：The Fourier transform infrared（FT-IR）spectrometry was applied to investigate the characteristic components of a mice model of Kidney（Shen）-yang deficiency syndrome（KDS）,and the remedial effect of a typical CM formula Shenqi Pill（肾气丸）.Thirty-six females and 18 males of Balb/c mice were randomly divided into KDS,Shenqi or control group.The females and males of the same group freely were mated for 96 h,and the males were taken out and only the female mice were raised.Females of the KDS group were threatened by a ferocious cat every other day for 14 d.After delivery,the KDS,or gestational threatened,offspring were raised at standard condition for 11 weeks.Then 10 male offspring were randomly selected,anaesthetized and their representative organs,i.e.testes,kidneys,lungs and feet were collected,for the FT-IR scan.Mice of the Shenqi group were intragastric administered Shenqi Pill;while mice in the KDS and control groups were given the same volume of saline.Results：The attenuated birth outcomes of the KDS group were displayed.The remarkable FT-IR differences of all organs between KDS mice and healthy control were mainly at 1,735-1,745 cm-1（indicating the increased levels of lipids）and at 1,640-1,647 cm-1and 1,539-1,544 cm-1（displaying the decreased proteins）.No statistic FT-IR difference between Shenqi and control mice was observed.Conclusion：In accordance with major traits of KDS,prenatal stress extensively impaired the building up of proteins and resulting in the excessive lipid storage,and FT-IR could effectively identify the biomarkers of KDS.

Shen(Kidney)-Tonifying Principle for Primary Osteoporosis:to Treat Both the Disease and the Chinese Medicine Syndrome

Primary osteoporosis （POP） is one of the most common diseases in the elderly people resulting in high risk of fracture and poor quality of life. In addition to the pathological changes in bone mass, most of the POP patients also suffer from Chinese medicine （CM） syndromes of Shen （Kidney） essence deficiency. Shen essences are highly related to bone. Shen essence deficiency plays an important part in the development of POP and a better diagnosis of POP could be made by combining CM syndromes with Western medicine risk factors. Treatments of POP should aim at both increasing the bone mass and relieving the syndromes of Shen essence deficiency. Clinical study confirmed that treating POP patients with Shen-tonifying herbs could increase the bone mass and relieve the CM syndromes of POP patients. Basic researches clarified the mechanism by which Shen-tonifying herbs increased bone mass in animal models. The mechanisms by which Shentonifying herbs relieve the CM syndromes are still in investigation.

Essence of"Shen(Kidney)Controlling Bones":Conceptual Analysis Based on Hypothalamic-Pituitary-Adrenal-Osteo-Related Cells Axis

As a traditional concept of Chinese medicine（CM）, the theory of ＂Shen（Kidney） controlling bones＂ has been gradually proven. And in modern allopathic medicine, the multiple mechanisms of bone growth, development and regeneration align with the theory. Shen deficiency as a pathological condition has a negative effect on the skeleton of body, specifically the disorder of bone homeostasis. Present studies indicate that Shen deficiency shares a common disorder characterized by dysfunction of hypothalamic-pituitary-adrenal（HPA） axis. HPA axis may be an important regulator of bone diseases with abnormal homeostasis. Therefore, we posit the existence of hypothalamic-pituitary-adrenal-osteo-related cells axis： cells that comprise bone tissue（osteo-related cells） are targets under the regulation of HPA axis in disorder of bone homeostasis. Chinese herbs for nourishing Shen have potential in the development of treatments for disorder of bone homeostasis.

Strategies for Activating Blood Circulation-Regulating Gan(Liver)-Tonifying Shen(Kidney) Sequential Therapy of Endometriosis-Associated Infertility

Endometriosis(EM) is a major cause of infertility and has a complex pathogenesis,which brings severe challenges in both clinical treatment and scientific research.Current clinical research focuses on the difficult problem of improving the pregnancy rate of EM patients.Our studies found that Chinese medicine has significant advantages in terms of improving the pregnancy rate of EM patients.

Liuwei Dihuang Pill(六味地黄丸)Treats Postmenopausal Osteoporosis with Shen(Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression

Objectives： To investigate the mechanism of Liuwei Dihuang Pill （六味地黄丸, LDP） in treating postmenopausal osteoporosis （PMOP） with Shen （Kidney） yin deficiency. Methods： In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group （Group A）, PMOP Shen-yang deficiency group （Group B）, PMOP without Shen deficiency group （Group C）, and control group （Group N）. Real-time polymerase chain reaction （RT-PCR） and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 （CLCF1）, ankyrin repeat and SOCS box containing 1 （ASB1）, and proldneticin 2 （PROK2） genes and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. Results： The mRNA （P〈0.05） and protein （P〈0.01） expression levels of the CLCF1 gone in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gone were obviously up-regulated （P〈0.01）. After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated （both P〈0.01）, and the average bone density of the top femur had significantly increased （P〈0.05）. In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3. Conclusions： The CLCF1 gone is an important gone associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gone expression and activation of the JAK/STAT signaling pathway.

Effect of Electroacupuncture on Spermatogenesis in Rats with Oligozoospermia of Insufficiency of Shen(Kidney) Essence Syndrome

Objective: To assess the effect of electroacupuncture(EA) on expression of cytoskeletal proteins from Sertoli cells(SCs) and spermatogenesis in rats with oligozoospermia of insufficiency of Shen(Kidney)essence syndrome(OIKES).Methods: Twenty healthy male Sprague-Dawley rats were randomly assigned to four groups using a random number table: control,tripterygium glycosides(TG) treatment,sham and EA groups(n=5 in each group).A rat model of OIKES was established by oral gavage with TG.The EA group was treated with TG and received EA at Shenshu(BL 23) and Zusanli(ST 36) acupoints for 20 min,once daily for 30 days,while the sham group received EA at identical acupoints with skin penetration without stimulation.After 30 days,the ?nal body weight and coef?cients for the testis and epididymis were calculated and sperm parameters were measured.Immunohistochemical analyses were performed to detect expression of vimentin and α-tubulin in SCs and proliferating cell nuclear antigen(PCNA) immunoreactivity in germ cells.Apoptosis in germ cells was quanti?ed by the transferase biotin-dUTP nick end labeling assay.Results: Compared with the control group,the final body weight and testis/epididymis coefficients of rats in the TG-treated group were not significantly different,but the sperm count and motility were lower(P<0.05).Expressions of vimentin and α-tubulin were also signi?cantly weaker(P<0.01).The PCNA immunoreactivity of germ cells was decreased(P=0.059),whereas the apoptotic index of germ cells was increased signi?cantly(P<0.01).In contrast,EA at BL 23 and ST 36 acupoints signi?cantly improved the ?nal body weight as well as the sperm count,concentration and motility(P<0.01 or P<0.05).EA increased expression of vimentin and α-tubulin in SCs markedly,and signi?cantly enhanced PCNA immunoreactivity with decreased apoptosis in germ cells(P<0.01 or P<0.05).Conclusions: EA at BL 23 and ST 36 acupoints has protective effects on spermatogenesis in rats with OIKES.This effect seems to be achieved by attenuating TG-induced disruption of cytoskeletal protein in SCs.

Shen Qi Wan attenuates renal interstitial fibrosis through upregulating AQP1

Renal interstitial fibrosis(RIF)is the crucial pathway in chronic kidney disease(CKD)leading to the end-stage renal failure.However,the underlying mechanism of Shen Qi Wan(SQW)on RIF is not fully understood.In the current study,we investigated the role of Aquaporin 1(AQP1)in SQW on tubular epithelial-to-mesenchymal transition(EMT).A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo.Subsequently,the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown.The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine,increased the protein expression of E-cadherin and AQP1 expression,and decreased the expression of vimentin andα-smooth muscle actin(α-SMA).Similarly,treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells.The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1.AQP1 knockdown also increased the mRNA expression of vimentin andα-SMA,and decreased the expression of E-cadherin.The protein expression of vimentin increased,while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells.These results revealed that AQP1 knockdown promoted EMT.Furthermore,AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells.In sum,SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.

Efficacy and Safety of Guilingji Capsules(龟龄集胶囊)for Treating Mild-to-Moderate Cognitive Impairment:Study Protocol for A Randomized,Double-Blind,Positive-Controlled,Multicenter and Noninferiority Trial

Background:The incidence of cognitive impairment(CI)is gradually increasing,which has attracted more attention from medical researchers worldwide.Definitive mechanisms of pathogenesis remain elusive,and there are few medications that have been proven effective for CI.The utilization of Chinese herbal medicine has shown positive therapeutic effects for a broad spectrum of diseases,including CI.Objective:The purpose of this study is to evaluate the safety and efficacy of Guilingji Capsules(GLJC,龟龄集胶囊)in treating mild-to-moderate CI with Shen(Kidney)and marrow deficiency syndrome.Methods:This is a randomized,double-blind,positive-controlled,multicenter clinical trial with a noninferiority design that included 348 participants randomly divided into an experimental arm and an active comparator arm.Individuals in the experimental arm(174 cases)took 0.6 g of GLJC once a day and 19.2 mg of Gingko biloba extract mimetic 3 times a day.Individuals in the active comparator arm(174 cases)took 0.6 g of GLJC mimetic once a day and 19.2 mg of Gingko biloba extract in tablet form 3 times a day.The intervention period included two sessions over 24 weeks.The primary outcome be the effectiveness of GLJC on cognitive improvement after 24 weeks of treatment,which was defined as an increase in the Mini Mental State Examination(MMSE)and Montreal Cognitive Assessment(MoCA)Scale.The secondary outcomes were improvement in independence,daily living ability,and Chinese medicine(CM)syndrome,which were measured with the Alzheimer’s disease Rating Scale-Cognitive Project(ADAS-Cog),Clinical Dementia Rating(CDR)Total Score,Activities of Daily Living(ADL)Total Score and the Chinese Medicine Symptom Scale(CM-SS),respectively.Serum acetylcholine,acetylcholinesterase,bax and bcl-2 were monitored to explore the mechanism of GLJC on CI.In addition,safety measures,including vital signs,electrocardiography,laboratory indicators(full blood count,kidney and liver function tests,routine urine test and routine stool test)and adverse events,were also recorded.Discussion:The purpose of this trial is to evaluate the efficacy and safety of GLJC in patients with mild-to-moderate CI with kidney and marrow deficiency syndrome.If successful,the results would provide a viable treatment for patients with mild-to-moderate CI.

Microenvironment of Liver Regeneration in Liver Cancer

The occurrence and development of liver cancer are essentially the most serious outcomes of uncontrolled liver regeneration. The progression of liver cancer is inevitably related to the abnormal microenvironment of liver regeneration. The deterioration observed in the microenvironment of liver regeneration is a necessary condition for the occurrence, development and metastasis of cancer. Therefore, the use of a technique to prevent and treat liver cancer via changes in the microenvironment of liver regeneration is a novel strategy. This strategy would be an effective way to delay, prevent or even reverse cancer occurrence, development and metastasis through an improvement in the liver regeneration microenvironment along with the integrated regulation of multiple components, targets, levels, channels and time sequences. In addition, the treatment of ＂tonifying Shen（Kidney） to regulate liver regeneration and repair by affecting stem cells and their microenvironment＂ can regulate ＂the dynamic imbalance between the normal liver regeneration and the abnormal liver regeneration＂; this would improve the microenvironment of liver regeneration, which is also a mechanism by which liver cancer may be prevented or treated.